(MT 6326) IMMUNOSERO LEC COMPRE by 3DMT PDF
(MT 6326) IMMUNOSERO LEC COMPRE by 3DMT PDF
(MT 6326) IMMUNOSERO LEC COMPRE by 3DMT PDF
1
SOURCES: PPT, A ch Cla LEGENDS: , lec e, b k
he ir s for pro ec ion ia
NI 1: IN ROD C ION O I AND HI ORICAL inhala ion
CONCEP E J
Smallpo accina ion (adminis ered
PRE- E 1798
in ram sc larl )
S ccessf ll pre en ed
1. Who disco ered he HLA? Jean Da sse
infec ion i h smallpo b
2. Variola ion is a form of imm ni a ion b inhaling dried
injec ing co po
po ders deri ed from smallpo lesions
3. Who in rod ced accina ion? Ed ard Jenner 1862 H E Phagoc osis
4. Who disco ered li e a en a ed accines for rabies? Lo is
L P
Pas e r
Li e a en a ed, chicken cholera and
5. The scien is ho firs e plained cell lar media ed imm ni
an hra accine
hro gh phagoc osis as Elie Me chnikoff.
Fa he of Imm nolog
Disco ered he firs a en a ed
IMM NOLOG & EROLOG accine
A e a ed = Red ced and
IMM NOLOG 1862
enforced effec (less ir len ) b
Resis ance o disease (infec io s disease)
applica ion of hea , aging or
S d of a hos s reac ions hen foreign s bs ances are
chemical means o lessen
in rod ced in o he bod
pa hogenici of microorganisms
Branch of biolog ha co ers he s d of imm ne s s ems
Obser ed ha older bac erial
in all organisms
c l res o ld no ca se
Consis s of:
disease in chickens
S d of molec les (an igens/an ibodies), organs
( ransplan a ion/compa ibili ), s s ems L P
responsible for he recogni ion and disposal of Therape ic accina ion
1885
foreign ma erial (RBCs & macrophages) Firs repor of li e a en a ed accine
Ho bod componen s respond and in erac for rabies
Desirable and ndesirable conseq ences of
E M
imm ne in erac ions
Cell la he fi i hro gh
Wa s in hich he imm ne s s em can be
phagoc osis
ad an ageo sl manip la ed o pro ec agains or
Cond c ed sing s arfish
rea disease
lar ae in hich i eng lfed
EROLOG
foreign ma erials
Scien ific s d of ser m and o her bod fl ids
1883-1905 Foreign objec s become
S d of he fl id componen s in he blood
s rro nded b mo ile
ameboid-like cells a emp ing
HI ORICAL BACKGRO ND o des ro pene ra ing objec s
Imm ni o disease as based
recorded ha indi id als
on ac ion of sca enger cells
ho had pre io sl con rac ed he
(inna e hos defense)
430 BC disease reco ered
He recogni ed heir imm ne s a s E B & K
H al he fi i or H moral
Chinese prac iced a form of
response proposed
imm ni a ion b inhaling dried po ders
AKA an ibod media ed
1000 AD deri ed from smallpo lesions
1890 imm ni
(1500s ( )
Aspec of imm ni hich is
acc di g Lesions ca sed b Variola ir s
media ed b he secre ed
S e e ) Va i la i - ma erials from
an ibodies
smallpo lesions/p s les are
Responsible cells B cells
gi en o people ho ne er had
P D PO - E
1986
Monoclonal Hepa i is B accine
R. M
Th1 ers s Th2 model of T-helper cell
f nc ion
1986 Th1 = s im la es he cell lar
imm ne response (T-cell
Th2 = s im la es he h moral
imm ne response (B-cell)
A e :B-D-A-E-C
J H nd B
a B
1996-1998
Iden ifica ion of Toll-like recep or
MMAR FOR RE IE
Yea Scien i Di co e
1000 AD (1500s) Chinese Variola ion (inhaling dried po er from smallpo lesions)
1890 Emil Von Behring & Shibasab ro Ki asa o H moral heor of imm ni
1902 Charles Por ier & Pa l Riche Immedia e-h persensi i i anaph la is
1938 John Richardson Marrack H po hesis of an igen-an ibod binding (imm ne comple )
1964-1968 Henr Claman T-cell & B-cell coopera ion in imm ne coopera ion
1972 Gerald M. Edelman & Rodne R. Por er Iden ifica ion of an ibod molec le
1975 George Kohler & R. Cesar Mils ein Firs monoclonal an ibodies
1985-1987 James P. Allison Iden ifica ion of genes for T-cell recep or
1996-1998 J les Hoffman & Br ce B ler Iden ifica ion of Toll-like recep or
S ce : PPT, S c Ca e , B a d e e e e ce
P ima f ac i i i n f
UNIT 2: HUMAN IMMUNE SYSTEM imm n c m e enc . In he ima
l m h id gan , nc mmi ed cell a e
an f med in l m h id cell (i.e.
PRE-TEST h m and b ne ma )
1. A componen of he inna e defen e em e ing a he Whe e ma a i n f B & T
e e nal p o ec i e hield of he bod again coloni ing l m h c e ake lace
mic obe Seconda e e a a " aining
a. In ac Skin g nd" f he c mmi ed l m h id cell
b. M c memb ane he ein he ld mee he an igen/
c. Phag c e
d. N mal fl a gani m f eign agen .
2. La ge l mphoid o gan Si e f final ma a i n and
a. A endi diffe en ia i n
b. S leen P ide l ca i n he e c n ac
c. Th m
i h f eign an igen can cc
d. L m h N de
3. I i bo h a l mphoid and endoc ine o gan impo an in 2 l m h id gan - T n il ,
he de elopmen of f nc ional T-l mphoc e l m h n de , leen and he
a. B ne Ma a cia ed i e (i.e. m c al
b. S leen and c ane a cia ed
c. Th m
d. L m h N de l m h id i e )
4. (T/F) The e i a g ad al dec ea e in he i e and Ne k f cell , i e , and gan ha ide he b d
ec e o abili ie of he h m a one age mechani m e i infec i n and di ea e
5. (T/F) The l mphoc e a e he main le koc e C mm nica e, c llab a e, and c llec i el k
e pon ible fo he f nc ion of he L mpha ic em ge he
6. T-l mphoc e a e in ol ed in ha pe of imm ne
e pon e? Rec gni e, ne ali e and de a h gen
a. Cell media ed imm ne e n e Iden if and de cance cell
b. H m al imm ne e n e Rem al f n cell and i e damaged b
c. B h Cell media ed and H m al imm ne e n e a ma di ea e
d. Inna e imm ne e n e
Sea ch f f eign/n n elf an igen ha d n
7. Diffe en ia ed fo m of B-l mphoc e
a. Ma cell bel ng he b d & de hem
b. Pla ma cell Main ain eillance e a ea ance f ne
c. Mac hage f eign an igen m cell & de hem
d. L m h bla hile lea ing n mal/ elf an igen n heal h cell
8. Which of he follo ing cell a e capable of ecogni ing
fo eign an igen nha med
a. B-cell M n a c dina ed imm ne e n e again he
b. Dend i ic cell f eign agen
c. Lange han cell R f he d f Imm n l g
d. Mac hage
e. All f he ab e FUNCTIONS OF THE IMMUNE SYSTEM
9. (T/F) Ac i e ac i ed imm ni of confe ed o an
indi id al ho ha e eco e ed f om an illne o di ea e 1. Defen e again infec ion
10. (T/F) Pa i e imm ni p o ide lifelong p o ec ion f om Deficien imm ni e l in inc ea ed ce ibili
a di ea e infec i n (i.e. AIDS)
Al ec i e again he f eign b ance
IMMUNE SYSTEM ha a e n n-infec i
Vaccina i n b imm ne defen e and ec
Refe he L m ha ic em again infec i n
L m ha ic e el 2. Defen e again neopla m o mo
L m ha ic fl id (l m h) ha fl f m he P en ial f imm n he a f cance
l m ha ic e el c ming f m i e . 3. Inj e cell and ind ce pa hologic inflamma ion
C m nen f he l m h a e i e imila Imm ne e n e a e he ca e f alle gic,
i h bl d la ma. a imm ne, and he inflamma di ea e
L m h c e a e c ncen a ed in hi a ea
L m h id Ti e and O gan
Di ib ion Ci c la i n and Bl d Ti e
Ma gina ing l ci c la i n (ha e ecific
name
de ending n
hei i e
di ib i n)
T pe of F a ed S ained Facili a e
Phagoc o i Phag c i Phag c i
Chemo ac ic 1. Bac e ial 1. T an f ming G h
Fac o li eich ic acid Fac
2. C e ide - C5a 2. M n c e Chem ac ic
IMPORTANT RECEPTORS
Kille Inhibi o Recep o Kille Ac i a ion Recep o
1. KIR - Kille Inhibi ind cing a i he a ge
Rece cell ( i all -infec ed m
Reg la e he ac i n cell ):
f NK cell 1. CD16 - Fc ece f
P e en killing f elf IgG; f elec i e l i f
F m he i e he e DC enc n e ed he an igen, APC cell b de ec ing cell c a ed i h
ec nda l m h id gan e en i T-hel e cell he he he cell an ib die (im an in
In me ca e , APC ma e en hi he leen hen a ge ed b NK cell me h e en i i i
an igen a e e en ed in he bl d a e n mal/abn mal eac i n )
N mal cell ld 2. NKG2D
ha e MHC Cla I NOTE:
m lec le n he B h ece bind
face f he cell. If di ea ed and cance cell
ADDITIONAL NOTE:
If he n clea ed cell i n abn mal, ha an ab ndan MHC,
and n MICA and MICB, NK cell ill n de i
Thi i eg la ed b he KIR m lec le a ached n
he NK cell
NK cell a e al able ec e e IFN gamma m e he
ac i a i n f mac hage
ONTOGENY OF B CELLS
Ma e /diffe en ia e in he B ne Ma
B ne ma mal cell f m ecial niche
he e em cell & B-cell ec e ide (kee
B-cell ec l cali ed ecei e ignal f
diffe en ia i n)
In he BM, he e ill be he de el men f imma e,
imm n c m e en B-cell
Imma e cell a e in l ed in he an igen
Independen ha e f de el men
D n need he e ence f an igen de el
Once ma ed, hi ill be he ime he e he cell
ill ha e he an igen Dependen ha e (ha en in
he 2 l m h id i e )
Remembe : T-hel e cell ld nl in e ac i h cell ih Th ee Di inc Pha e f B-Cell De el men
MHC cla 2 m lec le ( e en in APC ) ia he TCR and 1. De el men f ma e imm n c m e en B cell
CD4 ma ke (An igen-Independen pha e)
Wi h he elea e f c kine (IL-2), n nde g e cl nal 2. Ac i a i n f B cell b an igen
e an i n (An igen-Dependen pha e)
Cl nal e an i n - dada 3. Diffe en ia i n f ac i a ed B cell in la ma cell
CD4+ T-hel e cell (effec al ead ) can ec e e hich d ce an ib die
IL ec i mac hage and ne hil . S me f
FUNCTIONAL COMPARTMENTS
ORGANIZATION OF THE IMMUNE SYSTEM
A. S em Cell / Gene a i e C m a men
B ne Ma
B. P ima / Cen al L m h id
A ea he e imm ne cell ac i e
imm n c m e enc THYMUS
Si e f an igen inde enden l m h ie i Bil bed gan and i l ca ed ab e he hea & bel he
Th m ma a i n f T-cell h id gland
B ne Ma P d ce HSC and Child en ha e la ge h m b a eg lde i
ma a i n f B and NK cell h ink (a i )
C. Sec nda / Pe i he al L m h id Si e f de el men f f nc i nal T-cell
Si e f an igen de enden diffe en ia i n Each l be i di ided in malle l b le
Enca la ed Co e : Imma e h m c e and h mic c ical
Unenca la ed (e.g. C ane IS, e i helial cell
MALT)
LYMPH NODE
NATURAL/INNATE/NATIVE IMMUNITY
Inhe en and al a e en in a heal h e n
I e n e i n n- ecific, gene al and anda di ed
I ece ( a e n ec gni i n ece = ha ec gni e
a h gen a cia ed memb ane ein and e en damage
a cia ed m lec la a e n ) a e iden ical and a e
e e ed b a a ic la e f cell
Pa e n ec gni i n ece can ec gni e
PAMPS een n he face f mic bial agen
Once he cell ec gni e he PAMPS, a
eac i n ill cc b i i ne- i e-fi -all
ince i i gene ic
Can al ec gni e DAMPS
C m nen a e ef med - e in lace a a e e c
I d e n eac again he n mal h
I mechani m f defen e e-e i he in a i n f he
f eign agen
N i e e i needed
Lack imm n l gic mem
IMMUNITY
MAJOR PILLARS OF IMMUNITY
Inna e Imm ne S em (Na al imm ni )
N n- ecific, in-b n imm ni em
Gene ic n eciali ed and eac he
agen nl a fe h
Fi e in h defen e again infec i n
If he inna e imm ni cann de he
f eign b d , ada i e imm ni ill en e
Adap i e Imm ne S em (Ac i ed imm ni )
C m le em ha m n a ecific imm ne
e n e a d f eign agen (an igen)
In l e he T can B l m h c e 2 P inci al e f eac i n f he inna e imm ne em:
Inflamma ion and An i i al defen e
LINES OF DEFENSE OF THE IMMUNE SYSTEM E e nal defen e mechani m
In e nal Defen e Mechani m
W a e a ce e e a a e e ed b
a e de e e a a e ec a e ?
SIMILARITIES
Di c imina i n
S me l ble fac
S me cell la fac
PRE-TEST Ca be a b a ce ( ec a , a f a ec e,
ga i ) ha ca be ec g i ed b he i e e
1. I i a b a ce ha ca eac i h a c e di g
ia ece f he ce (i.e. BCR, TCR a d TLR )
a ib d ha a a e i a i e e e
2. (T/F) The ec e he a ige i , he e i i C a ed BCR a d TCR, TLR a e c ide ed
i ge ic a ge e ic ( i ca de ec a f eig b d ) a d
3. The ______ i he a e i f a a ige ha a i c ide ed a a i a e i i
c e di g ece ca ec g i e A b a ce NOT ade b he b d (Non- elf); f eig
4. (T/F) F ag e ed e ide f a ce ed a ige h d
be c e ed MHC be ec g i ed b a T-ce ece b a ce
5. ________ a e che ica b a ce added acci e S b a ce ha eac i h a a ib d e i i ed T ce
i c ea e a i ge i c ea e he i e e e Ma be ab e e ke a i e e e
6. The h a MHC i ca ed he _______ S b a ce ha he I e S e ca ecifica
7. MHC C a ________ i e e ed i a c ea ed ce
ec g i e i h he he f Ag ece e e ed
8. (T/F) A ige a cia ed i h MHC C a II a e e e ed
CD4 T-ce ec e ed b h c e
9. (T/F) The T-ce ca ec g i e a a ige a de d i ic ce Epi ope/de e i a i e - a e i f he
e e i he ab e ce f he MHC ec e a ige ha he ece ca ec g i e
10. The ge e c di g f he MHC a e f d he h a TLR - ge e ic ec g i i
f ch e _______
BCR, TCR - ecific ec g i i
★ NOTE: NOT ALL A ige a e ab e i aea i e
*RECAP OF TYPES OF IMMUNITY AND ITS e e
FEATURES Bi di g a ha e b i a a i ae
e e
Inna e Imm ni (Na al imm ni )
N - ecifica a d ge e a i ed e e he ce ai
FUNCTIONAL TYPES OF ANTIGENS
a e i he a h ge = N i gic e
➢ I ge a ige i aei e e e
Adap i e Imm ni (Ac i ed imm ni )
➢ Ha e a a ige ha ca i a e e e;
Cha ac e i ed i h ecific ec g i i agai a h ge
i c e e a ige
L h c e (T a d B) - ai i e ce
➢ T e ge e f-a ige ha e e a ce (b d
Ha i ci a e ecifici , di e i , a d
i eac hi a ige )
e f he ada i e i i
T igge ed b A ige ca ab e f e ici i g a
i e e e IMMUNOGENS
A ige ca ab e f i a i g he I e Re e
INTRODUCTORY CONCEPTS No e: ALL IMMUNOGENS a e ANTIGENS; N a
a ige a e i ge
CONCEPT OF LIGAND-RECEPTOR BINDING
Mac ec e ca ab e f igge i g a ada i e i e
e e (i d ce f ai f a ib die e i i ed T
ce i i c ee h )
T igge i e e e f h c e
Ca be Th m -dependen Th m -independen
Biological A. Age ( ei ) he e i g
P ope ie of O de i di id a dec ea ed c j ga e i d ce a
an Indi id al e e a ige ic i ai i e e e
Ne a e d f e d EX. Pe ici i ,
i ge (i e e P ge e e, A i i
c e e de e ed) CHEMICAL
B. O e a Hea h Chemical Compo i ion
Ma i hed/fa ig ed/ e ed High MW a d C e
e ike S c a C e
cce f i e e e P ei a e be e ha
C. Ge e ic Va iabi i ca b h d a e , i id , a d
Li ked he MHC a d c eic acid
ece ge e a ed d i g T Comple i
a d B- h c e de e e 3D a e a d he be f
D. D e f I ge e i e
Do e - A ig ifica a i f Ro e of En and Do age
a i ge be e i ed Deg adabili and he Abili o be
i de f he ada i e P oce ed and P e en ed i h MHC
e e ake ace A ige - e e i g ce
I aei e fi deg ade he a ige h gh
e e ake ca e a ige ce i g bef e he
f a a f ca e e a ige ic e i e
a h ge hei face
The a ge he a i f he
FURTHER REQUIREMENTS FOR IMMUNOGENICITY
i ge , he highe he
i e e e i 1. Ge e ic edi ii f he i di id a ha i e ici
Ve a ge a ca e i i e e e
T & B-ce e a ce 2. B a d T ce e e i e f a i di id a
E. R e f I c a i 3. R e f ad i i a i
E a e :i a e , S bc a e
i ade a , bc a e , a I a e
De e i e hich ce O a ad i i e ed
ai i be i ed I a a a
TOLEROGENS
A ige ha i d ce e a ce
Se f a ige a e e ge ic
T e a ce - e i e e f he i e
e i e f-a ige
P e ada i e i eU e i e e
T e eg I ge
E e a e ge d e a di i i h
he e e a he ha e ha ce
EPITOPES
O a a i f he a ige i ec g i ed b i
c e di g ece
Thi i ca ed he Epi ope (a ige ic de e i a )
M ec a ha e c fig a i ha a e
ec g i ed b B T ce
Ma be e ea i g c ie diffe i g ecifici ie
Ca be linea & e en ial (c f ai a)
L a / e e ia = f d a i g e chai
C a a=f f di g f e i e
chai
T e f c fig a i i a a fac he he he
i be ec g i ed b he i e ce
Fig e 2-4: Cha ac e i ic of Hap en B cell
Rec g i e b h i ea a d c f ai ae i e
(A): Ha e bi d B -cell ecep o , b he ece e ai e e he face f a i ge
i de e de c i k: N ac i a i ( i ge ic) NOTE: A e i e ha i c f a i a (ca ab e
fc - i ki g face Ig ec e ) ca igge B
(B) Ha e /Ca ie c e c -i k ece : B ce ae ce ac i a i
ac i a ed a d begi d ci g a ib die (i ge ic) T cell
Rec g i e e i e a a a f he c e
(C) A h gh ha e ca bi d he a ib d bi di g i e f ed i h MHC he face f APC
(a ige ic), a f he a ib d -ha e c e e e ai E i e ca be ec g i ed b T ce if he
i de e de : The eac i ca be i a i ed a e deg aded i e ide a d e e ed ge he
i h MHC ec e
(D) Whe b d ca ie , he ha e c ib e he f ai
f a i ec ec ed a ice, hich i he ba i f he eci i a i
a d agg i a i eac i
CONSIDERATIONS:
I a a ige a fi d e i e : (1) i h he
a e a ea a ce (2) i h he diffe e
ecifici
Diffe e a ib d bi d he e i e
de e di g he ecifici
A a ge e : If he e i e a e i c e i i
each he , ca be he f i ea de ec i f
c e di g ece
U e a ed b hei c ei i e i i a = ead
c eac i cc = ece ca
ec g i e he a ige
E a e: F a Ag, b d g A&B
a ige i h bac e ia accha ide
OTHER TYPES OF ANTIGENS
He agg i ge Ag e e i RBC
Agg i ge I b e Ag i ed i agg i a i
P eci i i ge S b e Ag i ed i ha e cha ge
he eac i cc (PH: be i b e)
Vi a Pa ic e Ac ed e a i i g a ib die
Fig e 2-3: Linea . Confo ma ional Epi ope A e ge I d ce a e gie
A a h ac ge A e ge i ed i a a h ac ic
(A) Li ea e i e c i f e e ia a i acid a i ge eac i
e ide chai . The e a be e e a diffe e e e T i e ab ic d c (e i ); e a i ed b
chai a i- i
(B) C f ai a e i e e f he f di g f a e ide
chai / , a d e e ia a i acid a e b gh i c e
ADJUVANTS
i i
S b a ce ad i i e ed i h a i ge ha i c ea e
a d ha e he i e e e
REMEMBER! :) U a added acci e f a ie ha e a
be e i e e e he acci e
U a e ha ce he effec f he i ge b :
I c ea i g he i e f i ge
I c ea i g he . f ac hage
W k b a ge i g APC a d ec i ge f
deg ada i (a ge e e i e a ac a ge
be f i e ce i jec i ie b e gh f
e e)
Lead a e effec i e i e e e
I c ea e he i ge ici f eak a ige
E ha ce eed a d d a i fi e e e
CATEGORIES OF ANTIGENS Si aea d d aeh a e e , i c di g
A oan igen : Be g he H a ib d i e
AKA e e e ed a ige Si a e ce - edia ed i i
N i ai fi e e e I ei d ci f c a i i
A i e di ea e - he a a ige E ha ce i e e e i i gica i a e
i aei e e e a ie , a ic a i fa
i.e. P ei e (e e) T dec ea e he d e f a a ige e i ed
Alloan igen : F he e be f he h ecie Red ce c a dei i ae i c e ie
Sa e ecie , b diffe e e /bei g e ie e f b e h
Si ae i e e e M e i i a ici h a
I.e. b d a ige , Ag i ed i i e E ample of Adj an :
a a Al m / Al min m al : C e a ed
He e oan igen : F he ecie adj a i he USA
Diffe e ecie ( a Ag h a ) C e e ihi ge i c ea e
He e ophile an igen : He e a ige ha e i i i e a d e e a id e ca e f
e a ed a a i a b a e ei he ide ica i e
c e - e a ed i c e ha a ib d e ca O he pe : F e nd Adj an
c - eac / he a ige f he he Made f i e a i , e ifie & ki ed
c bac e i
I de f he T-ce ec g i e a a ige , i h d be
ce ed a d b e d i e ide a d e e ce f MHC
*RECAP OF T-CELLS
3 MAIN SUBTYPES
MAJOR HISTOCOMPATIBILITY COMPLEX
T-Helpe cell DESCRIPTION & FUNCTION
ec g i ed b he e e ce f CD4 a ke Majo hi ocompa ibili comple (MHC) - e f
I ed i : (1) ac i a i g ac hage , T a d B ge e ha c de f ce - face ec e ( e i a ige
ce & (2) i f a ai ec g i i )
T-C o o ic cell B i g a ige i he b d he face f ce f
ec g i ed b he e e ce f CD8 a ke ec g i i b T-ce
I ed i : (1) di ec ki i g f he a ge ce Ca f c i a a ige he a a ed
T-Reg la o cell De e i ed he he a a ed i ei
ec g i ed b he e e ce f CD4 a d CD25 hi c a ib e
a ke M hic e f di h a
I ed i : e i g he i e e e BACKGROUND
agai e f a ige e e a eac i f he Di c e ed a a ge e ic c ha de e i e acce a ce
b d . ejec i f i e g af e cha ged be ee e
MHC ge e a e hic a d i a e e a e
PRINCIPAL FUNCTIONS OF T-CELLS c d i a e e ed, i.e:
Defe e f i ace a ic be Fa he : HLA A9, A10 / HLA B4 B8 / HLA CW CZ
I hibi i fi e e e M he : HLA A7 A13 / HLA B5 B9 / HLA CW CX
Ac i a i fi e ce (Mac hage a d B Ce ) Chi d: HLA A7 A13 / HLA B5 B8 / HLA CW CZ
S ec ed fa he i he bi gica
REMEMBER ABOUT T-CELLS fa he
★ De e i he Th U ed de e i e a e a i c i
★ F d i he L h N de , Th acic D c F id Me b a e ei e e APC ha di a ce ed
★ 60-80% f he ci c a i g h c e i he b d a ige be ec g i ed b T-ce
★ I ed i he ada i e i i :E d d c f De i e f he e ea ch a a ai ha a ed i
ac i a i f c ki e he id-20 h ce
★ An igen : CD2, CD3, CD4, CD8 The e a e die hich ided i igh he
★ Pe ide a e b d a d di a ed he MHC ec e f e g e i g he acce a ce ejec i f
APC i e ( Hi c a ibi i )
Re ea che ea fi di g h ha a id ejec i f
a a a de e i ed b a i g e ge e: MHC gene
La e die i dica ed hi ge e a i fac a Comple
A e f c e i ked ge e i he i ed a a i
Nece a f c f ce a i e ac i fi e ce
P d ci f ce ai e ei
HUMAN LEUKOCYTE ANTIGENS
Ce face a ke ha a i e ce di i g i h
ef f - ef
The e a ige e e fi de c ibed WBC ( e k c e )
a d a e c ded f b ge e i he MHC ca ed
Ch e6
O he h a f he ch e6 NOMENCLATURE
MHC ge e d c e e ide ified a e ib e f g af Defi ed e gica h gh e f a ba e f a ib die
ejec i HLA a ige a e a ed acc di g he d c
HLA: Te a c i ed b Da e beca e he e e e fi e e ed b he Ge e L c (Ca i a e e ) a d he A e e
defi ed b di c e i g a a ib d e e ci c a i g (N be )
WBC EX. HLA-A2
A-L c
Ch e 6: 2-A ee
INHERITANCE OF HLA
Haplo pe: C bi a i f i he i ed HLA A e e
T ha e ( ef each a e ) a e a ge e
Beca e f he a ge be f a e e i he MHC, a
e HLA ei a a i e a a fi ge i
U i e e i a be i a a ai
Ge e c di g f HLA i f d he h a
Sh a ha 4 aj a ea a d di ided i 3 diffe e ci
1. Cla I
F d A, B, & C
Nea e e ic a ea
I ed i a ige ec g i i
E e ed he face f a c ea ed
ce
O e ge e c de f each a ic a
ec e
2. Cla II
F d i D egi 2 KEY FUNCTIONS OF MHC IN ANTIGEN PRESENTATION
Maj ci: DP, DQ & DR egi
I ed i a ige ec g i i 1) T elec i el bi d e ide d ced he ei a e
E e ed he face f he APC ce ed i ide he ce f he h
O e ge e c de f a ha chai & e 2) T p e en e ide he face f he h ce a
e ge e c de f be a chai T-ce i h a c ec T-ce ece
3. Cla III
Be ee C a I & II ch e MHC II ec g i ed b T-c ic ce (CD8 a ke )
C de f C4B, C4A, a d C2 MHC I T-he e ce (CD4 a ke )
Sec e ed ei ha ha e a i e
f ci T-ce d e d h ce i he ab e ce f f eig
A c de f D c e e ei e ide
a d ge e c di g f i e ec i T-ce f c e di g i fec ed ce b ca
fac h MHC i a i a i e d h ce ha a e i fec ed
d ci f c ki e a d he ★ The ef e, MHC ha a i a e d i g he
b a ce diffe e ia i f T-ce i he Th (i a e T-ce )
NOT i ed i Ag ec g i i a d i he e e f a e T-ce
NOT e e ed ce face
N e e ed he ce b i a
i i e f c i ( ec e e ce ai
b a ce )
Ha e c e e diffe e c e
c a ed he ca e
Cla III Se ei ec e
C' c e , c e e e e
c ch e 450, C ki e
h d a e a d TNF P ei
MHC C a I MHC C a II
F di c ea ed E e ed APC
ce Wi h he a cia ed
I e ac i h T-c ic e ide i e e i
ce (ce / CD8+ CD4+ T-ce (T He e
a ke ) ce )
ERp57 - a bi d a -chai ha i i
ai ed i h 2- ic g b i
O ce a -chai i b d i h 2- ic g b i , Ca e i &
ER 57 i be e ea ed
Chape one : Cal e ic lin a d Tapa in
The e i j i he c e a d he
abi i i g he MHC ec e f e ide
bi di g
I ace a ei a e dige ed i he p o ea ome
P a - acke fe e f ed i a
c i de h gh hich e ide a a d a e
c ea ed
Pe ide be f ded bef e
e e i g c i d ica cha be
C ea ed e i e f de i e
ca I ec e
Cla I molec le Made f a c e e e ha deg ade ei
Mai e e e ide he i ed i hi he ce e e i he c a
T-c ic (CD8) ce Re ide i a ge c a ic c e e
Wa chd g f i a, a d a a i ic a ige The e ei ca be c ic ei f he
ha a e he i ed i hi he ce (agai f eig b a ce i ca a deg ade
ac a a h ge ) da aged/i e f ded ei a defec i e
Cla II molec le ibo omal p od c (DRIP )
P e e e ge a ige T-he e (CD4) ce P d c f ea e = e ide f ag e
E ge ei ake i he ce f N e: he d bec e i g e a i
ide & deg aded acid ec e
He a i e e e bac e ia Ta ed RER
i fec i he a h ge ide ce Pe ide a e a ed / ed f he c a
( ac a a h ge ) he e f he ER
Ho do he MHC molec le and he pep ide in e ac ? TAP1 & TAP2 (T a e a cia ed i h
MHC c a 1 df he e d ge ah a a ige ce i g) - ecia i ed a e
A ige c i g f i e , a d ei ha de i e e ide f ea e
a a i e a e a ead f d i ide he ce ( a ia ER
dige ed a d a ead f ed i he c a ) M i ab e f ATP-de e de
a f e ide he MHC C a I
CLASS I MHC-PEPTIDE INTERACTION ec e
M efficie a a i g e ide
ENDOGENOUS PATHWAY c i i g f 8-16 a i acid
Ca I ei he i ed i a e ce a c a I RER Whe e he bi d i h e ide
ec e ( h e d ge ) Ta a i b i g i i i he TAP a e
S he i ed i he gh e d a ic e ic a d he MHC C a I ec e
Whi e ai i g f he e ide ( ce ed a ige ), I edia e he i e ac i a d adi g f
c a 1 i e ai a ch ed i he RER he e e ide i he c ef C a I ec e
he i bi d i h he e ide Ra id a ed ce face ce a ha chai bi d
Calne in i h e ide
The 88-kd e b a e-b d ec e i he ER ➢ Cal e ic lin - a he i a cha e e
kee i g he chai a ia f ded hi e i i He i abi i i g he e f ed c a 1
b d i ec e a d he a ia e f di g f he
Thi i be he ca e hi e i i ai i g bi d i h ec e
he 2- ic g b i
"Cha e e" ei ha he i abi i i g he
MHC ec e
POST-TEST
1. Which f he f i g b a ce i he i ge ic?
a. Li id
b. Tef
c. P ei
d. S ga
2. Se f-a ige a e ca ed?
3. (A a g ) A ige : _______ a A ib die : Rece
4. F a a ige be i ge ic i h d ha e a ec a
eigh f a ea ________ da
5. (T/F) A ha e i i e f i a a ige
6. E e ed b a c ea ed ce
7. I ge e f c di g a e f d i ch e6
8. I aei i
9. P ce ed a ige i e c ed i a e ic e
10. MHC- e ide i ec g i ed b a CD8 + T-ce
ANTIBODY
Mai h m al elemen f he ada i e e e
H a - b a ce d ced An igen-binding i e i e ed a a a a e
Te ed a a Imm n gl b lin C ed f fi e aj c a e (ba ed hea chai )
Imm gl b li - g c ei f di e IgG - ga a hea chai
i f he f b d IgM - hea chai
E e ia e i a ige ec g i i & i e IgA - a ha hea chai
e e( i ai &c e e ac i a i ) IgD - de a hea chai
C i e 20% f a a ei i hea h IgE - e i hea chai
i di id a
N e: A a ib die a e c ide ed Ig ; b a
i g b i a e a ib die
MONOMER
Ba ic c a i fa i g b i
DISULFIDE CHAINS: S abi i e 3D c e fi g b i
Each e ha 2 a ai ab e A ige ic Bi di g Si e = 2
(N e: -c ale b d al hel i abili i g he c e)
Fab egi
In achain S abi i e he d ai (g b a /ba ike
Valence N . f c bi i g i e
egi ) f he e ide chai
M e - 2 bi di g i e
In e chain B d f d be ee (1) Hea a d igh
Pe a e - 5 e 2 bi di g i e = 10 a
chai a d (2) T hea chai
An ib d Valenc N . f ece i e f a ige
E i e f d i he a ige hich i bi d he
ac i e i e f he a ib d
An igen Valenc N . f a ige ic de e i a hich
e e a bi di g i e f a ib die
2 MAIN REGIONS
Va iable egi n U e a f he a ib d
U ed f bi di g f he a ige
C i e a i - e i a e d f chai
C n an egi n L e a f he a ib d
U ed f bi gica ac i i ie f IG ( i ai ,
bi di g f b h ece a dc e e f ag e )
C i e f ca b - e i a e d f chai
Di ided i 3 egi hea chai
Gi e a a e f each i g b i e
N e: Hinge egi n A he f e ibi i f he N e: I he c a egi he e he ca b g i f d, he
i g b i beca e i i ich i i e ca b h d a e i i ca ed (be ee CH2 a d CH3 a ea)
Abi i be d e 2 a ige -bi di g i e eae Ca b h d a e f nc i n :
i de e de & e gage i a g a i 1) I c ea e bi i f he a ib d
F e ibi i a i i effec f c i (i c de 2) P ec i f deg ada i
i i ia i fc e e ca cade & bi di g ce ih 3) E ha ce he f c i a ac i i f he Fc d ai f
ecific ece f Fc i ) he a ib d
F d i be ee CH1 (c a egi f he 1 I a beca e g c a i a ea
d ai ) a d CH2 (c a egi f he 2 d d ai ) be c i ica f ec g i i b Fc ece
a ea (f d hag c ic ce )
Si e he e he e e ac
Regi n De c i i n Im ance
P e e i gamma, al ha, & del a chain
Ab e i &e i chai VH a iab e egi f H chai Bi di g i e f a ige he e
h e a iab e egi ae
Va iable egi n C n an egi n
f d (CDR1,2,3)
U e a f a ib d L e a f a ib d
VL a iab e egi f L chai Bi di g i e f a ige he e
C i f 110-120 a i acid f Ha i i a a i acid e e ce h e a iab e egi ae
b h H a d L chai ha a ie f ( i i 111 ad )a g each f d (CDR1,2,3)
ei g b i ca a he e Ig
CH1 c a egi he Bi di g i e f C4b
Se e ce f A.A. c i i g f Se e ce f A.A. c i i g f 1 d ai f H chai (c e e f ag e )
e ide chai change e ide chai i fi ed and
CH2 c a egi he Bi di g i e f C1
nchanging
2 d d ai f H chai ( ec g i i ec e)
Amin g (NH2) i e i a Ca b lg (COOH) i
CH3 c a egi he Bi di g i e f he Fc
e d e i a e d
3 d d ai f H chai ece f di c e
F ci : binding i e f Ag F c i : e a ed he bi l gic a d ac hage .
ac i i ie f Ig ec e A he e hag c e (B&T),
a a ce , he e g
Incl i n : VH a d VL, c ec i e Incl i n : CL, CH1, CH2, a d a ce a ach
e ed a Fab egi n (a ea he e CH3
a ige bi d ) CL c a egi f L chai -
H e a iable Regi n : Di c e e /f e i e
jec i g a d f he e i a e d f he a iab e L chai .
Thi i he i e he e he e i e bi d
CDR (c m leme a i de e mi i g egi )
Occ a i f d f a iab e egi
f b h chai
C i f CDR1, CDR2, CDR3
A ige bi d i idd e f CDR i h a ea f f
CDR i ed
Ag i ca ed i hi he f d b bi di g a he e
h e a iab e egi
ANTIBODY FRAGMENTS
Red c i b Me ca e h a i e
Me ca e ha ha d f he
b eakd he a ib d b de i g
di fide b d
Re l : 2(Fab)2 + Fc
Fab 1 L chai a d H chai
Remembe hei c m i i n:
Fab 2 Ligh chai a d 2 ha e f Hea chai
Fc 2 ha e f he Hea chai
ALLELIC EXCLUSION
DEVELOPMENT OF ANTIBODIES
P ce he eb he ce i c i ed he e e i fa
Acc di g : SUSUMU TONEGAWA a ic a V egi f i hea chai a d i igh chai
Ch e c ai i ac Ig ge e . Di e i i he Ig Occ f i g a cce f ea a ge e f he Ig DNA
ec e i ge e a ed a a e f a ic ec bi a i f eg e
ge e
I e ge e ea a ge e d ce ecific a ib die
FUNCTIONS
P ide i i e b a ib d ha ca c
ace a
Fi i g c e e ac i a i
C a i g a ige f e ha ced hag c i ( i ai )/
Faci i a e i ai
Ne a i a i f i a d i e
Pa ici a i i agg i a i a d eci i a i eac i
Be e a eci i a i eac i (i e a
b e a ic e ch a IgG)
ADCC
IMMUNOGLOBULIN G (IgG)
IMMUNOGLOBULIN M (IgM)
M ed i a /ab da i g b i ca
75-80% f e Ig Bigge i g b i (AKA Mac gl b lin )
*70-75% f a e Ig [acc di g S e e ] Half life: 6-10 da
Maj Ab i ec da i e e e M lec la c e: Pe a e
MW = 150,000 (7S) Highe i e i i a i e e e
L nge half-life (a 23-25 da ) 5-10% f e i g b i
S a e i g b i ec e MW = 970,000 (19S)
Ha 4 bcla e ( ai diffe i be & ii f 1 a ea af e A ige ic S i ai
di fide b idge be ee ga a chai ) C ai 10 f c i a bi di g i e
IgG1 (66%) - e d ei a ige I c de e ec a d ai f d ga a chai
G d a i i ia i g hag c i bi d F :
g Fc ece Pe a e ic = ec e ed i e
IgG2 (23%) - e d accha ide a ige M e ic = B ce face
Ca c ace a J (j i i g) chai
IgG3 (7%) - ei a ige H d ge he fi e e ic i
La ge hi ge egi ( be f G c ei ade i a a ce ha c ai
i e chai di fide b d ) efficie e e a c ei e e id e
a bi di g c e e Se e a i kage i f di fide b d be ee
G d a i i ia i g hag c i bi d adjace e
g Fc ece
Ma i i ia e e i a i & faci i a e ec e i a
c a face
O e J chai e e a e
M e efficie ha IgG i c e e ac i a i
F d i i a a c a a d i he b d f id
i e
Ca c he ace a
U ed diag e ac e i fec i
N e ce e i f IgM
M e efficie i agg i a i eac i
FUNCTIONS
C e e fi a i IMMUNOGLOBULIN A (IgA)
Agg i a i
O i ai Re e e 10-15% f a ci c a i g i g b i
T i e ai ai MW = 160,000 (7S)
S face ece f i a e B ce P ed i a Ig i ec e i
M efficie i ac i a i g he c a ica c e e ah a B d Sec e i - I a Di e
(i i ia e eac i f i e bi di g i e ) An i e ic Pain
Na a I he agg i i - ABO a ib die S he i ed a highe a e ha IgG
Ha 2 bc a e
1) Se m IgA / IgA 1
Mai e ic
P e ib e h gh c
2) Sec e IgA / IgA 2
P ed i a f i ec e i a c a face
S he i ed i a a ce f d ai i MALT
Ha 13 a i acid e ha IgA1
M e e i a e bac e ia
ei a e ha c ea e IgA1
Di e i h J Chai a d ec e c e
I e Re e A ea i b ea i k, c , a i a, ea &
ea
★ NOTE: IgA ca ab e f C a ica C ac i a i
An i-inflamma agen
D - eg a e IgG- edia ed
hag c i , che a i , bac e icida
ac i i a d c ki e e ea e
Pa c a face
Ne a i e T i
FUNCTIONS OF He e e bac e ia adhe e ce
SERUM IgA c a
Ca ab e f ac i g a i (bi di g
IgA- ecific ece e hi ,
c e & ac hage igge
e ia b & deg a a i )
Pa c a face a d ac a a fi
i e f defe e
Ne a i a i f i
P e e bac e ia a d i a adhe e ce
c a face
IMMUNOGLOBULIN D (IgD)
P ide a a a i e i i
e b h gh b ea feedi g Smalle c ncen a i n i e (a 0.001%)
Ca ab e f ac i g a i M aef d face f ai e B L h c e
Be ie e a a e i eg a i g B-ce a ai a d
diffe e ia i
Half life: 1-3 da
MW = 180,000
E e ed i a ed B ce face
Sec d e fi g b i a ea
FUNCTIONS OF Idea ea e de a ige
SECRETORY P a a e i B cell ac i a i n, ma a i n and
IgA diffe en ia i n
Ha a g hi ge egi e ce ib e
e i
D e bi d c e e , e hi ac hage
IMMUNOGLOBULIN E (IgE)
Lea ab ndan in e m (a . 0.0005%)
MW = 190,000 (8S)
N ca ab e f C fi a i , agg i a i , i ai ,
c i g he ace a
N i ance Ab
Reagi ic a ib d
Pa ici a e i i edia e h e e i i i ie eac i (EX.
A h a, A a h a i , Hi e )
P d ced b a a ce a g a d i ki
A ache he f i g af e he i h gh Fc R (CH3)
Ba hi
La ge ha ce
E i hi
Ti e a ce
M hea - abi e f a i g b i
Hea i g a 56 C f 30 i -3 h
c f a i a cha ge & e abi i bi d
a ge ce
Bi di g ca e deg a a i (hi a i e & he a i e ea e)
Ca e eIi edia e h e e i i i (a e gic
eac i ) ha fe e , a h a, i i g, dia hea,
hi e & a a h ac ic h ck
FUNCTIONS
Abi i ac i a e a ce & ba hi
P a a aj e i a e gic eac i
Vi a i e i i a i i a a i e , e ecia he i h
T igge ac e i f a a eac i ha ec i e hi
&e i hi ie fi fa ai
TYPES OF ANTIBODY DIVERSITY
Defi ed b he c a egi f he H
chai
The H chai ha i i e each Ig c a
U i e a i acid e e ce c
a Ig ec e f gi e c a i gi e
ecie
Ha e diffe e hea chai
Re e e c a e f a ib d
ISOTYPES
PROPERTIES OF IMMUNOGLOBULIN
Ha e he a e c a egi ih
i ,b i gic diffe e ce
Ge e ic a ia i i he c a egi
Ba ed he diffe e ce i a i acid a a
a ic a ii he c a egi f
he H L chai
Occ i 4 IgG bc a e , 1 IgA bc a ,
a d i ka a igh chai
Diffe e i di id a ha e diffe e a e
T in c i e he ie la ed a e a f ll : Thi i ac i a e he ece a d
1. DIRECT TEMPLATE THEORY he e i a e d ci f he
Fi a ed b B einl and Ha i (1930) a e e f ece ha
Pa ic a a ige a ige ic de e i a d ci c a e a a ib die
e ea a e a e agai hich a ib die d (C e a Mecha i )
f d T ke e i e :
The a ib d ec e d he eb a ea L ck-a d-ke c ce f
c fig a i c e e a a ige e ae he fi f a ib d f
2. INDIRECT TEMPLATE THEORY a ige
Fi a ed b B ne and Fenne (1949) A ige e ec ed ce ih
The gge ed ha he e f a ige ic b i -i ca aci e d
de e i a i he a ib d - d ci g ce i
i d ced a he i ab e cha ge i he e ce . A
Je ne (1955)
ge c f he a ige ic de e i a a
D i g he e b ic ife, i i
i c a ed i ge ea d a i ed he
fg b i ec e e e f ed
ge ce /
agai a ib e a ge f
SELECTIVE THEORY OF ANTIBODY PRODUCTION a ige
NATURAL The a ige , he i d ced
Acc di g hi he he i c e e ce ha e a
SELECTION THEORY he h , c bi e e ec i e ih
e ic ed i gica a ge
(1955) he g b i ec e ha ha he
A ige i a e he i c e e ce e ec i e
ea e c e e a fi
he i e a a ib d
The g b i i h he c bi ed
i.e. a he ge e ic i f ai i e e i he ce
a ige i a ed a ib d -f i g
bef e e c e i g he a ige
ce d ce he a e e f
The ce ee d ci g a ib d a e e b a ige ic
a ib d
i ai e i d ci f e ec i e a ib die
B ne (1957)
P i a ige e e, ce ai ce
I di id a h c e ae
a ead e ecific face
ge e ica e ga ed
ece
d ce e e f
O ce a ige i i d ced, i i
i g b i
e ec i a ia e ece
A ecific a ige fi d e ec
The bi di g be ee he a ige
h e a ic a ce ca ab e f
a d he ece ead b eaki g
e di g i , ca i g he
ff he ece a a ib die
ife a e
hich he e e he ci c a i
Ne ece df i ace
f b ke ff ece
CLONAL SELECTION
EHRLICH S HYPOTHESIS
SIDE-CHAIN (1957)
THEORY
(1898)
A a ge be f c e f
i gica c e e ce
I -c e e ce ha e bea i g ecific a ib d a e
face ece ha a e ca ab e ae d ced d i g fe a
f eac i g i h a ige , hich de e e b a ce f
ha e c e e a ide chai a ic ai fi gica
S ecific a ige fi he e
c e e a
c e e ce (ICC) agai a T e a a e ge e c de f he
ib e a ige c a a d a iab e egi f
A i di id a ICC e e e i g b i
RECOMBINATIONAL
e b a e ece ha a e The e c d be a a be
GERMLINE THEORY
ecific f a di i c a ige c di g f he c a egi a d
Thi i e ece a a ge be c di g f he
ecifici i de e i ed a iab e egi
bef e he h c ei
The b e ed e e i e i
e ed a ige
ge e a ed f a i i ed be f
Bi di g f a ige i ecific SOMATIC
i he i ed V- egi e e ce
ece ac i a e he ce a d VARIATION THEORY
The de g a e a i i hi B
ead ce a ife a i f
ce d i g he i di id a ife i e
c e , he i i g he a ib d
M idel acce ed he
P ide f a e kf MONOCLONAL ANTIBODIES
be e de a di g f
he ecifici , A ib die de i ed f a i g e a e a ib d - d ci g
i gica e , ce ha ha e d ced a i e , h f i gac e
a d he e f De e ed f diag ic e i g f B ce ha he i e
ec g i i f efa d e ecific a ib die
- e f b ada i e PRODUCTION OF MONOCLONAL ANTIBODIES
i i
D a back: c ide a i f ge e ic
ba i f di e i f a ib d
ec e
D e e & Be e (1965)
ed ha c a & a iab e
i c ded f b e a a e
ge e
APPLICATIONS
Diagn ic Te
Ab a e ca ab e de ec i a ( g/ L) f
ec e
EX. P eg a c H e
Diagn ic Imaging
Ab ha ec g i e a ige a e adi abe ed
i h i di e I-131
Imm n in
Ab c j ga ed i h i
mAb clea a h gen
HYBRIDOMA TECHNIQUE
De e ed b Ge ge K h e a d Ce a Mi ei ( 1975) -
di c e ed ech i e d ce a ib die a i i g f
i g e B ce
*H b id ma - f i f diffe e e f ce
U e f i f a ac i a ed B ce i h a e a ce ha ca
be g i defi i e i he ab a
HYBRIDOMA PRODUCTION
A ei i i ed i h a a ige
The ee i ha e ed a d c bi ed i h e a ce
ih e h e e g c (PEG) a fac a
PEG - b i g ab f i f a a ce ih
e a ce
Ce a e he aced i c e edi ihh a hi e,
a i e i , a d h idi e (HAT edi ), hich i e ec i e
f h b id a ce
HAT edi - e aae h b id a ce
(a i g g e ec i e & a i g f ed
e a ce f ed ee ce i e)
M e a ce ca he i e
c e ide die
N a B ce ca be ai ai ed PRIMARY and SECONDARY IMMUNE RESPONSE
c i i ce c e die
H b id a ce a e di ed a d aced i ic i e e PRIMARY IMMUNE RESPONSE
g L g ag ha e
Each e (c ai e c e) i c ee ed f e e ce f S e e ia i c ea e i a ib d
de i ed a ib d b e i g e aa Sh i ed e e
T-CELL RECEPTOR
O f d he T ce e ba e
C ed f 2 e ide chai , (a ha) a d (be a)
G c ei ec e i ade fac a a d a iab e
egi hich c i e he a ige -bi di g ie
S e TCR e e a (ga a) a d a (de a) chai
The e ha e ecifici f c e i a a ige ,
ch a hea a d h ck ei h h i id
T-CELL DIFFERENTIATION
Th m c e
L h c e i he Th
Ma ke : CD44 a d CD25
STAGES
1) D ble-Nega i e S age
Th c e ha ack CD4 a d CD8
P ife a e i he e c e de he
i f e ce f IL-7
2) D ble-P i i e S age
Whe he h c e e e b h CD4
a d CD8
T-HELPER SUBSETS
Th1 - Agai i ace a a h ge
Th2 - Aid B ce d ce a ib die
Th9 - Ha e i fa a effec
Th17 - I c ea e i f a a i a d j i de ci
T eg - S i ch ff i e e e
MATURE T-CELLS
S i f e ec i ce e
E hibi e e f a ke :
1) CD4 + T-cell
AKA T Hel e T Ind ce cell
I e ac i h a ige a d MHC II ei
2) CD8 +T-cell
AKA T C ic cell
I e ac i h a ige a d MHC I ei
ACTIVATION
POSITIVE OF T-HELPER
SELECTION OF CELLS
THYMOCYTES
B-CELL DIFFERENTIATION
P -B Cell
Ma ke : CD19, CD45R, CD43, CD24, c-Ki
P e-B Cell
NEGATIVE
Wi h chai i c a a d e i e i ce
SELECTION OF
face
THYMOCYTES
Ma ke : CD19, CD45R, a d CD24
Imma e B Cell
Di i g i hed b he a ea a ce f c e e IgM
ec e he ce face
Ma ke : CD19, CD45, CD24, CD21, CD40, a d
MHC II ec e
Ma e B Cell
Ha e IgM a d IgD i face f a ach e f
a ige
Ei he a gi a e B ce F ic a B ce
Pla ma Cell
Re e e f diffe e ia ed h c e
Mai f c i i a ib d d ci
Pla ma Cell
Ma e B Cell
BCR TCR
ROSETTE
TECHNIQUE
POST-TEST
1) Which f he f ll ing e e a a ignaling ein
nce an an igen i a ached he TCR?
USE OF FICOL a) CD2
HYPAQUE b) CD3
c) CD4
d) CD8
2) (T/F) D ble i i e h m c e in he h mic c e
nde g a i nce he eac ngl
elf-an igen f MHC
3) Which f he f ll ing cl e f diffe en ia i n i ed
iden if T eg la cell ?
a) CD4 a d CD25
b) CD8 a d CD16
CELL FLOW
c) CD4 a d CD16
CYTOMETRY
d) CD8 a d CD25
4) T c ic cell all a ge hich f he f ll ing?
a) Vi a i fec ed ce
b) T ce
c) Bac e ia a ige
d) A a d B
5) (T/F) An igen m be ce ed b APC i
ec gni i n f B cell ece
6) A ce he eb an an ib d facili a e
hag c i f ea ie eng lfmen f he an igen
7) The m efficien an ib d cla ac i a e he
cla ical a h a f c m lemen em
8) The m ed minan an ib d cla ec nda
imm ne e n e
9) The nen f he cl nal elec i n he f
an ib d di e i
10) Which f he f ll ing c m nen i n incl ded in
he medi m f ha e ing h b id ma cell ?
a) H a hi e
b) A i ei
c) Th idi e
d) P e h e e g c
Autocrine action
Relea e of c okine o ld affec he gene a ing cell
( elf); p omo e p olife a ion o he ac ion of ha
pa ic la cell
E .: IL-1 inc ea e ac i a ion of
an igen-p e en ing cell
Paracrine action
Relea e of c okine o ld affec he nea b cell
Affec cell in he immedia e icini
E .: IL-1 can be elea ed b T helpe cell,
ac i a e ne ophil , decolo i e ne on
Endocrine action
Affec cell ha a e fa o emo e f om he
gene a ing/ ec e ing cell
C okine a e of impo ance in infec io di ea e fo o The c okine o ld pa h o gh he blood
con a ing ea on : e el fo ci c la ion o a d he a ge cell in a
The can con ib e o he con ol of infec ion emo e place
The can con ib e o he de elopmen of E .: IL-1 ec e ed in o he blood e el and
pa holog (i.e. ep ic hock). go o a d he h po halam o p omo e fe e ;
im la e he li e cell o p od ce ac e-pha e
eac an
C tokine Receptor
[Refe ab e a he e d f he e e e ]
FUNCTIONS:
1. Con e e acell la ignal , namel he p e ence of
pecific c okine , in o an in acell la ignal, ch a
ac i a ion of an en me ha can igge a a ge cell
e pon e.
2. The a e an memb ane p o ein , and he e acell la
domain bind c okine, he eb p o iding mean of
de ec ion of he e acell la ignal.
INTERFERONS
The be e abli hed an imic obial c okine .
In e fe e i h i al eplica ion
i i he pe I IFN (IFN- and IFN- ) ha f nc ion
p ima il in hi manne
c c
De i ed in 1957 he e i infec ed cell ec e ed a
Multichain receptor comple es
molec le ha in e fe ed i h i al eplica ion in b ande
cell .
Th ee (3) pe : alpha, be a, gamma
T o pe of en me ind ced an i- i al a e:
P o ein kina e
2 ,5 -oligoaden la e n he a e
IFN i mainl a T cell p od c and p od ced la e , and O e ec e ion can lead o ep ic hock
mo n ed NK cell . La ge amo n of TNF ec e ed in
In e fe on al o inhibi i a embl a a la e age, e pon e o g am nega i e bac e ial
con ib e o he an i i al a e: infec ion , ca ing a dec ea e in blood
Enhance cell la MHC pe e (h po en ion), ed ced i e
Ac i a e NK cell and mac ophage pe f ion, and di emina ed in a a c la
Inhibi i e i ho damaging he ho cell coag la ion
DIC ncon olled bleeding
dea h
P omo e a odila ion & inc ea ed a c la
pe meabili
TNF- ac i i i a lea pa iall eg la ed b
ol ble fo m of bo h TNF ecep o
The e ecep o ac o bind e ce TNF-
and, combined i h he ho half-life of
he ol ble fo m, e e o limi he
c okine ignaling ac i i TNF- ac i i
i a lea pa iall eg la ed b ol ble
fo m of bo h TNF ecep o
TNF- a ell a IL-1 a e p e en in he ma oid
no ial fl id and no ial memb ane of pa ien
i h he ma oid a h i i (RA)
TNF- i he cen al media o of
pa hological p oce e in RA and o he
inflamma o illne e ch a C ohn
TUMOR NECROSIS FACTOR di ea e, lce a i e coli i , and j enile
The p incipal media o of he ho e pon e o a hii
g am-nega i e bac e ia-lipopol accha ide componen
(LPS) in bac e ial cell all ha pla a ole in he e pon e TUMOR NECROSIS FACTOR RECEPTORS
o o he infec io o gani m . TNFR1
LPS in lo do e ac a a pol clonal ac i a o of B cell , P ima media o of TNF- an d c ion in mo
con ib e o he elimina ion of bac e ia. cell pe
High concen a ion LPS ca e inj , DIC, and hock Bind he ol ble fo m of TNF-
e l ing o dea h Sh art man reaction Con i i el e p e ed on mo i e and bind
Fi een and i ola ed f om he l mphoc e and ol ble TNF- .
mac ophage TNFR2
When he di co e ed hi , TNF ind ce Ac i a ed b he memb ane-bo nd fo m of TNF-
de c ion o l i of mo cell U all e p e ed in epi helial cell and cell of he
TNF- mo p ominen membe imm ne em
Con i of a lea 19 diffe en pep ide
E i in bo h memb ane-bo nd and ol ble fo m
and ca e a odila ion and inc ea ed
a ope meabili
T igge ed b LPS (lipopol accha ide G (-)
bac e ia)
Effec : a odila ion, a c la
pe meabili , and T cell ac i a ion
Sec e ed b ac i a ed monoc e and
mac ophage (fo nd in he memb ane )
Al o fo nd in ol ble fo m in ec e ion
Ac i a e T cell ind ce MHC Cla II
e p e ion, a c la adhe ion molec le , and
chemokine
Onl one IL-6 ecep o ha been iden ified ha The e molec le ha e he abili o im la e he le koc e
con i of IL-6R ( he IL-6- pecific ecep o ) and mo emen (chemokine i ) and di ec ed mo emen
gp130 ( he common ignal- an d cing ecep o (chemo a i );
b ni ed b e e al c okine ). Reg la e he chemotactic acti it of le koc e
Binding of IL-6 o he IL-6R ind ce ( e pon e o infec io di ea e , a oimm ne
dime i a ion of gp130 i h he - b ni inflamma ion, cance , and homing l mphoc e o
Homodime i a ion confo ma ional l mphoid i e)
change in gp130 ha e po e o ine E .
e id e CCL2 o MCP-1 (monoc e chemoa ac an
Se ie of pho pho la ion eac ion ind ced ignal p o ein 1)
an d c ion b one of he Jan kina e (JAK): A ac ing monon clea cell o i e of
CRP ch onic inflamma ion
Complemen fi a ion (C3) CCL3 o mac ophage inflamma o p o ein 1-alpha
Fib inogen ac i a ion A ac ing mac ophage , monoc e and
IRF-1 ne ophil
T and B cell a e ned on CCL4 o mac ophage inflamma o p o ein 1-be a
U ed o a ac NK cell , monoc e and
o he imm ne cell
IL-8 o CXCL8
A ac ing ne ophil in ac e inflamma ion
Ac i a ing monoc e
CX3
Cell adhe ion ecep o capable of a e ing
cell
TGF-
F nc ion a bo h an ac i a o and an inhibi o of
p olife a ion, depending on he de elopmen al age of he
affec ed cell
Reg la e he e p e ion of CD8 in CD4-CD8- h moc e
Ac a an a oc ine inhibi o fac o fo imma e
h moc e
Inhibi he ac i a ion of mac ophage and g o h of man
diff oma ic cell pe
F nc ion a an an i-inflamma o fac o fo ma e T cell
T 1 cell a e CD4+ T cell ha a e ind ced f om Ke c okine ha diffe en ia e T cell o main ain hem a
an igen-ac i a ed na e T cell in he p e ence of IL-10. Th17 cell a e TGF- and IL-6
E e pp e i e ac i i ie on bo h Th1 and Th2 In e le kin-23 p od ced b mac ophage and dend i ic
b p od cing mo e IL-10, TGF- , o IL-35 cell
IL-10 Pla a ole in finali ing he commi men o Th17
Ha p ima il inhibi o effec on he imm ne cell
em IL-17 po en p oinflamma o c okine and ind ce
P od ced b monoc e , mac ophage , CD8+ T e p e ion of TNF- , IL-1 be a and IL-6
cell , and Th2 CD4+ T cell p od ce p oinflamma o media o f om m eloid
Ha an i-inflamma o and pp e i e effec on and no ial fib obla and pe pe a e he
Th1 cell inflamma o p oce in RA
Inhibi an igen p e en a ion b mac ophage and IL-17A fi IL-17 iden ified
dend i ic cell O he IL-17 famil membe incl de IL-17B, IL-17C, IL-17D,
T-cell pp e ion occ h o gh IL-10 IL-17E, and IL-17F.
inhibi ion of p oinflamma o c okine Mo of he IL-17 c okine famil membe a e po en
and inhibi ion of co im la o molec le p oinflamma o c okine and ind ce e p e ion of TNF- ,
e p e ion on APC . IL-1 , and IL-6 in epi helial, endo helial, ke a inoc e,
Inhibi IFN-gamma p od c ion ia he pp e ion fib obla , and mac ophage cell .
of IL-12 n he i b acce o cell and he IL-17A and IL-17F ind ce epi helial cell , endo helial cell ,
p omo ion of a Th2 c okine pa e n and fib obla o p od ce CXC ligand 8 (CXCL-8)
An agoni o IFN-gamma C cial fo ec i men of ne ophil o he i e of
Do n eg la o of imm ne e pon e inflamma ion.
TGF- Ac oge he i h g an loc e mac ophage CSF o
Do n eg la e he f nc ion of APC and block p od ce CXCL-8 in mac ophage ignal
p olife a ion and c okine p od c ion b CD4+ T ne ophil o he i e
cell D eg la ion of Th17 cell b e and ec e ed c okine
IL-35 pa hogene i of e e al inflamma o and a oimm ne
Imm no pp e i e effec on Th1,Th2, and Th17 condi ion
P omo e T eg A hma and alle gie : inc ea ed Th17 cell and IL-17A
Do n eg la e imm ne e pon e and p e en IL-17A di ec l ind ce IgE p od c ion b B cell
ch onic inflamma ion Highe amo n of IL-17A and IL-17F mo e
e e e a hma
O he pe :
Ind ced T eg (iT eg )
HEMATOPOIETIC GROWTH FACTORS
Can de elop f om ma e T cell in he pe iphe
IN INNATE AND ADAPTIVE IR
TR1 cell
Effect of colon stimulating factors on gro th and
differentiation of blood cells:
Th17 CYTOKINES IN INNATE AND ADAPTIVE IR
G o h of hema opoie ic em cell (HSC) eq i e em cell
Bo h inna e and acq i ed Imm ne e pon e fac o (SCF) i h h ombopoie in (TPO).
Th17 ec e e IL-17 famil of c okine SCF (c-ki ligand) In e ac i h c-ki
C cial in bac e ial and f ngal infec ion , e peciall Needed o make BM em cell e pon i e o o he
in he m co al a ea CSF
Upon enco n e i h bac e ia o f ng SCF in i elf canno im la e BM
APC ec e e c okine diffe en ia e p ogeni o cell
Th17 b e of cell . Impo an ole in main aining iabili and
Th17 cell in ade he infec ed a ea and p olife a i e capaci of imm ne cell ch a
ec e e IL-17 c okine nece a fo imma e T cell and ma cell
con in o ec i men of ne ophil Colon Stimulating Factors im la e fo ma ion of
Th17 in local i e, ma ha e an impo ance in long- e m colonie of cell in he bone ma o
main enance of an imic obial e pon e o IL-17 c okine P ima media o of hema opoie i
im la ion, ec e e an imic obial pep ide . IL-3: ind ce BM em cell o fo m T and B cell
GM-CSF: diffe en ia ion o o he WBC pe
A hii
Defec in he IL-1RA
Unde e p e ion p omo e f he inflamma ion
Inflamma o Bo el Di ea e
C okine in ol ed a e a ocia ed i h p omo ing
inflamma ion
Dem elina ing nd ome
The m elin co e ing he ne e fibe a e de o ed
o ed ced
CYTOKINES
FACTOR SOURCE ACTIONS
IL-16 CD8 T, CD4 (no p efo med) Chemo a i CD4 T cell and eo inophil
IFN M l iple An i i al
IFN M l iple An i i al
T cell An i i al, ac i a ion of mac ophage & inhibi ion of TH12 cell
IFN
NK cell MHC ind c ion
TGF T cell / Mac ophage Inhibi ac i a ion of NK cell , and T cell , mac ophage ,
inhibi p olife a ion of B and T cell
ANTIGEN-ANTIBODY REACTION
Imm ne eac ion occ ing be een he antigenic
determinant (Epitope) and Fab (Antigen binding
fragment) of he an ibod molec le
An an igen ma con ain onl one ( ni alen ) o
mo e han one epi ope (m l i alen )
Fab region de e minan fo nd in he an ibod
Re pon ible fo Ag ecogni ion
[RECALL]
Fc po ion of Ab biologic
p ope ie of Ab (e.g. binding fo
complemen )
The imple combina ion of a Ag and i
co e ponding Ab i called sensiti ation LAW OF MASS ACTION
Fo ma ion of imm ne o Ag-Ab comple
If he an ibod ha pecifici o he
co e ponding epi ope, he epi ope o ld
hen bind o he co e ponding Fab egion,
fo ming an imm ne comple .
AVIDITY
Specificit The abili of an indi id al an ibod
combining i e o eac i h onl one an igenic de e minan
The abili of a pop la ion of an ibod molec le o
eac i h one an igen onl
A pecific Ab i o ld onl eac o a pa ic la
epi ope p e en in he Ag
The an igen can be compo ed of m l iple epi ope
ae e e
Tha epi ope p e en can be
O e all binding i e -de e a o -de e a
O e all eng h of an igen an ibod binding and i he m Uni-determinant all of ho e epi ope
of he affini ie of all he indi id al an ibod an igen o ld ha e he ame pecifici
combining i e Multi-determinant m l iple epi ope
S eng h i h /c a m l i alen Ab bind a m l i alen Ag i h diffe en pecifici ie ; no all ho e
Mea e of o e all abili of an Ag-Ab comple epi ope o ld bind o a pecific/pa ic la
A high a idi can ac all compen a e fo a lo affini Ab
Mo e bond be Ag and Ab = highe a idi en i i e, pecific
[RECALL]
Each monome of he an ibod o ld ha e 2 binding Fal e po i i e a e p e alen
ie . P obabl ha e c o eac ion
Compa ing he IgG Ab (10 6) . IgM Ab (10 10), he IgM o ld The Ab migh bind o o he Ag hich a e
ha e a highe a idi d e o i m l iple binding i e . i e imila o o iginal Ag
S onge comple Cross Reactivit
Le likel o di ocia e Impo an in affini
A idi ma al o be a ocia ed i h he fo ma ion of la ice, Abili of an indi id al Ab combining i e o eac
fo e ample, in aggl ina ion eac ion . i h mo e han one an igenic de e minan
The IgM Ab o ld be a be e /e cellen aggl inin The e a e ce ain ca e he ein he Ab i
compa ed o IgG beca e i o ld ha e mo e able o bind i h no j one pecific
binding i e o allo he pa ic la e m l i alen epi ope, b co ld al o bind o o he
aggl inogen o bind epi ope ha omeho ha e he ame
IgM ha he po en ial o bind o 10 diffe en Ag cha ac e i ic o pa ha a e i e
IgG onl 2 binding i e , malle in i e imila o ela ed
Al ho gh i i capable of binding o Ag, The mo e he c o - eac ing an igen e emble he
he e o ld be no i ible eac ion o o iginal an igen, he onge he bond ill be
aggl ina ion be een he an igen and he binding i e.
The abili of a pop la ion of Ab molec le o eac
i h mo e han one Ag
INTERMOLECULAR FORCES
An an igen can be:
The econd ep
Univalent/Monovalent 1 epi ope p e en
Fo ma ion of c o -link ha fo m he i ible agg ega e
Multivalent he e a e mo e han one epi ope p e en in
Rep e en he abili a ion of Ag-Ab comple e i h he
he Ag molec le
binding of m l iple an igenic de e minan
In mo clinical ca e / pecimen
A he la ice i fo med, he imm ne comple become
Uni-determinant m l iple epi ope ha e he ame
la ge , and e en all , a i ible e l can be een.
pecifici
La ice h po he i fo m la ed b Ma ack
Multi-determinant m l iple epi ope b do no ha e he
Fi a hi in p ecipi a ion eac ion
ame pecifici o a pa ic la Ab
Ba ed on hi a mp ion, each Ab molec le
ho ld ha e a lea 2 binding i e , and he Ag
LEFT m l i alen an igen,
m be m l i alen in o de fo he la ice o fo m.
m l i-de e minan epi ope
Fo e ample:
RIGHT m l i alen an igen,
Ab i h 2 binding i e + m l i alen Ag
ni-de e minan epi ope
Once he e bind, en i i a ion occ .
C cial o he
B ince he e a e o he binding i e a ailable,
acc ac of e ologic e
he o he binding i e of he Ab i ill a ailable, o
ano he Ag can bind he e.
SENSITIZATION O he binding i e of he Ag a e ill a ailable, o
ano he Ab o ld bind he e oo.
E en all , he la ice ill fo m and o can no
ee a i ible e l .
La ice fo ma ion o he i ible e l o ld al o be affec ed
b he n mbe of Ag and Ab p e en in he
ol ion/ pen ion.
In o de fo he la ice o fo m, he e m be a
p opo ional amo n of Ag and Ab (no e al, b
A men ioned, en i i a ion i he fi pha e, hen he
he p ope e m i p opo ional)
epi ope bind o i co e ponding Fab
Sen i i a ion can happen e en i ho he
Aggl ina ion, like p ecipi a ion, i a o- ep
p opo ional amo n of Ag and Ab, b he eac ion
p oce ha e l in he fo ma ion of a able
ma no be een.
la ice ne o k. The fi eac ion, called
Nagbab d, ag e- e e, e ce
en i i a ion
e he ba g Ag ba g Ab,
IMMUNOASSAYS
Imm noa a a e ick and acc a e e ha can be
ed on- i e and in he labo a o
De ec pecific molec le and/o mea e he
concen a ion of a molec le in a ol ion h o gh he e of
an an ibod o imm noglob lin
Imm noa a el on he inhe en abili of an an ibod o
bind pecific c e of a molec le efe ed o a "anal e"
and hich chemicall in man ca e , a p o ein.
GENERAL CONSIDERATIONS
In an imm noa a , an an ibod molec le
ecogni e and bind o an an igen
Binding i ela ed o:
Concentration of each reactant ( onal
eac ion )
Specificit of antibod for antigen
A e he e an c o eac ion
ha o ha e o con ide o
make he e mo e pecific?
Affinit & Avidit for pair
Ho ong i he binding
be een he Ag and he Fab
egion
O e all binding
Environmental conditions
Li ea e ide Na i e a al f m
Form of Antigen
e e ce
Degraded in C l E d c ic E d c ic
(e d ge ) e icle e icle
(e ge ) (e ge )
Sh i he Mecha i m b hich
Mac hage ca e , ce a d di la
Ag e ide MHC Cla II c m le f
ec g i i f Th1 cell
CD4 Ac i a ed CD4(+) T cell ecificall Th1 T CELL DELIVERS ITS LETHAL HIT ON TARGET CELL
Activation b e ca elea e c ki e ha ca
Cell i fec ed i h a i ha e eg la ed e e i f
ac i a e Mac hage ha b i g he e ic la
Fa
a h ge i hi i e d c ic e icle .
I l e Fa -Fa liga d ef i de a ge cell
C ki e f m Th1ca d ce i ic ide
Fa -Fa liga d
a d ge adical f killi g f a h ge .
Bi di g f Fa (membe f TNF famil )
Th1, Th2, Th17
f d i a ge cell
Diffe he c ki e d ced
Fa L f di Tc ic cell hich i
Th1 ec i me a d life a i
he Fa ece
f mac hage
Ac i a e a dea h d mai & a ge cell ill
Th2 IL-4, 5, 10 (i l i g B cell ;
be de ed ia a i
h m al imm i )
CTL i e ac i / Ag e ide-MHC ca e elea e f l ic
Th17 i l i g a h ge f d
g a le a d U eg la i f Fa L. E f g a le
i he m c al a ea
a ge cell l Fa -Fa L i e ac i i d ce a i
T CELL ACTIVATION, PROLIFERATION and Ac i a ed CD4(+) T cell ecificall Th1 b e ca elea e
DIFFERENTIATION of CD8 & CD4 T CELLS c ki e ha ca ac i a e Mac hage ha b i g he
e ic la a h ge i hi i e d c ic e icle . C ki e
f m Th1 ca d ce i ic ide a d ge adical f
killi g f a h ge .
Marker Function
Surface Ig A ige ecific ec g i i
MHC, class II A ige e e ai
(HLA-DR/DQ)
B7-1 & 2 T-cell ac i a i
B Cell Activation b T dependent Ag
CD40 Recei e ac i a i ig al f m T and B Cell Interaction
T cell
B cell ec g i e a d ca e he Ag h gh hei BCR a d
CD19-21 U i e B cell ma ke bi di g f Ag he ece lead i e ali a i f he Ag
Mitogen receptor Bi di g a d ac i a i b bac e ial ( ia ece media ed e d c i ).
d c The TD Ag i ce ed i e ide a cia ed i h M Cla
Ma e B cell ld ha e face Ig (IgM & IgD) II a d di la ed he face f ec g i i b CD4 hel e
Si ce B cell i al ca able f bei g a APC, MHC Cla II i T cell
al e e he face
e i helial)
Classical Path a
C1q 410 150 Bi d Fc egi f IgM a d IgG
C1r 85 50 Ac i a e C1
C1s 85 50 Clea e C4 a d C2
C4 205 300-600 Pa f C3 c e a e (C4b)
C2 102 25 Bi d C4b f m C3 c e a e
C3 190 1,200 Ke i e media e i all a h a
C5 190 80 I i ia e memb a e-a ack c m le
C6 110 45 Bi d C5b i MAC
C7 100 90 Bi d C5bC6 i MAC
C8 150 55 Sa e f ma i memb a e
C9 70 60 P l me i e ca e cell l i
Alternati e Path a
Factor B 93 200 Bi d C3b f m C3 c e a e
Factor D 24 2 Clea e Fac B
Properdin 55 15-25 S abili e C3bBb-C3 c e a e
MBL Path a
MBL 200-600 0.0002-10 Bi d ma e
MASP-1 93 1.5-12 U k
MASP-2 76 U k Clea e C4 a d C2
Fc f agme c alli able; Ig imm gl b li ; MAC memb a e-a ack c m le ; MBL ma e-bi di g lec i ; MASP MBL-a cia ed e i e ea e
HIGHLIGHTED IN RECORDING
Wi h ega d he c ce a i fc e e c e :
M ab da i e m C3
Im a ke i e media e i all a h a f he c m leme
Th ee a h a : cla ical, al e a i e, a d lec i a h a
C e ge ce i f all a h a
All a h a ill e e all mee i hi c m e
Im a marker of inflammation
A e i flamma di ea e ha he a ie i e e ie ci g
Sec d m ab da i e m C4
Im a a f C3 c e a e
E me d ced ac i a e C3
Wi h ega d he i e ec a eigh f c e e c e :
La ge C1q
Rec g i i i
The C1 c m le i c m ed f C1 , C1 , a d C1 .
Ve c cial a f he C1 c m le
I bi d he Fc i / ece f IgM a d IgG Ab ha ld i i ia e he a f he c m leme ac i a i ,e eciall i
he cla ical a h a
The other complement components are also necessar for complement acti ation.
c m le e ld be e g lfed di ed f b
hag c e .
Tagged b C3b (opsonin)
Whe la ma cell elea e Ab
F ll i g cla ical a h a ei he be de ed
b l i i ed b mac hage
SLE ( e ic e he a )
i flamma i & de ii f imm e c m le i
diffe e ga
Fig e 1. O i ai
LPS
Imm e
S im la e B cell e e a d Ab d ci Acti ating (bac e ial Ma eg
c m le e
Al e a i e Pa h a C3d i ec g i ed b CR2 substance ca le) mic bial cell
(IgG/IgM)
IgA
e e i B cell ; e ha ce B cell e e
mic be Recognition C3, Fac B, MBP, MASP-1,
C1q, C1 , C1
unit Fac D MASP-2
F he d ci &e e i f Ab
C3
C4b2a C2bBb C4b2a
con ertase
C5
C4b2a3b C3bBb3b C4b2a3b
con ertase
MAC C5b6789 C5b6789 C5b6789
RECOGNITION UNIT
Classical
C1 c m le
C1q bi d he Fc i f he Ab
C1r & C1s a e m ge ha ld e e all
ge e a e e me ac i i begi he ca cade
Alternati e
HISTORY OF COMPLEMENT C3
Ehrlich le f "c m leme i g" a ib die i de i g Fac B
mic ga i m . Fac D
Bordet el cida e le f C' Lectin
Q i e imila he cla ical b ld be i g a
diffe e e m
MBP imila C1
MASP-1 & MASP-2 imila C1 a d C1
Ba g ad g f c g a a d b O l a mall e ce age
f ag e f C2. f clea ed C3 m lec le
C2b ld ha e he bi l gic f c i ; bi d a ige ; m
g e he bl d eam. a e h d l ed b ae
C2a ld al be a ached he m lec le a d deca i
face f he a ge cell; emai he he fl id ha e
a ge cell. ★ O ce C3b i b d C4b2a, i
★ C2 ge e i cl el a cia ed ld he me ge i h C4b2a
i h he ge e f Fac B f m C4b2a3b (C5 con ertase).
(al e a i e ah a )
ch m me 6
C4b and C2a ( hich ha he highe MW), i he
e e ce f Mg2+, ld f m C4b2a ( i e a :
i dica e ha i i a ac i e e me)
he C3 con ertase.
C m le i e able
Half-life: 15 3 mi
C3 m be b d icl
3. C3 c e a e h d l e ma C3 m lec le . S me
c mbi e i h C3 c e a e f m C5 con ertase MEMBRANE ATTACK COMPLEX (MAC)
[RECALL]
C3 : m ab da i he bl d 4. The C3b c m e f C5 c e a e bi d C5, e mi i g
Maj a d ce al c i e f all f he C4b2a clea e C5
ah a he i al i . ( la ma C5 c e a e ill li he C5 i C5a a d C5b.
c c. 1 - 1.5 mg/mL) C5a g e he ci c la i a d ha e
ALL f he a h a ld e e all he bi l gic effec
mee i he C3. ( i al i ) C5b emai i he a ge cell/Ag;
I he e e ce f C3 c e a e, C3 ill be a ache cell memb a e ( a f MAC)
clea ed i C3a a d C3b. C5b ld e e all bi d C6
Clea age f C3 (MW: 190,000) C3b S li i g f C5 a d he clea age f C3 e e e
e e e he m ig ifica e i he he m ig ifica bi l gical c e e ce f he
e ie ce f c m leme ac i a i c m leme em
★ l e ide chai , al ha ( ) C5b e emel labile a d a idl
a d be a ( ) i ac i a ed le bi di g C6
chai c ai a
highl eac i e hi e e
g .
C3a he bi l gic f c i
★ em ed b clea age f a i gle
b d i he chai , he hi e e
i e ed
C3b highe MW; emai a ge cell
face; f c i a a ef l i
★ C3b i a im a e i he
ce f C ac i a i . 5. C5b bi d C6, i i ia i g he f ma i f he
★ C3 i ca able f bi di g membrane-attack comple .
h d l g O ce C5b bi d i h C6, b e e bi di g f
ca b h d a e a d ei i he e mi al c m e fC ld ha e
immedia e ici i I cl de bi di g f C7, C8, and C9
★ If C3b i b d he a ge The e C ld ha e e ma ic
cell/Ag, i ld ha e a e h ac i i ; he ld j bi d he C6.
life. (half-life: 60 mic ec d )
All e e i m ch malle am i
e m
★ C6 a d C7 (MW each: 110, 000):
imila e ie
★ C8 (MW: 150,000): 3 di imila
chai i di lfide b d
★ C9 (MW: 70,000): i gle
l e ide chai
Af e he bi di g, he memb a e a ack-c m le
(MAC) ill be f med.
C m ed f C5b-C9
The h d h bic a f he MAC ld
a ach/a ch he a ge cell m el i b
elea e f a e
Ca b e mi al e d i h d h bic
Ami - e mi al e d i h d hilic A ea a ce f MAC de he e ec c c e
Relea e f ae l f elec l e
ALTERNATIVE PATHWAY
ab b a g a ge ce ae e e
a aba g e ec e a ge cell a e O igi all called he properdin s stem beca e he ei
de ed e di a h gh be he mai i i ia f hi
The c m le f C5b-C6-C7-C8 and C9 i k ah a .
a C5b-9 or MAC P e di maj f c i i abili e a ke
If he c m le i soluble i ci c la i , i i k e me c m le f med al g he a h a a d
a sC5b-9. ha he he f m f ac i a i a e m e
Mea eme f he le el f C5b-9 i a mi e .
i dica f he am f e mi al Properdin fi h gh be a
a h a ac i a i ha i cc i g. im a c m e (i i ia ) f hi
Whe f med, he MAC e e a e f 70 ah a ,b a f d l be a
100 ha all i a i a d f he abili i g fac f he C3 c e a e
memb a e P ei properdin, a c i e f mal e m
I fl f ae a dl f elec l e de ci i h a c ce a i fa ima el 5 15
f a ge cell g/mL.
P e e ce f C9 g ea l eed he l i Fi de c ibed b Pilleme i he 1950 60
Ab e ce f C9 Ca be ac i a ed i ih a imm e c m le e
fficie e ba i f he I f ci mai l a a am lifica i l f ac i a i
memb a e ca cc i he a ed f m he cla ical lec i a h a
ab e ce f C9 I me ca e , he i i ial ac i a i i b he
Deficie cie i C9 a ea la gel cla ical a h a .
be ig Al e a i e a h a ca c i e
e ha ce he ac i a i .
Al h gh e di ha bee c fi med bi d a d
i i ia e ac i a i , he ima f c i f e di
i abili e he C3 c e a e f med f m
ac i a i f he fac .
Se m ei Fac B a d Fac D a e i e hi
ah a
C3 ke c m e
C m e : C3, factor B, factor D, properdi
T igge i g b a ce :
Mic bial d c
Pa h ge
C m e f bac e ia, f gi, aa ie
O e a ic la a a i e ha igge I ac a he eed f ac i a i f he
al e a i e a h a a me al e a i e a h a .
Bac e ial cell all (LPS) 2. Fac B bi d C3a, e e i e ac ed b fac D.
F gal cell all Clea age ge e a e C3bBb, hich ha C3 c e a e
Yea ac i i
Vi e Factor B imila C2 i he cla ical a h a
O he f eig b a ce he - a h ge F m a i eg al a f a C3 c e a e
A ib die (IgG, IgA) O ce Fac Bi b d C3b, i he e e ce f
Ve m Fac D, i ca be clea ed (b Fac D).
He e l g e h c e Factor D
P l me Pla ma ei ha g e h gh
Ca b h d a e a c f ma i al cha ge he i
Vi all i fec ed cell bi d Fac B.
T m cell li e Se i e ea e i h MW f
Se e a i e f bi di g he c m le C2bBb 24,000
( e f he e d d c ) C ce a i i la ma i he
C e i f C3 fi e l e f all c m leme ei
Na i e C3 able i la ma ( 2 g/mL)
Wa e i able h d l e a hi e e b d, h Clea e Fac B i iece :
a e l ac i a i g a mall mbe f he e Ba (MW: 33,000) a d Bb (MW:
m lec le 60,000)
TABLE 7-1 I i ia f he al e a i e a h a f c m leme ★ Bb emai a ached C3b
ac i a i f mi g he i i ial C3 c e a e
PATHOGENS AND PARTICLES OF MICROBIAL ORIGIN
f he al e a i e a h a .
★ Ra idl i ac i a ed le i
1. Ma ai f g am- ega i e bac e ia
2. Li l accha ide f m g am- ega i e bac e ia
bec me b d a ie e
3. Ma ai f g am- i i e bac e ia f he igge i g cell la a ige
4. Teich ic acid f m g am- i i e cell all
5. F gal a d ea cell all ( m a )
6. S me i e a d i i fec ed cell
7. S me m cell (Raji)
8. Pa a i e ( a me )
NONPATHOGENS
S c e f all h ee cla e f ec g i i
m lec le i imila ha f C1 all cla ed a
c llec i
e e ce f DAF h cell ec
hem f m b stander l sis ( ed i Regulation at assembl of membrane-attack comple (MAC)
di c imi a i f elf f m elf 1. S ei e e i e i f C5b67 MAC c m e i
beca e f eig cell d e
hi b a ce.) he memb a e
All f he e ac i c di a i i h he
Fac I
Factor I a ei e ea e ha i ac i a e C3b M im a maj eg la : S protein or
a d C4d eg la e C3 c e a e itronectin
C4BP- ab da i la ma S l ble c l ei ha i e ac ih
Ca able f c mbi i g i h ei he fl id C5b-7 c m le e e hi f m
ha e b d C4b; he ef e, C4b bi di g he cell memb a e
ca bi d C2 a d i made a ailable The c m le e MAC i f med d e
f deg ada i b Fac I. he S ei
If C4BP a ache cell-b d C4b, i ca P e e a ack f ea b cell
di cia e i f m C4b2a c m le e , Bi di g f
ca i g he ce a i f he cla ical C8 a d C9 ill ceed , b
ah a . l me i a i f C9 d e cc ;
he ef e, he c m le i able i e
3. I hibi -b d C4b i clea ed b fac I i elf i he cell memb a e d ce
4. I al e a i e a h a , CR1, MCP, or factor H e e l i
bi di g f C3b a d fac B 2. Homologous restriction factor (HRF) membrane
Factor H c cial i eg la i g C3 c e a e inhibitor of reacti e l sis (MIRL CD59) bi d C5b678,
Ac b bi di g C3b, e e i g he e e i g a embl f l -C9 a d bl cki g f ma i f
bi di g f Fac B. MAC
Ac a a c fac ha all Fac I
b eak d C3b.
I a ea ha l h e m lec le ih
igh l b d Fac H ac i e high-affi i
bi di g i e f Fac I
CD59 he memb a e i hibi f eac i e l i
I hibi -b d C3b i clea ed b fac I
(MIRL)
AFTER assembl of con ertase acti it Bl ck f ma i f MAC
C3 c e a e a e di cia ed b C4bBP, CR1, fac H, Pe e i he cell memb a e f all
a d deca -accele a i g fac (DAF) ci c la i g bl d cell
COMPLEMENT RECEPTORS
Al k a er throderma F C3 a d C4 a d he c m leme c m e
desquamati um Nephelometr ba ed mea eme
8. C5-C8 deficienc Ne e a i fec i , me i g c ccal f am ligh ca e ed
me i gi i , di emi a ed g heal di ea e The e ld be a ib die ecific
9. C7 deficienc Nei e ia i fec i , SLE, Ra a d' f C3 f ma i f imm e
he me , Scle de ma like d me, a c li i c m le mea ed h gh
10. DAF deficienc PNH CHINH deficie c : he edi a e hel me
a gi edema Rel he eci i a i f a ige
11. Factor D deficienc P ge ic i fec i (c mm ; ( he c m leme c m e bei g
beca e if a ie ha C2 deficie c , Fac D ill al be mea ed) a d a ib d .
deficie ) The m e a ige a ib d
c m le e ha a e e e , he
Table 7-4 Deficiencies of Complement Components m e a beam f ligh ill ca e a
i a e h gh he l i .
Deficient Component Associated Disease Turbidimetr
L -like d me Rel he eci i a i f a ige
C1 ( , , )
Rec e i fec i ( he c m leme c m e bei g
L -like d me mea ed) a d a ib d .
C2 Rec e i fec i RID (radial immunodiffusion)
A he cle i Me h d ha ill be ed i he lab
Se e e ec e i fec i f C3 de e mi a i
C3
Gl me l e h i i U e aga e gel i hich
C4 L -like d me Immunologic ecific a ib d i i c a ed.
assa s Se m e e a he a ige a d
C5 - C8 Ne e a i fec i i laced i ell ha a e c i he
C9 N k di ea e a cia i gel.
Diff i f he a ige f m he
C1-INH He edi a a gi edema
ell cc i a ci c la a e
DAF Pa mal c al hem gl bi ia ELISA f C1 i hibi (C1-INH i hibi )
MIRL Pa mal c al hem gl bi ia C m e f a da di ed g : C1 , C4,
C3, C5, Fac B, Fac H, a d Fac I
Fac H Fac I Rec e ge ic i fec i
N e f he a a f i di id al
P e m c ccal di ea e c m e a e able di i g i h he he
MBL Se i he m lec le a e f c i all ac i e
Ne e a i fec i
RID a d e hel me a e b h e i i e
P e di Ne e a i fec i e
MASP-2 P e m c ccal di ea e
C1-INH C1 i hibi ; DAF deca -accele a i g fac ; MASP-2
ma e-a cia ed e i e ea e; MBL ma e-bi di g lec i ; MIRL
memb a e i hibi f eac i e l i
N di ec di ea e a cia i i C9 deficie c beca e he
MAC ca ill be effec i e
Deficie c i MBL i al c mm
P e m ia, e i , e m c ccal di ea e i
i fa , ca ce , SLE Radial Immunodiffusion
The eci i i i g i mea ed
The la ge he i g, he highe he am f C3
LABORATORY DIAGNOSIS
Hemol tic Titration (CH50) assa - m c mm
Mea eme fc m e a a ige i e m
C de c ee i g e de ec de ec
de e mi e i le el (if deficie c i e e ) Classical
c m leme ac i i i e m
Mea eme f he f c i al ac i i path a
A deficie cie i he c m leme ,
U all d ei di a lab a ie ; d e i efe e ce assa s
e eciall i he cla ical a h a , ld
eciali ed lab a ie
lead ab mal e l .
C m e i e he cla ical
a h a h ld be i e iga ed
If both are lo , he e i m likel a blem i
he e mi al a h a (f ma i f he MAC)
Tha a h a i ha ed b b h he
cla ical a d al e a i e a h a
I e i ecime c llec i a d
ha dli g a e al c ide ed.
Al ible, if he e i fficie
ac i a i f c m leme h gh a e
a h a , ha he ac i a i c ld
c me e gh c m e l e he
f ci f he he a h a .
★ C m leme ac i a i i a el limi ed
j e ah a .
Sc ee i g e l ; if e l a e ab mal,
ceed a m e ecific e f C f c i
mea eme
T ical Sc ee i g Te f C' ab mali ie i l e :
C3, C4 a d hem l ic c e
Te i g f C3 C4a C5a Ba m i i g i flamma
ce e
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |1
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |2
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
● In a normal immune response, the activity of Th2 cells and Th1 cells are
balanced
○ Results in protective immunity that does not harm the host.
● In people with allergies, Th2 cells predominate.
○ Results in production of several cytokines, including IL-4 and IL-13.
ROLE OF IgE
ALLERGENS
● Common allergens are bee venom, pollen, drugs, mold spores, animal hair,
dander, and the feces and particles from mites.
● Other examples of common allergens include peanuts, eggs, and pollen.
○ Food allergens include cow’s milk, soy, wheat, fish, and shellfish.
● Allergens can be inhaled, ingested, or enter through the skin, mucus
membrane, digestive tract or genitourinary tract.
● Avoidance of allergen is the first line of defense against allergies.
Mast cell with IgE antibody on Fc receptor
ALLERGEN PROCESSING ● Binding of IgE to cell membranes increases the half-life of the antibody from
2 or 3 days to at least 10 days.
○ Once bound, IgE serves as an antigen receptor on mast cells and
● Key immunologic components involved are IgE antibody, mast cells,
basophils.
basophils, and eosinophils.
● Just like any other antigen, allergens are processed by macrophages and
presented to T Helper cells. IgE CROSS LINKING
● In allergic individuals a subclass of Helper T cells (TH2 cells) stimulates B
cells with specific receptors for allergen to form IgE antibodies. ● Activation phase
○ IgE is the least abundant in the circulation. ● When allergen is encountered again it is bound by specific IgE molecules on
○ Th2 cells regulate IgE. the surface of mast cells.
● IgE is primarily synthesized by B cells and plasma cells in the: ○ Leads to degranulation of mast cells.
○ Lymphoid tissue of the respiratory tract ○ Large numbers of these receptors are found on mast cells and
○ Lymphoid tissue of the gastrointestinal tract basophils.
○ Lymph nodes
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |3
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
● The allergen cross-links the IgE and this causes aggregation in the surface NEWLY SYNTHESIZED MEDIATORS
of the FcRI receptors to release the contents of its granules and to start
synthesizing new chemical mediators.
○ Initiates complex intracellular signaling events involving multiple ● When the mast cell is triggered to release the contents of its granules, it also
phosphorylation reactions, an influx of calcium, and secretion of starts to synthesize and release other mediators (secondary mediators) -
cytokines. leukotrienes, bradykinin, prostaglandins, and platelet-activating factors.
■ Increase in intracellular calcium triggers rapid ● Secondary mediators are more potent than the primary mediators and are
degranulation of the mast cells and basophils. responsible for a late-phase allergic reaction
○ Seen in some individuals 6 to 8 hours after exposure to the antigen.
○ Eosinophils play an important role in the late-phase reaction.
MAST CELL DEGRANULATION ● Eosinophils , neutrophils, Th2 cells, mast cells, basophils, and
macrophages, exit the circulation and infiltrate the allergen- filled tissue.
● Mast cell granules contain preformed mediators (primary mediators) such as ○ Release additional mediators that prolong the hyperactivity and
histamine, serotonin and eosinophil, and neutrophil chemotactic factors may lead to tissue damage.
(ECF and NCF). ● These molecules also result in dilation of small vessels (vasodilation),
○ Chemotactic factors will recruit eosinophils and neutrophils in the leakage of fluid (edema), smooth muscle contraction, increased mucus
area = upsurge in eosinophils and neutrophils. secretions, and pain.
○ Histamine is the most important and abundant chemotactic factor.
■ Primarily increases permeability of blood vessels and
usually promotes contraction of smooth muscles like
trachea (contraction of trachea = breathing difficulties).
○ The most abundant preformed mediator is histamine (10% of total
weight of the granules in mast cells).
○ Other primary mediators include heparin, eosinophil chemotactic
factor of anaphylaxis (ECF-A), and proteases.
● The release of preformed mediators is responsible for the early-phase
symptoms seen in allergic reactions.
○ Occurs within 30 to 60 minutes after exposure to the allergen.
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |4
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |5
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
○ Shifts the immune response to Th1-type of response and induces ○ Uses radiolabeled IgE to compete with patient IgE for binding sites
the development of T regulatory cells (Tregs) that release IL-10. on a solid phase coated with anti-IgE.
○ The IgG blocking antibodies mock-up the allergens before it ○ Due to expense and difficulty of use, RIST has largely been
reaches the IgE antibodies on mast cells. replaced by noncompetitive solid-phase immunoassays or
○ It is also suggested that hyposensitization therapy inhibits Th2 cells, nephelometry assays with enhanced sensitivity.
favoring IgG synthesis over IgE production. ● Chemiluminescent Enzyme Immunoassay
○ Automated immunoassays with high level of specificity and
TREATMENT sensitivity.
○ Can be run with a single allergen or as a multi-allergen screen using
a panel of allergens in a single run.
● Persistent asthma → leukotriene receptor antagonists and mast cell ● Protein Microarray
stabilizers ○ Allows for parallel detection of IgE antibodies to more than 100
● Corticosteroids potential allergens using only 20 μL of patient serum.
○ Block recruitment of inflammatory cells and their ability to cause ■ Serum is incubated with a biochip containing miniature
tissue damage. spots to which the purified allergenic components have
● Systemic anaphylaxis → epinephrine injection been applied.
● Omalizumab (anti-IgE monoclonal antibody) ■ If allergen-specific IgE is present, it will bind to the
○ Composed of human IgG framework genes recombined with appropriate spots.
complementarity-determining region genes from mouse ■ The chip is scanned by a laser for fluorescence following
anti-human IgE. addition of a fluorescent-labeled anti-IgE.
○ Prevents circulating IgE from binding to mast cells and basophils ○ Highly sensitive and specific
and sensitizing them by binding to the Cε3 domain of human IgE, ● Immunocap
which is the binding site of FcεRI receptors.
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |6
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
○ This is due to the binding of antigen. ■ Rh, Kell, and Duffy antigens may also be involved in
immediate transfusion reactions.
● Hemolytic disease of the newborn (Rh disease)
● Autoimmune hemolytic anemia
○ Warm Autoimmune Hemolytic Anemia
■ Referred to as idiopathic autoimmune hemolytic anemia
■ Associated with IM, CMV, chronic active hepatitis, chronic
lymphocytic leukemia and lymphomas.
○ Drugs that can induce antibody production which causes hemolytic
anemia:
■ Penicillins and cephalosporins
■ Quinidine and phenacetin
■ Methyldopa
○ Paroxysmal Cold Hemoglobinuria
MECHANISMS ■ Occurs after infection with measles, mumps, chickenpox
or IM.
● Autoimmune thrombocytopenia
● Antibody-dependent, complement mediated cytotoxic reactions
● Myasthenia gravis
○ Involves the activation of the classical pathway of the Complement.
○ Receptors of acetylcholine are blocked, leading to muscle
○ Lead to the formation of MAC and later cell lysis.
weakness.
● Antibody-dependent, cell-mediated cytotoxicity antibody
● Pemphigus
○ Coated cells are lysed by effector cells, such as natural killer (NK)
● Pemphigoid
cells and macrophages.
● Goodpasture’s syndrome
■ Mediated through binding of IgG antibody to its
○ Production of antibodies that reacts with basement membrane
corresponding antigen on the target cell and to Fc
protein.
receptors on macrophages or natural killer cells.
■ Usually affects glomeruli in the kidney and pulmonary
○ Coating of the cell surface by antibodies promotes opsonization,
alveolar membranes.
and subsequent litho phagocytosis.
● Grave’s disease
■ Opsonization occurs either through binding of IgG
● Insulin-dependent diabetes (Juvenile diabetes)
antibody to Fc receptors on macrophages and neutrophils
or binding of cell surface C3b to complement receptors on
phagocytic cells.
● Antireceptor antibodies
○ Autoimmune hypersensitivity against solid tissue, Hyperacute graft
rejection
○ Cell damage will result from mechanisms of the antibodies and
antibody-dependent through binding of the IgG to the
corresponding antigen on the target cell.
● Transfusion reactions Normal individuals may have autoantibodies in their serum, but it is not a diagnosis
○ ABO, Rh, Kell, Kidd and Duffy. of the disease unless these autoantibodies block receptors and lead to different
○ Antigens most involved in delayed reactions include those in the conditions.
Rh, Kell, Duffy, and Kidd blood groups.
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |7
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |8
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
● The complement components, C3a and C5a also act as chemotaxins and
attract polymorphonuclear granulocytes (PMN) and macrophages to the site.
● The complexes deposit on the vessel wall lead to vasculitis and the PMN
tries to destroy them by releasing enzymes, damaging the wall.
● Platelet aggregation causes microthrombi (small clots) which interfere with
the local blood supply.
● The process may resolve or result in chronic inflammation.
● Autoimmune disease
● Hypersensitivity pneumonia
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |9
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
● Infection LABORATORY EVALUATIONS
● Drug reactions
● Indirect Immunofluorescence
DISEASE ASSOCIATED WITH IMMUNE COMPLEXES ● EIA
TYPE EXAMPLES ● FLuorescent Microsphere Multiplex Immunoassay
● Latex Agglutination
Rheumatoid arthritis, systemic lupus erythematosus, ● Nephelometry
Autoimmune disease Sjogren’s syndrome, mixed connective tissue disease, ● Measuring complement levels
systemic sclerosis, glomerulonephritis
Neoplastic disease Solid and lymphoid tumors TYPE IV - CELL MEDIATED HYPERSENSITIVITY
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |10
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
■ Plants (eg. poison ivy, poison oak), metals (eg. nickel ■ Cosmetics
salts), and components of hair dyes and cosmetics. ■ Medications (topical anesthetics, antiseptics, and
● Examples: antibiotics)
○ Allergy or infection ■ Latex
○ Contact dermatitis ○ Produces a skin eruption characterized by erythema, swelling, and
○ Tuberculin reaction the formation of papules that appears from 6 hours to several days
after the exposure.
TYPE IV HYPERSENSITIVITY REACTIONS ARE MEDIATED BY ■ Papules may become vesicular with blistering, peeling,
ANTIGEN-SPECIFIC EFFECTOR T CELLS and weeping.
● Patch test
SYNDROME ANTIGEN CONSEQUENCE ○ Gold standard in testing for contact dermatitis.
○ Done when the patient is free of symptoms or at least has a clear
● Proteins: ● Local skin swelling:
test site.
○ Insect venom ○ Erythema
Delayed-type ○ A nonabsorbent adhesive patch containing the suspected contact
○ Mycobacterial ○ Induration
hypersensitivity allergen is applied on the patient’s back and the skin is checked for
proteins (tuberculin, ○ Cellular infiltrate
a reaction over the next 48 hours.
lepromin) ○ Dermatitis
○ Positive Test - Redness with papules or tiny blisters
● Haptens: ● Local epidermal reaction: ■ Individuals with deficient cell-mediated immunity will
○ Pentacecacatechol ○ Erythema display anergy -- absence of positive reactions for all of
(poison ivy) ○ Cellular infiltrate the common antigens used in the skin test.
Contact ○ Dinitrofluorobenzene ○ Vesicles ○ Final evaluation is conducted at 96 to 120 hours.
hypersensitivity (DNFB) ○ Intraepidermal ● Involves antigen-sensitized T cells or particles that remain phagocytized in a
● Small metal ions: abscesses macrophage and are encountered by previously activated T cells for a
○ Nickel second or subsequent time.
○ Chromate
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |11
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
1ST EXPOSURE
● TDH that bear matching antigen receptors come in contact with the antigen
and proliferate clones, which sensitize the individuals.
● 2nd exposure activates TD cells which proliferates and release lymphokines Cells Involved Mast cells, Effector cells Macrophages, Antigen -
○ It will stimulate macrophage to attack targeted epithelial cells basophils, (macrophages, mast cells specific T cells
granules PMN
(histamine) leukocytes)
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |12
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
Description Mediated, dependent; complex mediated, mast cells and antibody and complement cytokines that
immediate complement or mediated delayed type basophils complement, proteins. recruit
cell mediated (immune opsonization Neutrophils are macrophages
complex , or ADCC recruited and and induce
disease) ● Cell function release inflammation
inhibited by lysosomal or activate
Mechanism of Allergic and Target cell Immune Inflammation,
antibody enzymes that cytotoxic T
Tissue Injury anaphylactic lysis; cell complex cellular
binding cause tissue cells to cause
reactions mediated deposition, infiltration
● Cell function damage. direct cell
cytotoxicity inflammation
stimulated damage.
Examples ● Anaphylaxis ● Transfusion ● Arthus ● Allergy or by antibody
● Hay fever reactions reaction infection binding
● Allergic ● Autoimmune ● Serum ● Contact
rhinitis Hemolytic sickness dermatitis
TYPE OF PATHOLOGIC IMMUNE MECHANISMS OF
● Asthma Anemia ● SLE ● Tuberculin
HYPERSENSITIVITY MECHANISMS TISSUE INJURY &
● Food allergy ● HDN ● Rheumatoid and anergy
DISEASE
● Urticaria ● Drug arthritis skin tests
reactions ● Drug ● Hypersensiti Immediate Th2 cells, IgE antibody, mast ● Mast cell derived
● Myasthenia reactions vity Hypersensitivity cells, eosinophils mediators (vasoactive
gravis pneumonitis (Type I) amines, lipid
● Goodpasture mediators, cytokines)
’s syndrome ● Cytokine-mediated
● Grave’s inflammation
disease (eosinophils,
● Thrombocyt neutrophils)
openia
Antibody-mediated IgM, IgG antibodies against cell ● Complement and Fc
diseases (Type II) surface or extracellular matrix receptor-mediated
COMPARISON OF HYPERSENSITIVITY REACTIONS antigens recruitment and
activation of leukocytes
TYPE I TYPE II TYPE III TYPE IV
(neutrophils,
Immune macrophages)
IgE IgG or IgM IgG or IgM T cells ● Opsonization and
Mediators
phagocytosis of cells
Synonym Anaphylactic Antibody Complex Cell mediated ● Abnormalities in
mediated mediated or delayed cellular function (eg.
cytotoxic type hormone receptor
Timing Immediate Immediate Immediate Delayed signalling)
Antigen Heterologous Cell surface; Soluble; Autologous or Immune complex - Immune complexes of ● Complement and Fc
autologous or autologous or heterologous mediated diseases circulating antigens and IgM or receptor-mediated
heterologous heterologous (Type III) IgG antibodies deposited in recruitment and
vascular basement membrane activation of leukocytes
Immune Release of ● Cell Ag-Ab Antigen
Mechanism mediators from destruction complexes sensitized Th1
Ige sensitized caused by activate cells release
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |13
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
TYPE V HYPERSENSITIVITY
POST-TEST
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |14
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
○ Self-tolerance- ability of the immune system to accept self-antigens
UNIT 9B: IMMUNOPATHOLOGY: AUTOIMMUNE DISEASE and not initiate a response against them.
I. Autoimmunity ■ Can be caused by several mechanisms, e.g. Clonal
a. Concepts of Tolerance deletion, clonal anergy, suppression.
b. Immune Privilege ○ Tolerance to self antigens must be specific for the inducing epitope.
c. Mechanisms of Autoimmune Disorders
II. Autoimmune Disorders
a. Systemic Lupus Erythematosus CENTRAL TOLERANCE
b. Rheumatoid Arthritis
c. Hashimoto’s Thyroiditis
d. Grave’s Disease ● Destruction of self-reactive lymphocytes in the primary lymphoid organs.
e. Type 1 Diabetes Mellitus ● Early lymphocytes that are destroyed or undergo apoptosis.
f. Multiple Sclerosis ● Remember the bone marrow and the thymus because these are your
g. Myasthenia Gravis
h. Other Autoimmune Diseases primary lymphoid organs and this is where the B and T cells mature.
Sources:
PPT PERIPHERAL TOLERANCE
Video Part 1
Video Part 2
● Tolerance induced in mature lymphocytes from the outside of the primary
lymphoid organs.
AUTOIMMUNITY
CENTRAL TOLERANCE
● Autoimmunity – a condition in which the body responds to one or more
self-antigens (auto-antigens) ● Occurs in central or primary lymphoid organs, the thymus and bone marrow.
○ A condition by which there is a loss of self-tolerance. ● Negative selection
○ Self-tolerance is the normal characteristic/state of the immune ○ A process where T cells that express T cell receptors with a strong
system wherein the body can recognize or discern whether the affinity for these self-antigens are deleted by apoptosis.
antigen is self or non-self. ○ Occurs with both the immature, double-positive CD4+/CD8+ cells
● Horror autotoxicus – coined by Paul Ehrlich in the cortex and with the more mature, single-positive CD4+ or
○ Mechanism of Autoimmune Diseases CD8+ cells in the medulla.
○ “Horror” talks about how autoimmune disorders may inflict severe
outcomes to the patient. MECHANISMS
● Autoimmune Disease
○ Disorders in which immune responses are targeted toward
self-antigens and result in damage to organs and tissues in the 1. Clonal Deletion
body. ● Happens when early CD4+ T cells, which contain receptors for
○ These harmful effects may be caused by T-cell mediated immune endogenous molecules, are deleted after contact with self-antigens
responses or autoantibodies that are directed against host in the thymus.
antigens. ● Early B cells with self-reactive receptors are eliminated after an
interaction with self-antigens in the bone marrow.
CONCEPTS OF TOLERANCE
● Tolerance
○ Refers to the state of the immune system by which it is
unresponsive to an antigen.
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |15
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
■ Explains the interaction of the BCR or TCR with
its auto antigen which determines
survival/apoptosis.
○ BCR-Specific for non self antigen
■ BCR has been edited to not react with the self
antigen.
○ BCR-Specific for self antigen
■ There is affinity towards the self antigens which
will undergo apoptosis.
PERIPHERAL TOLERANCE
MECHANISMS
1. Anergy
● B-cell anergy
○ Anergic B cells cannot compete with the non-anergic
B-cells entering the B-cell follicles in the lymph
node/spleen → Apoptosis.
● T-cell anergy
○ Absence of co-stimulation the T-cell-APC interaction will
anergize the T-cell.
3. Receptor Editing
● Important mechanism of Central Tolerance for B-cell in which the
immature B-cell with BCR specific for auto-antigen is replaced with
a BCR specific for a non-self antigen.
○ Receptor avidity
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |16
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
ORAL TOLERANCE
IMMUNE PRIVILEGE
2. Fas-mediated activation causing cell death
● Fas mediated apoptosis ● Immune privileged sites
○ The removal of mature autoreactive B- and T-cells ○ Body sites that are usually unresponsive to pathogens, tumor cells,
● Fas (expressed by activated cells including T- and B-cells) binds to or tissue transplants.
FasL/Fas ligand present on several activated cell types. ○ Ex: cornea, testis, brain, placenta, and ovary
○ Once bound, Fas initiates apoptosis in the cell with Fas by 1. Lack of lymphatic drainage
activating the death domain. 2. Presence of blood barriers
○ Ex. Self-reactive B cells with Fas are susceptible to 3. Presence of immunosuppressive cytokines
Fas-mediated apoptosis if there is an interaction with Th ● E.g. IL-10,
expressing FasL. 4. Expression of FasL
○ If the FasL and Fas is bound together in the T or B-cell, it
will induce certain cascades inside the activated T or MECHANISMS OF AUTOIMMUNE DISORDERS
B-cell. e.g Death domain.
3. Induction of Treg
● Treg/Regulatory T cells/ Suppressor T cells ● Inability to maintain central and peripheral tolerance leads to the activation
○ They have the ability to down regulate the T-cell functions. of autoreactivity of immune system.
○ Purpose of T cells ● Initiation of autoimmune disorders:
■ “We do not want that CD4+ t helper cell to ○ Genetic
continuously activate because continuous ○ Environmental
activation of T cell will further activate several
cascades of the immune system, such as causing GENETIC PREDISPOSITION
reduction of interleukins or cytokines that can
also activate other mechanisms like phagocytosis
● No individual gene is sufficient to induce autoimmune disorders BUT the
etc”.
products of SEVERAL GENES are likely to induce one.
● Suppressive functions:
● Overexpression or underexpression of genes needed in:
○ Treg cells respond to signals via CTLA-4
○ Apoptosis, cellular survival, cytokine expression, BCR or TCR
○ Activation of LAG-3 which binds to MHC class II molecule
signaling, co-stimulatory molecule interactions, and immune
○ Secretion of IL-10 and TGF-B
clearance of apoptotic cells and immune complexes = autoimmune
phenotype.
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |17
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
● There is an association between the presence of certain human leukocyte HLA-DR3 Weak
antigen (HLA) types and the risk of developing a particular autoimmune Myasthenia gravis
HLA-B8 (class I) Intermediate
disorder.
○ The strongest link found is between the HLA-B27 allele and the Goodpasture’s Syndrome HLA-DR2 Strong
development of ankylosing spondylitis, an autoinflammatory
disease that affects the spine. ● If certain HLA products are mutated or if there are products that are
● Differences in MHC genes are thought to influence influence the affecting the HLAs, some of it will have disease correlation with autoimmune
development of autoimmune disease because the specific structure of the disorders.
MHC molecule can determine whether or not a self-antigen can attach to
the peptide-binding cleft of the molecule and subsequently be processed
and presented to T cells. ENVIRONMENTAL SUSCEPTIBILITY
● Polymorphisms in non-MHC genes can also be associated with
development of autoimmune disease. RELEASE OF SEQUESTERED ANTIGENS
○ PTPN22 gene- T- and B-cell receptor signaling
○ IL2RA gene- T-cell activation and maintenance of Tregs
○ CTLA4 gene- inhibitory effect on T-cell activation ● Sequestered/protected antigens may be found in privileged sites.
○ BLK gene- B-cell activation and development ● “Hidden’’ (cryptic) antigenic determinants that were unavailable when fetal
○ AIRE (autoimmune regulator) gene- development of T-cell tolerance clonal deletion was occurring can become available following
in the thymus. injury/infection.
● Single-gene mutations that can be inherited in a Mendelian fashion have ● Immunologic ignorance.
been associated with rare autoimmune disorders. ○ Production of autoantibodies to the lens of the eye following an
ocular injury.
○ Autoantibodies to sperm after a vasectomy.
HLA STRENGTH OF ○ Autoantibodies to DNA following damage to skin cells by
AUTOIMMUNE DISEASE
ASSOCIATION ASSOCIATION overexposure to UV rays from the sun.
MHC CLASS I
MOLECULAR MIMICRY
Ankylosing spondylitis
HLA-B27 Strong
Reiter’s Syndrome ● Refers to the fact that many bacterial or viral agents contain antigens that
closely resemble the structure or amino acid sequence of self-antigens.
Grave’s Disease HLA-B8 Weak
● Exposure to foreign antigens may trigger immune responses that
Psoriasis vulgaris HLA-Cw6 Intermediate cross-react with similar self-antigens.
● Autoreactive B cells can be activated, synthesizing antibodies if T-cell
MHC CLASS II recognizes a foreign antigen with an epitope that can cross-react with
Rheumatoid Arthritis HLA-DR4 Strong self-antigen.
● Examples:
Sjögren’s Syndrome HLA-DR3 Weak ○ M protein of S. pyogenes, which cross-react with sarcolemmal
HLA-DR2 Intermediate heart muscle (cardiac myosin) = seen in Rheumatic fever.
SLE ○ A peptide derived from Coxsackievirus cross reacts with a peptide
HLA-DR3 Weak
from glutamic acid decarboxylase (an antigen in Beta-islet cells).
Celiac Disease HLA-DR3 Strong This is recognized by T cells which is seen DM type 1.
HLA-DR3 Intermediate
DM Type 1 POLYCLONAL ACTIVATION
HLA-DR4 Strong
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |18
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
● Microbial antigens (ex. LPS/LTA) may nonspecifically activate B cells or T Platelet membrane protein;
cells, some of which may render these cells to become autoreactive = ITP B cells/autoantibody
Integrin
autoimmune disease.
Salivary duct antigens; SS-A,
Sjögren’s Syndrome B cells/autoantibody
SS-B
DRUG AND HORMONAL CAUSES
Centromeric proteins of
● Quinidine, sulfathiazole are adsorbed onto an endogenous molecule/cell Scleroderma fibroblasts; Nucleolar antigens; Unknown
(e.g. platelets) = formation of antigenic hapten-carrier complex. Scl-70
○ Antibody to the drug is formed and reacts with the drug on the
Pemphigus vulgaris Desmolglein 3 B cells; autoantibody
platelet membrane. Complement is activated, resulting in platelet
lysis. Renal and lung basement
Goodpasture’s syndrome B cells; autoantibody
● SLE-associated drugs → chlorpromazine, hydralazine and procainamide membrane collagen type IV
○ They have been observed to produce antinuclear antibodies (ANE)
in SLE. Thyroglobulin, Microsomal
CD4+ T cells; B
● Drug-induced HA → penicillin Hashimoto’s thyroiditis antigens and thyroid
cells/autoantibody
● SLE risk of women : men (10:1). peroxidase
○ Estrogen effects among females with SLE. MBP, Myelin oligodendrocyte
● Women are 2.7 times more likely to acquire an autoimmune disease than Multiple sclerosis CD4+ T cells; B cells
protein
men.
○ Women develop autoimmunity at an earlier age and have higher CD4+ T cells; CTLs; B
DM Type 1 Pancreatic B-islet antigen
risk for acquiring more than one autoimmune disease as compared cells/autoantibody
with men.
IgG; Citrullinated and CD4+ T cells; CTLs; B
○ Females have been found to have higher absolute CD4+ T-cell Rheumatoid arthritis
carbamylated proteins cells/autoantibody
counts and higher levels of circulating antibodies than men.
○ The stimulatory effects of female hormones may place women at CD4+ T cells (Th1 and
Psoriasis Unknown
greater risk for developing autoimmune disease. Th17)
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |19
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |20
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
● Diagnosis of SLE – SLICC Criteria (American College of Rheumatology, nuclear antigen
2012): (RNA component)
Chronic cutaneous lupus (discoid rash, mucosal High anti-dsDNA antibody Phosphoprotein
SLE, Sjögren’s
lupus, etc.) concentration Anti-SS-B(La) complexed to Finely Speckled
Syndrome, others
RNA polymerase
Oral or nasal ulcers Presence of anti-Sm
RNA polymerase, Homogenous staining of SLE, systemic
Nonscarring alopecia Positive APA Anti-nucleolar
nucleolar protein nucleolus sclerosis
Low complement (C3, C4,
Synovitis in > 2 joints Systemic
CH50) DNA
Anti-Scl-70 Atypical Speckled sclerosis,
topoisomerase I
Serositis Direct CoombsTest Scleroderma
Drug-induced
Different classes
Anti-histone Homogenous SLE, other
of histones
diseases
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |21
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
○ Multiplex bead assays
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |22
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
■ Less coarser and smaller compared to speckled.
■ Prominent staining at the nucleoli of interphase cells.
■ Staining of nucleoli varies on the number of nuclei present
in each cell.
■ Related with anti-RNP and anti-RNA.
■ associated with Scleroderma, Sjögren’s Syndrome and
MCTD.
○ Discrete/Centromere
■ Numerous discrete speckles in the nuclei of interphase
cells and chromatin of dividing cells.
■ Correlated with anti-centromere, anti-RNA, and anti-ENA.
■ associated with CREST Syndrome (Calcinosis, Reynauds,
Esophageal dysmotility, Sclerodactyly, Telangiectasia).
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |23
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |24
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
○ Sediment demonstrated in the same urine after centrifugation (right) ○ Radiographic evidence of erosions in the joints of the hands, the
● Urinary cast: wrists, or both.
○ [B] Hyaline
○ [C] Red cell
RHEUMATOID ARTHRITIS
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |25
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
LABORATORY DIAGNOSIS
IMMUNOLOGIC FINDINGS
● RF
○ Group of immunoglobulins IgG or IgM (typically an IgM) that
interacts specifically with the Fc portion of an IgG (target molecule)
● Sheep Cell Agglutination Test
○ (+) agglutination = (+) RF factor
○ Uses IgG as an agglutinin
● RF Latex Slide Test
● Rose Waaler Hemagglutination Test
● Nephelometric assays test for two other antibody classes of RF, IgG or IgA
TREATMENT FOR RA
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |26
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
● Patients develop a combination of goiter (or enlarged thyroid, it is irregular
and rubbery), HYPOthyroidism (symptoms: dry skin, decreased sweating,
puffy face with edematous eyelids, pallor with yellow tinge, weight gain,
fatigue, dry and brittle hair), and thyroid autoantibodies.
○ Thyroid autoantibodies: antithyroglobulin, antithyroid peroxidase
(microsomal antigen), and second colloid antigen (CA-2)
○ Immune destruction of the thyroid gland occurs.
● The thyroid shows hyperplasia with an increased number of lymphocytes.
○ Hyperplasia will cause goiter.
● Cellular types present: activated T and B cells (with T cells predominating),
macrophages, and plasma cells.
● Certain drugs can trigger autoimmune thyroiditis: denileukin diftitox,
interferon-alpha, IL 2, lithium, tyrosine kinase inhibitors
● Mild dry mouth (xerostomia) or dry eyes (keratoconjunctivitis sicca) related
to Sjögren syndrome.
● Pathology to the thyroid gland is mediated primarily by CD8+ cytotoxic cells
which bind to the thyroid cells and destroy them by releasing enzymes that
cause apoptosis or necrosis.
● Immune response results in the development of germinal centers that
almost completely replace the normal glandular architecture of the thyroid
and progressively destroy the thyroid gland.
LABORATORY DIAGNOSIS
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |27
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
● The most common cause of hyperthyroidism. ○ Binding assays (automated solid-phase ELISA & chemiluminescent
CLINICAL SIGNS immunoassays): a labeled TRAb reagent competes with the patient
antibody for TSH receptor bound to a solid phase.
○ BIoassays: difficult to perform because it requires tissue culture.
● Thyrotoxicosis, with a diffusely enlarged (hyperplasia) goiter that is soft
instead of rubbery.
● Symptoms TYPE 1 DIABETES MELLITUS
○ Nervousness, insomnia, depression, weight loss, heat intolerance,
sweating, rapid heartbeat, palpitations, breathlessness, fatigue, ● A chronic autoimmune disease that occurs in a genetically susceptible
cardiac dysrhythmias, and restlessness individual as a result of environmental factors.
● Result in elevated free and total T3 and T4; and decreased serum thyroid ● It is characterized by insufficient insulin production caused by selective
stimulating hormone (TSH) destruction of the beta cells of the pancreas.
○ Exophthalmos - hypertrophy of the eye muscles and increased ● Autoantibodies and CTLs are reactive against the pancreatic beta cells =
connective tissue in the orbit cause the eyeball to bulge out so that atrophy and fibrosis.
the patient has a large-eyed staring expression. ○ Beta cells are located in the pancreas in clusters called the islets of
■ There is conjunctival and periorbital edema which causes Langerhans.
the “bulging eyes” look. ● Progressive inflammation of the islets of Langerhans in the pancreas leads
to fibrosis and destruction of most beta cells.
● Diabetics carry HLA-DR2 or DR4 gene = increased risk for DM type 1
● It may also occur in the HLA-DQ region, especially in the coding of the DQ
chain.
● It is apparent that autoantibody production precedes the development of
T1D by months or several years.
● Thyroid-stimulating hormone receptor antibodies (TRAbs) ● Environmental influences include the possibility of viral infections and early
○ Major antibodies involved in Graves disease’ pathogenesis. exposure to cow’s milk.
○ When TRAbs bind to TSH receptors, they mimic the action of TSH ○ Linked viruses: rubella virus, cytomegalovirus, and Coxsackie B4
resulting in uncontrolled receptor stimulation with excessive release virus with diabetes, but most research is inconclusive.
of thyroid hormones. ○ Viruses can trigger autoantibody production by molecular mimicry.
TREATMENT TREATMENT
● US: first line of treatment involves radioactive iodine administered for 1-2 ● Daily injectable insulin has been the mainstay therapy for T1D.
years and results in a 30-50% long-term remission rate. ● Transplantation of pancreatic beta islet cells has been used for T1D patients
● Europe and Japan: patient is first placed on antithyroid medications with who have poor glucose control but this requires continual
beta blockers as adjuvant therapy then it is continued with drug treatment, immunosuppressive therapy in order to prevent rejection and the number of
radioactive iodine therapy, or surgery to remove part of the thyroid. suitable donors is limited.
● Surgery is recommended for patients resistant to drug treatment, but can
damage the laryngeal nerves and cause permanent hoarseness. LABORATORY DIAGNOSIS
LABORATORY DIAGNOSIS ● When T1D is suspected, tests for antibodies to GAD and IA-2A can be done
to confirm the diagnosis.
● Low/ undetectable levels of TSH and increased levels of FT4 ● Combined screening for IA-2A, ICA, and GAD antibodies appears to have
● TRAbs are highly indicative and one component of the diagnostic criteria of the most sensitivity and best positive predictive value for T1D in high risk
Graves disease. populations.
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |28
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
● An inflammatory autoimmune disorder of the central nervous system. ● Neuromuscular disease in which the nerve muscles do not function
● It is characterized by the formation of lesions called plaques in the white normally.
matter of the brain and spinal cord, resulting in the progressive destruction ● (+) antibodies to Acetylcholine receptors at the motor end plate = Blocking
of the myelin sheath of axons. the nerve impulses hence, damaging the neurons
● Considered a chronic progressive inflammatory disease with demyelination ○ Sensitivity of 80–90% for the diagnosis of myasthenia gravis.
of the nerves ● Other immunologic findings: serum antibodies to muscle-specific tyrosine
● Most closely associated with inheritance of particular HLA molecule coding kinase (MuSK)*.
for the beta chain of the DR subregion, namely DRB1*1501. ● Patients present with ptosis (drooping of eyelids), diplopia (double vision),
● Environmental factors: reduced exposure to sunlight, vitamin D deficiency, difficulty in chewing or swallowing, respiratory difficulties, limb weakness,
and cigarette smoking inability to retract the corners of the mouth often resulting in a snarling
● Associated viral infections: Epstein-Barr virus and Human Herpes 6 appearance, or some combination of these problems.
● Increased IgG in the CSF ● Characterized by weakness and fatigability of skeletal muscles.
● Early onset MG (EOMG) occurs before the age of 40 and affects
LABORATORY DIAGNOSIS predominantly females
○ Has strong association with the HLA haplotype, A1, B8 and DR3.
● Late onset MG (LOMG) occurs after 40 and is seen more often in males.
● IgG index = used to differentiates true IgG increase due to production rather ○ HLA Antigens B7 and DR2
THAN the increase in permeability of the BBB
○ IgG index = (IgG CSF/ Albumin CSF) / (IgG serum/ Albumin serum)
■ Ref range: 0.0 – 0.77 TREATMENT
○ CSF Electrophoresis = (+) Oligoclonal bands in the CSF
■ NON SPECIFIC, also seen in SLE, viral meningitis, ● Anticholinesterase agents to prevent destruction of neurotransmitter,
neurosyphilis acetylcholine, are used as main therapy
● Thymectomy should be performed on patients with thymoma.
CLINICAL SIGNS ● If these treatments aren’t effective, immunosuppression is recommended
● Treatment generally begins with high doses of corticosteroid drugs followed
by other immunosuppressive drugs (azathioprine or mycophenolate mofetil)
● Damage to tissue of CNS can cause visual disturbances, weakness/ to maintain response.
diminished dexterity in one or more limbs, locomotor incoordination,
dizziness, facial palsy, and sensory abnormalities (tingling/ “pins and
needles” that run down the spine or extremities and flashes of light seen on LABORATORY DIAGNOSIS
eye movement)
● Begins in young and middle-aged adults between ages 20 and 50. ● Radioimmunoprecipitation assay (RIPA): most commonly used procedure for
● Most patients with MS eventually develop progressive deterioration of the antibody to the ACHR, which is based on precipitation of the patient’s
CNS and functional disability. antibody with ACHRs isolated from human muscle.
● Immunofluorescence cell-based assays: patient serum is incubated with
TREATMENT HEK293 cells expressing all four ACHR subunits
○ Highly sensitive assay
○ Can detect antibodies directed toward ACHR clusters in patients
● Aimed at easing recovery from acute attacks and reducing the risk of future that were previously classified as seronegative by RIPA.
relapses. ● Fluorescence immunoprecipitation assays: uses ACHR subunits or MuSK
● Acute exacerbations are treated with corticosteroids to reduce inflammation. antigens labeled with green fluorescent protein to detect patient antibodies
● Therapy with natalizumab: greatly reduces severity of MS and has sensitivity that is similar to RIPA.
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |29
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
Autoimmune
thrombocytopenic Platelets Antiplatelet antibody
purpura
Disseminated
CNS Sensitized T cell
encephalomyelitis
Multisystem
granulomas,
Sarcoidosis Activation of T lymphocytes
pulmonary
manifestations
Autoimmune hemolytic
RBCS Antibody to RBCs
anemia
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |30
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
7. (True or False) Malignant tumors are always encapsulated and rarely spread
UNIT 9C: TUMOR IMMUNOLOGY to adjacent spaces.
I. Pre-test 8. Antigens that can only be found in tumor cells are called: ________________
II. Review of Tumor Biology
a. Characteristics of a Cancer Cell
III. Tumor Antigens REVIEW OF TUMOR BIOLOGY
a. Tumor Specific Antigens (TSA)
b. Tumor Associated Antigens (TAA) ● Tumor immunology is the study of the relationship between the immune
IV. Clinically Relevant Tumor Markers
a. Tumor Markers system and cancer cells
b. Clinical Uses of Tumor Markers ● Salient points:
c. Serum Tumor Markers ○ Tumors arise due to dysregulated growth and impaired apoptotic
V. Laboratory Detection of Tumors mechanisms. These tumors can either be benign or malignant
a. Tumor Morphology
b. Immunohistochemistry ○ Cancer cells have an aberrant DNA. Normally, if cells have an
c. Molecular Methods aberrant DNA or it fails to replicate the correct DNA sequence that
VI. Interaction Between the Immune System and Tumors cell should die. However, some cells escape this mechanism due to
VII. Immunoediting and Tumor Escape the interaction of proto oncogenes and other regulatory
VIII. Immunotherapy
a. Active Immunotherapy mechanisms. It would result in replication with the aberrant DNA.
b. Passive Immunotherapy ○ Malignant tumors are rarely encapsulated and are able to invade
c. Adoptive Immunotherapy the adjacent tissues and other organs
■ Commonly exhibit metastasis
Links
PPT ○ Benign tumors don’t invade
Video ■ Benign tumors are composed of slowly growing cells that
are well differentiated and organized
■ These tumors are surrounded by a capsule, which secures
PRE-TEST them in place and prevents them from circulating to other
parts of the body
1. This tumor marker is used for monitoring hepatocellular carcinoma: ○ Malignant tumors are generally classified into carcinomas,
a. AFP leukemias/lymphomas and sarcomas.
b. hCG ■ Carcinoma- epithelial
c. CA-125 ■ Leukemia- bone marrow
d. CA-19-9 ■ Lymphoma- lymphoid tissue
2. This tumor marker can be used to screen for prostate cancer ■ Sarcoma- mesenchymal
a. AFP ○ Malignant tumors usually arise from a single clone of cells carrying
b. CEA flawed genes/DNA that usually arise from carcinogens
c. PSA ○ The presence of proto-oncogenes and tumor suppressor genes
d. CA-125 highly influence carcinogenesis.
3. (True or False) Tumor markers alone can diagnose the presence of cancer ■ The interplay between proto-oncogenes and tumor
4. (True or False) Tumor cells are polyclonal in nature suppressor genes lead to homeostasis
5. (True or False) Carcinomas are malignant tumors that are mesenchymal in ○ Generally arises from a lack of balance in growth and apoptosis
origin
6. Which of the following is not an oncofetal antigen? CHARACTERISTICS OF A CANCER CELL
a. AFP
b. CEA
● Sustained signaling of proliferation
c. PSA
● Resistance to cell death
d. CA-125
● Ability to induce angiogenesis
● Immortality in terms of cell division
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |31
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
● Invasion and Metastasis ● The only normal cells in which they have been detected are testicular germ
● Ability to avoid suppressors of cell growth cells and placental trophoblastic cells to a lesser extent
● Reprogramming of energy metabolism to support malignant proliferation ● They become TAAs when the transformation process causes them to be
● Ability to evade the immune system expressed on tumors originating from other cell types
● Genomic instability and mutations ○ Example:
● Inflammatory responses that promote tumor growth ■ Melanoma Antigen Gene (MAGE) that are expressed by
melanoma tumors
TUMOR ANTIGENS
DIFFERENTIATION ANTIGENS
● Arsenal in the laboratory
● Used to screen the presence of a tumor or monitor treatment ● Expressed on immature cells of a particular lineage
● Tumor antigens can either be ○ Example:
○ Tumor Specific Antigens (TSA) ■ CD10 antigen which is normally found on pre-B
○ Tumor Associated Antigens (TAA) lymphocytes but not on mature B-lymphocytes
● Identifying the lineage is very important in treating leukemia
TUMOR SPECIFIC ANTIGENS ● Includes oncofetal or embryonic antigens that are normally expressed on
developing cells of the fetus but not on adult cells
● It is thought that genes encoding for these antigens are silent during the
● Unique to a tumor or shared by a limited number of patients with the same development of the embryo and malignant transformation causes them to
type of tumors be re-expressed
● Coded by viral oncogenes or by host proto-oncogenes or tumor suppressor
genes that have undergone mutations
○ Example: ONCOFETAL ANTIGENS
■ Philadelphia chromosome in CML (BCR-ABL gene)
● Some TSAs originate from point mutations in key genes involved in cell ● Carcinoembryonic Antigen (CEA)
proliferation ● Alpha-fetoprotein (AFP)
○ Examples include p53 gene and the gene encoding for Caspase 8 ● Prostatic Specific Antigen (PSA)
● These antigens can be found in the nucleus, cytoplasm, or plasma
membrane of the associated tumor cell OVEREXPRESSED ANTIGENS
● TSAs can also be produced by mutations induced by carcinogenic
chemicals and radiation
● Antigens that arise from genetic mutations that occur during transformation
which results in deregulated expression of these proteins
TUMOR ASSOCIATED ANTIGENS ○ Example:
■ Her2 protein (Human Epithelial Growth Factor Receptor 2)
● These are expressed in tumor cells and normal cells ● Important marker in the treatment of breast
● Tumor cells abnormally express these proteins or carbohydrate antigens in cancer
terms of their concentration, location, or stage of differentiation ● Not limited to breast cancer
● They are increased when tumor is present ● Glycolipid and glycoprotein antigens may be overexpressed in certain
● The normal cell still express this antigen but not that many tumors.
○ Examples:
■ Cancer Antigen 125 (CA-125) and Cancer Antigen 19-9
SHARED TSAs
(CA-19-9)
● Epithelial in origin
● These are expressed in many tumors but not in most normal tissues
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |32
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
● Tumor Markers - Biological substances that are found in increased amounts Proteins Thyroglobulin (TG) Well-differentiated
in the blood, body fluids or tissues of patients with a specific type of cancer. Immunoglobulins (Ig) and Ig papillary or follicular
● These substances can be produced by the tumor itself or by the patient’s light chains (Bence Jones thyroid carcinoma
body in response to the tumor or related benign conditions proteins) Multiple myeloma and
● The concentration of a tumor marker in the serum depends on the degree of lymphoid malignancies
tumor proliferation, the size of the tumor mass, the proteolytic activities of
the tumor, or release of the marker from dying tumor cells Oncofetal antigens Alpha-fetoprotein (AFP) Germ cell carcinomas
● Tumor markers can be proteins, carbohydrates, oncofetal antigens, Carcinoembryonic antigen Hepatocellular carcinoma
hormones, metabolites, receptors, or enzymes (CEA) Colorectal, breast, or lung
cancer
TUMOR MARKERS
Carbohydrate CA 125 Ovarian cancer
antigens CA 15-3 Breast cancer
● Used for screening, diagnosis, or monitoring of treatment CA 19-9 Pancreatic and
● The concentration of a tumor marker in the serum depends on the degree of gastrointestinal cancers
tumor proliferation, tumor mass, the proteolytic activity of the tumor or
release of the marker from the dying tumor cell. Enzymes and Prostate-specific antigens Prostate cancer
● An elevated level of a tumor marker suggests that a significant amount of a isoenzymes (PSA) Bone and liver cancer
particular type of tumor is present. Alkaline phosphatase (ALKP) Neural tissue neoplasms
● Problem: there are elevation even if the patient has no cancer Neuron-specific enolase
IDEAL TUMOR MARKER
Hormones Human chorionic Germ cell carcinoma
gonadotropin (hCG) Trophoblastic tumors
● Should be produced by the tumor itself or by the patient’s body in response Calcitonin Medullary thyroid cancer
to the tumor (unique) Gastrin Pancreatic gastrinoma
● Be secreted into a biological fluid where it can be inexpensively and easily
quantified
● Have a circulating half-life long enough to permit its concentration to rise CLINICAL USES OF TUMOR MARKERS
with increasing tumor load
● Increase to clinically significant levels above the reference level while the
● Four major applications
disease is still treatable
○ Screening
● Have a high sensitivity
○ Diagnosis
● Have a high specificity
○ Prognosis
● Unfortunately very few of the tumor markers in clinical use are ideal because
○ Monitoring
they are not tumor specific
SCREENING
CATEGORIES OF CLINICALLY RELEVANT TUMOR MARKERS
Tumor Marker Class Examples Disease Associations ● Early detection of cancer significantly improves the survival rate
● Tumor markers can be used to screen asymptomatic patients
Cell surface markers Estrogen or progesterone Prognosis for hormone ● Pros:
receptors therapy in breast cancer ○ Screening asymptomatic patients can lead to earlier detection
CD markers on white blood Clonality and lineage of ○ Patients can also be reassured through a negative result
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |33
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
● Cons: ● A significant decrease in the concentration of a tumor marker after surgery,
○ Tends to be expensive. chemotherapy, or other treatment indicates that the therapy has been
○ The possibility of a false negative result can occur. effective in shrinking the tumor
● The effectiveness for screening is dependent on the sensitivity of the test ● An increasing level of the marker after a return to normal indicates that the
and the prevalence of the disease in the population tumor has recurred and that more aggressive treatment may be needed
● Prostatic Specific Antigen
○ Was once highly touted because it is a good marker for prostate
cancer
○ However it is also increased when prostate enlarge (does not
necessarily mean there is cancer)
○ Not used alone
○ Should be accompanied by digital rectal exam
DIAGNOSIS
● A high level of a particular tumor marker usually indicates that the tumor is
aggressive
● Tumor markers can also be used to determine the treatment modality that
best suits the patient
● Can also be indicators for effectivity of a drug
○ Example:
■ Her2 positive breast cancer patients can benefit from
trastuzumab
■ CML- imatinib
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |34
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
NONCANCEROUS
MARKER CANCER(S) USES NORMAL SOURCES CONDITIONS WITH COMMENTS
ELEVATIONS
AFP Nonseminomatous Screening Fetal liver and yolk Pregnancy, non-neoplastic Screening conducted in high - risk populations for
testicular cancer germ cell Diagnosis and sac, adult liver liver disease liver cancer such as those with liver cirrhosis and
Liver staging chronic hepatitis.
Prognosis In germ cell tumors, both AFP and hCG are
Monitoring elevated.
β2-Microglobulin B lymphocyte malignancies Prognosis Part of class I MHC Inflammatory and high cell Higher levels imply poor prognosis in multiple
Monitoring molecule turnover conditions myeloma.
Calcitonin and Familial medullary thyroid Diagnosis Thyroid In hypercalcemia, Can be elevated in other forms of cancer.
Ca++ carcinoma Monitoring increased calcitonin is
expected. Serum Ca++ may
be low when calcitonin is
elevated in medullary
carcinoma.
CD Markers WBC Diagnosis All WBCs WBC increase such as Different CD markers are associated with specific
Monitoring infection WBC malignancies.
CEA Colorectal Prognosis Tissues of Renal failure, Values increased with age and in smokers.
Breast Monitoring endodermal origin non-neoplastic liver and
Lung intestinal disease, age
CA 125 Ovarian adenocarcinoma Screening Ovaries and various Endometriosis, pelvic Increases can occur during menstruation.
Diagnosis other tissues inflammatory disease, Screening is only recommended for women with a
Prognosis uterine fibroids, and family history of ovarian cancer.
Monitoring pregnancy
CA 15-3 Breast cancer Prognosis Mammary tissue Benign breast disease, CA 15-3 is a monoclonal antibody directed against
Can also be increased in Monitoring benign liver disease an epitope of epsilon.
pancreatic, lung, colorectal,
ovarian, and liver cancers
CA 19-9 Pancreatic Diagnosis Sialylated Lewisa Benign hepatobiliary and Can be elevated in some non-pancreatic
Prognosis blood group antigen pancreatic conditions malignancies. Subjects who are Lewisa and b
Monitoring negative persons cannot synthesize CA 19-9
ER/PR Breast adenocarcinoma Prognosis Breast N/A EP/PR+ breast cancers benefit from estrogen or
progesterone reduction therapy.
hCG Nonseminomatous Diagnosis Placenta Pregnancy hCG has a high homology with LH. Malignancies
testicular cancer germ cell Prognosis can produce free α and β chains as well as intact α
trophoblastic (hydatidiform Monitoring and β dimer. Immunoassays that detect only intact
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |35
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
HER2 (neu) or neu Breast Prognosis Growth factor gene in N/A Cancers associated with overexpression of HER2
all cells (neu) or neu have a good response to monoclonal
antibody therapy (trastuzumab).
Monoclonal free Ig Plasma cell, B lymphocytes Diagnosis Normal Igs are Monoclonal gammopathy Bence Jones proteins are free Ig light chains in
light chains Monitoring polyclonal. Few free of undetermined urine detected by urine immunofixation
Prognosis light chains exist. significance (MGUS), electrophoresis.
amyloidosis, nonsecretory
myeloma
Monoclonal Igs Plasma cell, B lymphocytes Diagnosis Normal Igs are Monoclonal gammopathy Monoclonal IgG/IgA → multiple myeloma
Monitoring polyclonal of undetermined Monoclonal IgM → Waldenström’s
Prognosis significance macroglobulinemia
PSA Prostate Screening No tissues other than UTI or prostatitis, benign Levels directly proportional to prostate size. Many
Diagnosis prostate prostatic hypertrophy elevations are benign or not clinically significant.
Monitoring Screening is routinely conducted in men aged 50
Prognosis and older, but is controversial.
Decreased percent of free PSA and PSA velocity
greater than 0.75 ng/mL/year are more strongly
associated with prostate cancer. Collect specimen
before ejaculation, digital rectal examination, or
prostate manipulation.
PTH and CA++ Parathyroid Carcinoma Diagnosis Parathyroid glands In hypocalcemia, increased PTH has a short half-life, so levels are done
Prognosis PTH is expected. Serum intraoperatively to ensure complete parathyroid
Monitoring Ca++ may be high when tumor removal.
PTH is elevated in
parathyroid carcinoma.
TG Thyroid Monitoring Thyroid TG reflects thyroid mass, Assays must simultaneously test for thyroglobulin
injury, and TSH levels. antibodies because these can cause falsely
Thyroid markers (T4, TSH) decreased results. Often tested after TSH
are generally normal in stimulation (or less often, by with-holding thyroid
thyroid cancer. medication) to see if TG production by residual
tumor cells occurs.
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |36
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |37
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
○ Because hCG levels can also increase in men as the result of ● PSA plus histological observations of prostate biopsy can be used to predict
malfunction of the testes the stage of prostate cancer and guide physicians to the best treatment for
● Elevations of hCG can also occur as a result of gonadal suppression caused the patient
by chemotherapy and do not necessarily indicate tumor recurrence ● A rapid rise in PSA or the amount of time it takes to double are indicators of
a more aggressive disease
PROSTATE SPECIFIC ANTIGEN (PSA) ● A persistently high level of PSA after radical prostatectomy indicates
residual disease is present
● Once the tumor is removed the levels of PSA will decrease to undetectable
● It is a 28,000 MW glycoprotein that is produced specifically by epithelial levels
cells in the prostate gland.
● PSA was first discovered in semen, where its function is to regulate the
viscosity of the seminal fluid to facilitate mobility of the sperm cells. LABORATORY DETECTION OF TUMORS
● Widely used marker for prostate cancer
● Largely used for monitoring ● Three types of laboratory methods are routinely used for cancer screening
● Produced specifically by the prostate gland and diagnosis:
● Used in screening for prostate cancer but the practice became controversial ○ Gross and microscopic morphology of tumors
as patients with high levels somewhat underwent unnecessary treatment ■ Most of the time, it identifies the tumor even without the
● Also elevated in benign prostatic hyperplasia (BPH) help of other tumor markers
● Transient increases in PSA also occur after ejaculation, prostate ■ Histopathology
manipulation and digital rectal examination ○ Detection of tumor markers by immunohistochemistry or
● It aids in the diagnosis of prostate cancer automated immunoassays
● Testing now includes PSA-alpha 1-antichymotrypsin complex aside from ○ Molecular diagnostics
testing for free PSA alone which increases the specificity to prostate cancer ■ Confirm the result seen in microscopic morphology
● The proportion of free PSA is higher in benign conditions and the proportion ● The combination of these things makes the diagnosis more accurate
of complexed PSA is higher in malignant conditions
● PSA degrades quickly at temperatures above 4 degrees centigrade. TUMOR MORPHOLOGY
● Testing should be performed within 3 hours of sample collection or to store
the sample at -70 degrees centigrade if a longer interval is needed
● Calculation of the PSA velocity (PSAV) ● Tissue processing of the tumor (histopathology)
○ Calculated as the difference in the PSA concentration divided by ● H&E staining – gold standard in tissue diagnosis
the number of years spanning the interval between sequential tests ● Tumor marker antibodies, special stains and nucleic acid probes can
(reported as ng/mL/year) enhance visible features
○ The rationale for this approach is that the PSA will increase more
rapidly if a growing tumor is present. IMMUNOHISTOCHEMISTRY
○ PSAV greater than 0.75 ng/mL/year was shown to be strongly
associated with the presence of prostate cancer
● Helpful in identifying tumors that looks similar
● PSA density (PSAD)
● Uses labeled antibodies to detect tumor antigens in formalin-fixed tissues or
○ Rationale: An increase in serum PSA is more likely to be caused by
frozen tissue sections.
the occurrence of cancer in a man with a small prostate gland
● Before testing the formalin-fixed tissues should be treated with heat to make
versus a large prostate gland • Calculated as the ratio of the total
antigen epitopes accessible
PSA to the prostate gland volume.
● An unlabeled primary antibody specific for the antigen to be detected is
○ Downside: The need to perform transrectal ultrasonography can be
applied to the tissue section on a slide
costly
● Following an incubation and wash, a labeled secondary antibody directed
● Has an important role in the management of prostate cancer
against the Fc portion of the primary antibody is applied
● The label can be an enzyme such as peroxidase, alkaline phosphatase or
glucose oxidase
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |38
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
● The indirect staining method is used because larger immune complexes are ○ The high-dose hook effect can result in a falsely decreased
formed, providing more sensitive amplification of the signal than is achieved measurement in the area of antigen excess
through direct staining ○ Solution: Dilute the specimen
● Fluorescent dyes can also be used (FITC, rhodamine or Texas red)
● Antigen-antibody reaction would produce color
● This offers a more dynamic range in visualizing the tissue
● Use of positive and negative control tissues is essential for accurate results
○ Negative controls are necessary to ensure that the staining
observed is because of antibody binding and not the background
(i.e., nonspecific reactivity)
○ Positive controls confirm that the antibody reagents are working
properly
● Immunohistochemistry is valuable in certain tumors that have almost similar
histomorphologic features
● Immunohistochemistry : Her2-neu for breast cancer
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |39
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
● Genetic Biomarkers ● In this method, single-stranded DNA or RNA from the tumor is tagged with a
○ Essential for hematologic malignancies (e.g. Philadelphia fluorescent label and incubated with known nucleic acid sequences that
Chromosome) have been spotted onto different areas of a membrane.
○ Detection of gene rearrangements in certain lymphoid neoplasms ● The sample will hybridize to any complementary sequences on the tiny
to determine the lineage spots, allowing for simultaneous testing of the specimen for multiple genes.
○ MSH gene – Familial Colorectal Cancer ○ Aberrant sequences are unique to a particular tumor
○ Estrogen and Progesterone Receptors and Her2 – Breast Cancer. ● Microarrays can also be used to compare the levels of gene expression in
Useful to guide treatment cancer cells with those of normal cells by using two different colors of
○ Methods: FISH, Nucleic Acid Amplification Techniques, Microarray fluorescence to tag nucleic acid from each cell type
and DNA sequencing ● Microarray technology has been developed to test for panels of markers,
○ The aberrant DNA would produce an aberrant mRNA which will rather than individual mutations
produce an aberrant protein
■ With the current technology that we have today, aberrant NEXT GENERATION SEQUENCING
receptors may be characterized
■ Polymerase Chain Reaction: maps the aberrant DNA to
visualize the sequence that is faulty ● Thousands of genes within the tumor can be sequenced simultaneously in
○ Genetic biomarkers can also be used for prospective and post just a few hours to identify genetic variations
diagnostic evaluation of malignancies. ● Can also be used to detect metastases by analyzing DNA from tumor cells
■ Prospective markers can provide valuable information circulating in the peripheral blood
regarding the risk for an asymptomatic person to develop
a particular type of cancer, the growth rate of the cancer, PROTEOMICS
or the development of metastatic disease.
■ Post Diagnostic genetic markers are used to guide
● Analysis of the entire protein complement of a cell population
clinicians in making appropriate treatment decisions for
● Method: Tandem Mass Spectrometry
known cancer patients
● Surface-enhanced laser desorption/ionization mass spectrometry
CYTOGENETICS ● Antibody arrays
● The most common format uses beads that are coated with specific capture
● Karyotyping – To detect aneuploidy, deletions and rearrangements in the antibodies to bind the target proteins and streptavidin- or
chromosomes fluorescent-labeled detection antibodies that can be detected by flow
○ Karyotype analysis has been used for many years to detect the cytometry
chromosomal abnormalities associated with many cancers ● Proteomic methods may allow laboratories to identify unique patterns of
● FISH- can better visualize the rearrangements at a molecular level using proteins and their metabolites that are characteristic of particular types of
probes cancer. This process is called biomarker profiling.
○ Can be used to detect the Philadelphia Chromosome and the
Her2neu gene INTERACTIONS BETWEEN THE IMMUNE SYSTEM AND TUMORS
○ FISH is most often used to detect chromosome rearrangements
and gene amplification.
● One may ask the question “How does our immune system deal with these
tumor cells?”
● The initial theory that tried to explain this is immunosurveillance that runs in
the premise that our immune system is constantly monitoring our body for
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |
40
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
the presence of cancerous and precancerous cells and eliminates them ● Has a limited role in controlling the growth of tumors
before they become clinically evident ● Macrophages activated in vitro by IFNγ have been shown to possess
○ The cells that cannot transform should be killed tumoricidal capabilities
● The immunosurveillance theory was supported by observations that people ● If activated by Interferon Gamma, macrophages exhibit antitumor abilities
who are under immunosuppressive treatment had a higher risk of having ● They appear to kill cancer cells the same way they kill other microorganisms
cancer ● Macrophages produce Tumor Necrosis Factor-Alpha, a cytokine that is
● Also cancer is more prevalent in people 60 years old and above which thought to cause necrosis of tumors by inducing local inflammation and
further supports this theory. thrombosis in the blood vessels within the cancerous mass
● People who are immunosuppressed are prone to have cancer ● Generally, the significance of macrophages in the anti-tumor response
○ HIV+ remains not fully understood
○ Elderly
● Main cells involved: ADAPTIVE IMMUNITY: CYTOTOXIC T-LYMPHOCYTES (CTL)
○ Natural Killer Cells
○ Macrophages
○ Cytotoxic T-lymphocytes (CD8+) ● The primary mechanism of adaptive immunity against tumors is mediated by
○ T-helper lymphocytes (to a certain extent) cytotoxic T lymphocytes (CTLs)
● Antibodies and cytokines play a big role in the immune response to tumor ● Begins with an antigen presenting cell (APC, which is usually a dendritic cell)
● Tumor bearing individuals can also produce antibodies against tumor process tumor antigens in conjunction with class I MHC molecules
antigens. ● The APC presents the tumor-peptide antigen complexes to specific T-cell
○ These antibodies can kill tumor cells by inducing receptors (TCR) on the CTLs and provide co-stimulatory signals that
complement-mediated lysis or ADCC. promote the maturation of the CTLs
○ ADCC occurs when the antibodies coat the tumor cells and bind to ● The mature CTLs use their antigen specific TCRs to bind class I MHC
Fc receptors on the surface of the macrophages, NK cells or associated tumor antigens on the surface of the tumor cell.
neutrophils, stimulating them to release enzymes that can destroy ● Within minutes, their granules migrate toward the plasma membrane and
the tumor targets release cytotoxic proteins within the synapse formed between the CTL and
○ These mechanisms however are clear only in in-vitro studies the target cell
● Dendritic cells are also thought to activate CD4+ Th cells through
presentation of tumor antigens in conjunction with class II MHC molecules
NK-CELLS ○ The activated Th cells may play a role in tumor immunity by
secreting cytokines such as IL-2, which can promote CTL
● Able to kill cells without prior sensitization to tumor antigens development and enhance NK cell activity, and IFNγ, which
● They are activated to kill cells that lack class I MHC molecules which is a activates macrophages and increases class I MHC expression on
feature of transformed cells the tumor cell surface.
● Activating receptors on NK cells bind to tumor antigens or substances
released from the stressed tumor cells, initiating signals that promote CYTOTOXIC T-CELLS
degranulation and the release of perforin and granzymes which ultimately kill
the cell via apoptosis. ● Among the proteins released by the CTL is perforin which creates pores in
● NK cells may also participate in antibody dependent cell cytotoxicity (ADCC) the membrane of the tumor cell, and granzymes which enter the pores and
● in the presence of tumor antibodies cause apoptosis of the tumor target.
● NK cells are thought of to be most effective against malignant cells ● NKT cells – A cell that expresses surface antigens of both CTL and NK cells
circulating in the bloodstream during the early stages of tumor development are able to destroy tumor cells in a mechanism that is similar to CTLs but
● The activity of NK cells can be increased by incubation with IL-2 have a unique type of TCR that recognizes glycolipid antigens instead of
peptide antigens
MACROPHAGES
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |41
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
● Under selective pressure from immunologic forces of attack by cells in the
tumor microenvironment, some of the tumor cells may develop into genetic
T-HELPER CELLS variants that are resistant to immune defenses.
● These cells move past the equilibrium phase and enter the escape phase.
● The cancer cell are thought to enter a state of dynamic equilibrium with the
immune system which keeps the tumor cell under control so that they are
not clinically evident
● During this period, tumor cells may remain dormant or evolve slowly over
time.
● The dynamic interactions between the tumor and the immune system are
thought to shape the phenotype of the tumor and its ultimate outcome,
hence the term immunoediting.
● During this phase mutations can occur in the genetically unstable
transformed cells.
○ When the body is overwhelmed and can’t kill the cell, it would
contain the cell
■ Prone to mutate
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |42
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
PASSIVE IMMUNOTHERAPY
● Expensive
● Involves the administration of soluble components of the immune system to
boost the immune response
● Two approaches to passive immunotherapy in cancer patients involve:
○ Administration of cytokines
○ Monoclonal antibodies
■ MAP
■ Tocilizumab- Covid19 patient
● Cytokines – small proteins that play an important role in regulating the
immune response by serving as chemical messengers.
● Two main applications
○ Used as hematopoietic growth factors
○ Therapeutic Agents
CYTOKINES
IMMUNOTHERAPY
● Hematopoietic growth factor (HGF)/ Colony Stimulating Factors
● In a nutshell, immunotherapy is basically harnessing the ability of the ○ Chemotherapy often suppresses the bone marrow thus decreasing
immune system to kill tumor cells WBC production which are vital for the immune response
● Classified into three (3) types: ○ HGF can be used to prevent this toxicity and to induce the bone
○ Active marrow to produce the much needed WBCs
○ Passive ○ Examples
○ Adoptive ■ GranulocyteColonyStimulatingFactor(G-CSF)
■ Granulocyte-MacrophageColonyStimulatingFactor(GM-CS
F)
■ Erythropoietin
ACTIVE IMMUNOTHERAPY
■ Interleukin-11
● Interferons (IFN)
● Patients are treated in a manner that stimulates them to mount an immune ○ IFN-alpha – most commonly used IFN for cancer therapy
response against their tumors ○ It is thought that IFN-a promotes anti-tumor effects by increasing
● Stimulation of the immune system tumor immunogenicity, enhancement of dendritic cell response,
● Examples: enhances Th1 responses and cell-mediated cytotoxicity, promoting
○ BCG for the treatment of urothelial (bladder) cancer tumor apoptosis and inhibiting angiogenesis
■ Bacille Calmette-Guerin: a vaccine that protects us from ● Interleukins (IL)
extrapulmonary tuberculosis ○ IL-2 is the most extensively studied IL
■ Stimulation of the immune system would help kill urothelial ○ It induces T-cell proliferation and enhancement of CTL and NK-cell
(bladder) cancer function
○ HPV vaccine for cervical cancer ○ Problem: Short half-life and some serious side effects
○ HBV vaccine for prevention of hepatocellular cancer ○ Used only in selected cases (e.g. some cases of melanoma)
■ Hepatitis B destroy hepatocytes
● The intensive repair process tends to create a
mutation that would lead to cancer
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |43
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
MONOCLONAL ANTIBODIES ● Lymphocytes from tumors are harvested and grown in culture and are
administered to the patient. These lymphocytes are thought to have a better
ability to respond to the tumor since they have been exposed to the tumor
● Monoclonal antibodies take a more specific approach to immunotherapy antigens
● These antibodies are derived from a single clone of cells, providing for an ● At present we can now select these lymphocytes, the only ones that can
abundant source of highly specific antibodies directed toward one particular react to the tumor antigen, and selectively culture that clone.
epitope of an antigen ● Downside:
● Generally directed against seven (7) major categories of antigens: ○ HLA typing- body may reject lymphocytes
○ CD antigens ○ Tends to be expensive.
○ Glycoproteins ○ Not all countries have the technology to do such.
○ Glycolipids
○ Carbohydrates
○ Vascular Targets
○ Stromal and Extracellular Antigens
○ Growth Factors
● Some monoclonal antibodies are directed against antigens found on the
surface of the tumor cells.
● These antibodies are believed to destroy the tumor through the same
mechanisms that are used to attack infectious organisms, namely
opsonization, complement-mediated cytotoxicity and ADCC.
● A second group of monoclonal antibodies target surface cell receptors
involved in intracellular pathways that lead to the growth and immortality of
cancer cells
● These antibodies block these known pathways of sustained growth leading
to “immortality”
● A third group of monoclonal antibodies targets antigens involved in
angiogenesis
● These antibodies are directed against vascular endothelial growth factor
(VEGF) ● FIGURE 17–8
● A fourth group boosts the immune response to the tumor by blocking ○ Adoptive immunotherapy with tumor-infiltrating lymphocytes (TILs).
inhibitory pathways that inactivate T-lymphocytes The patient’s tumor is surgically removed and cut into fragments,
● One strategy to increase the effectiveness of monoclonal antibodies involves which are cultured in vitro with IL-2.
linking them to potent cytotoxic drugs that can be taken up by the tumor ○ The cultures are screened for lymphocytes with potent anti-tumor
cells. These products are known as antibody-drug conjugates or activity.
immunotoxins. ○ Positive cultures are expanded further in the presence of IL-2 and
● Limitations to monoclonal antibody treatment: Hypersensitivity reactions are infused into the cancer patient.
○ Before infusion, the patient has been treated with high-dose
ADOPTIVE IMMUNOTHERAPY chemotherapy or radiation to deplete immunosuppressive cells.
● It was discovered that adoptive immunotherapy could be applied to the TABLE 17.8 CANCER IMMUNOTHERAPY USING MONOCLONAL ANTIBODIES
treatment of human cancer TARGET OF MECHANISM OF
● Cells from the immune system are provided to patients. EXAMPLES
THERAPY ACTION
○ Some treatments may combine different types of immunotherapy
● Involves transferring of cells of the immune system to the patient Surface antigens on Opsonization ● Rituximab, a MAb* directed
tumor cells Complement - against the CD20 antigen on B
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |
44
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
mediated cells; used to treat B cell 1. (True or False) T-helper lymphocytes have the biggest role in the immune
cytotoxicity neoplasms. response against tumor cells
ADCC ● Alemtuzumab, a MAb directed 2. These cells can kill tumor cells without prior sensitization of the target cell:
against mature lymphocyte ____________________
antigen, CD52; used to treat 3. Shaping of the phenotype of the tumor cell over time to eventually evade
chronic lymphocytic leukemia and the immune system is called: ___________________
T cell lymphomas 4. Enumerate the phases of immunoediting:
a. Elimination
Cell surface Block signalling ● Panitumumab, a MAb directed b. Equilibrium
receptors pathways involved against epidermal growth factor c. Escape
in cell proliferation receptor (EFGR), used to treat 5. The theory that can explain how the immune system responds to tumor
and survival colorectal cancer. cells: ____________________________
● Trastuzumab, a MAb directed 6. Infusing immune cells to the cancer patient is called:
against HER2, used to treat ____________________________________________
breast and gastroesophageal 7. These antibodies capable of reacting with similar antigens from two or more
tumors with overexpressed HER2. unrelated species:___________________________
8. (True or False). Majority of tumor markers have high sensitivity and
Antigens involved in Inhibit formation of ● Bevacizumab, a MAb directed
specificity
angiogenesis blood vessels against vascular endothelial
necessary for growth factor (VEGF); for
delivery of oxygen treatment of glioblastoma, colon,
and nutrients to the lung, and renal cancers.
tumor
POST-TEST
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |45
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
TRANSPLANTATION
● The HLA disparity between donor and recipient that occurs with allografts
and xenografts will result in a vigorous cellular and humoral immune
response to foreign MHC antigens- this response is the primary stimulus
of graft rejection.
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |
46
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |47
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
similarities in the structure of the allo-HLA protein itself or to structural
similarities of allo-HLA protein + peptide.
● Virus-specific T cells may be an important source of alloreactive cells.
● Characterized by a high frequency of responding T cells compared with the
responder frequency in a typical T-cell response to a foreign antigen.
● Mixed Lymphocyte Reaction (MLR)
○ In vitro correlate of direct allorecognition
○ Lymphocytes from an individual needing a transplant are
incubated with lymphocytes from a potential donor that
have been inactivated so they cannot proliferate.
● A high level of radioactivity indicates that the recipient’s T cells have divided
in response to different HLA-D antigens on a potential donor’s cells and that
such a donor would be more likely to stimulate graft rejection.
RECOGNITION OF ALLOANTIGENS
DIRECT RECOGNITION
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |
48
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
INDIRECT RECOGNITION
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |
49
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
● Occurs within minutes to hours after the vascular supply to the transplanted
organ is established.
● Mediated by a preformed antibody that reacts with donor vascular
endothelium.
● ABO, HLA and certain endothelial antigens may elicit hyperacute rejection.
○ Antibodies may be present as a result of blood
transfusion, prior transplantation, or exposure of a
pregnant woman to fetal antigens of paternal origin.
● Binding of preformed antibodies to the alloantigens activates the
complement cascade and clotting mechanisms and leads to thrombus
formation.
○ Results to ischemia and necrosis of the transplanted
tissue.
● Seldom encountered in clinical transplantation
● Absence of donor HLA-specific antibodies is confirmed before transplant by
performing crossmatch test.
HYPERACUTE REJECTION
● Accelerated Rejection
○ Antibody-mediated rejection may take place over several
1. Preformed Ab days.
● Ab that are already presented in the circulation. ○ Occur in individuals who possess low levels of
● Transfusion of incompatible blood types. donor-specific antibody in the pretransplant period.
2. Complement activation ○ Involves intravascular thrombosis and necrosis of donor
● Interaction between the preformed Ab and antigen will activate the tissue.
complement which will result in lysis.
3. Neutrophil margination
ACUTE REJECTION
4. Inflammation
5. Thrombosis formation
● Follows the usual recognition of foreign substance and then produce
antibodies.
● It takes weeks to months because there will be processing unlike in
hyperacute where Ab is already present.
● The larger the tissue, the faster the rejection.
○ Immunosuppressants are usually given.
● Mediated by a cellular alloresponse (ACR) or by donor-specific antibody
(aka antibody-mediated response; AMR)
1. T-cell, macrophage and Ab mediated
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |50
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
2. Myocyte and endothelial damage
3. Inflammation
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |51
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
ORGAN TRANSPLANTATION
● Heart
● Lungs
● Kidney
● Liver
● Skin
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |52
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
● Theory before is that these sites are protected by barriers that protect them IMMUNOSUPPRESSIVE AGENTS
from the immune system but later on found out that these areas have certain
mechanisms that prevent them from induction of immune response.
● Example: ● May be used in induction and maintenance of immune suppression and
○ Eyes treatment of rejection.
■ Lacks lymphatic vessels ● Combinations of different agents are frequently used to prevent graft
■ Low levels of MHC molecules infection.
■ High expression of CD59 and DAF - serves as inhibitors of ● Immunosuppressive state (and graft survival) induced by these agents
MAC comes at a price of increased susceptibility to infection, malignancies, and
○ Placenta other associated toxic side effects.
○ Testicles
○ Central nervous system 3 MAIN IMMUNOSUPPRESSIVE DRUGS
1. Cyclosporins
● Act by inhibiting T-cell activation, thus preventing T-cells from
attacking the transplanted organ.
2. Azathioprine
● Mitotic inhibitor
3. Corticosteroids
● Such as prednisolone suppress the inflammation associated with
transplant rejection.
● Potent anti-inflammatory and immunosuppressive agents used for
immunosuppressive maintenance.
● At higher doses, they are used to treat AR episodes.
● Act by blocking production and secretion of cytokines,
inflammatory mediators, chemoattractants and adhesion
molecules.
● These activities decrease macrophage function and alter
leukocyte-trafficking patterns.
● Long-term use is associated with hypertension and diabetes
mellitus.
● Some immunosuppressants are also used to treat a variety of autoimmune
diseases:
○ Azathioprine in treatment of rheumatoid arthritis
○ Cyclosporin is used in heart, liver, kidney, pancreas, bone marrow
and heart/lung transplantation
IMMUNOSUPPRESSION ○ Glatiramer acetate is used in treatment of relapsing-remitting
multiple sclerosis.
● Immunosuppression can be brought about by 3 different ways: ○ Mycophenolate is used to prevent the kidney problems associated
○ Surgical ablation with lupus erythematosus.
○ Lymphoid irradiation ○ Sirolimus used for the treatment of psoriasis.
○ Immunosuppressive drugs
CLASSES OF IMMUNOSUPPRESSIVE AGENTS
● Antimetabolites
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |53
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
○ interfere with the maturation of lymphocytes and kill proliferating
cells.
○ Azathioprine
■ First agent employed
● Calcineurin inhibitors
○ Cyclosporine and FK-506 (tacrolimus) are compounds that block
signal transduction in T lymphocytes resulting in impaired synthesis
of cytokines (IL-2, IL-3, IL-4 and interferon gamma).
○ Inhibition of cytokine synthesis blocks the growth and
differentiation of T cells, impairing anti graft response.
● Monoclonal antibodies
○ Bind to cell surface molecules on lymphocytes are used at the time
of organ transplant and to treat severe rejection episodes after
transplantation.
○ Basiliximab and daclizumab
■ Both bind the CD25 (IL-2 receptor) and thus interfere with
the IL-2 mediated T-cell activation.
● Polyclonal antibodies
○ Thymoglobulin
■ An antithymocyte antibody prepared in rabbits
○ ATGAM
■ Polyclonal antiserum prepared from the immunization of
horses.
○ Both are potent immunosuppressive agents that deplete
lymphocytes from the circulation.
POST TEST
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |54
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
I. Introduction
II. Deficiencies Of B-cell System ● At birth, IgG is about the same as adult level due to the maternal transfer but
III. Deficiencies Of Cellular Immunity slow decrease due to catabolism over 2-3 months
IV. Combined Deficiencies Of Cellular And Humoral Immunity ● For the first few months of age, own antibodies gradually rise in response to
V. Defects Of Neutrophil Function environmental stimuli
VI. Complement Deficiencies
VII. Laboratory Evaluation Of Immune Dysfunction ● Late increase in the levels of Immunoglobulins
○ IgM and IgA
Links ■ Not affected
PPT ■ IgM reaches the normal adult level at 1 year of age
Video
○ IgG
■ 2SD (standard deviation) levels below normal
■ Lasts 9-15 months of age
INTRODUCTION
● Signs and symptoms:
○ Pyogenic sinopulmonary and skin infection
● Components of the immune system play unique but overlapping roles in the
● This condition does not appear to be X-linked, although it is more common
host-defense process
in males.
● Interact extensively through many regulatory and effector loops
● Immunodeficiency Disorders
○ Decreases ability to defend against infectious organisms BRUTON’S AGAMMAGLOBULINEMIA
○ Increases susceptibility to develop certain types of malignancies
○ Clinical symptoms range from very mild/subclinical to severe ● Agammaglobulinemia - genetic defect in B cell maturation or mutation
○ Recurrent infections or failure to thrive may also happen leading to the defective interactions between B and T cells
○ Can be inherited or acquired secondary to other conditions such as ● X-linked
certain infections, malignancies, autoimmune disorders, and ● Deficiency/lack of all classes of immunoglobulins
immunosuppressive therapy ○ Lack circulating mature CD19+ B cells
○ Ineffective immune response ■ Pre-B cells in bone marrow but no peripheral cells in
○ Due to a missing or deficient component/s of the immune system lymphoid tissues
○ Secondary immunodeficiencies ■ The differentiation stops at pre-B cell stage due to the
■ Acquired immunodeficiency syndrome (AIDS), which is deficiency in Bruton’s tyrosine kinase
caused by the human immunodeficiency virus (HIV) ● Bruton’s tyrosine kinase (BTK) - enzyme responsible for the VH gene
○ Primary immunodeficiency (PIDs) rearrangement
■ Inherited dysfunctions of the immune system ● Treatment: IM/IV Ig
● Defect in one arm of the immune system may affect other aspects of
immune function
SELECTIVE IGA DEFICIENCY
● Deficiency of one component of the system is accompanied by hyperactivity
of other components
● Most common congenital immunodeficiency
● Most patients with IgA deficiency are asymptomatic
DEFICIENCIES OF B-CELL SYSTEM (AGAMMAGLOBULINEMIA)
● Impaired differentiation of lymphocytes to become IgA-producing peripheral
cells
1. Transient Hypogammaglobulinemia of Infancy
2. Bruton’s Agammaglobulinemia ○ 30-40% of patients develop anti-IgA antibodies
3. Selective IgA Deficiency ● Signs and symptoms:
4. Common Variable Immunodeficiency ○ Infections in respiratory and gastrointestinal tract
5. Isolated IgG Subclass Deficiency ○ Increased tendency to develop autoimmune diseases such as SLE,
Rheumatoid arthritis, and other immunodeficiency diseases
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |55
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
○ Allergic disorders and malignancy are also most common X-linked All antibody Pre-B cell Infancy
agammaglobulinemia isotypes reduced differentiation
COMMON VARIABLE IMMUNODEFICIENCY (CVI)
Reduced B cell; excess T Usually 20-30
Common variable antibody; many suppression years of age
● A heterogeneous group of disorders immunodeficiency different
● Most common primary immune deficiency with a severe clinical syndrome combinations
● The disorder can be congenital or acquired, or familial or sporadic, and it
occurs with equal frequency in men and women Reduced IgG1, Defect of isotype Variable with the
Isolated IgG subclass
● Deficiency in both IgA and IgG IgG2, IgG3, or differentiation class and degree
deficiency
○ Selective IgG deficiency may occur IgG4 of deficiency
● Signs and symptoms:
Immunodeficiency Reduced IgG, B-cell switching Infancy
○ Recurrent bacterial infections (sinusitis, pneumonia)
with IgA, IgE, with
● 3 cellular defects:
hyperimmunoglobulin elevated IgM
○ T cells or their products appear to suppress differentiation of B
M
cells
○ T cells fail to help in B cell terminal differentiation
○ Primary defect in B cell line DEFICIENCIES OF THE CELLULAR IMMUNITY
● CVI is diagnosed by demonstrating a low serum IgG level in patients with
recurrent bacterial infections. ● T cells: cell-mediated immunity
● Treatment: IM/IV Ig 1. DiGeorge Anomaly
2. Purine Nucleoside Phosphorylase Deficiency
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |56
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
● Produces a moderate to severe defect in cell-mediated immunity with
normal or only mild impaired humoral immunity JAK3 DEFICIENCY (AUTOSOMAL RECESSIVE)
● Signs and symptoms:
○ Recurrent or chronic pulmonary infections
○ Oral or cutaneous candidiasis ● T cell- B cell+ NK cell-
○ Diarrhea ● Affect both males and females.
○ Skin infections ● Deficiency in intracellular kinase JAK3 (required for processing interleukin
○ UTI binding signal from cell membrane to the nucleus)
○ Failure to thrive ○ No signal transmission from IL2 and IL4 in lymphocytes
● Symptoms are similar to the X-linked SCID
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |57
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
○ Located on the X chromosome, region p11. RAG1/2; JAK3;
common gamma
chain receptor
● Rare autosomal recessive syndrome characterized by: AT Reduced IgG2, IgA, DNA instability Infancy
○ Cerebral ataxia - involuntary muscle movement IgE, and T
○ Telangiectasia - capillary swelling resulting in red patches on skin lymphocytes
especially in the earlobes and conjunctiva
Reticular All leukocytes Stem cell defect Neonatal
● Blunt Ab response to Ags, especially polysaccharides
dysgenesis
○ Low or absent IgG2, IgA, IgE
● Decreased circulating T cells
○ Rearrangement of TCR and IgG genes does not occur DEFECTS OF NEUTROPHIL FUNCTION
○ Usually occurs in early adults from either pulmonary disease or
malignancy 1. Chronic Granulomatous Disease
● Treatment: 2. Neutrophil G6PD deficiency and myeloperoxidase deficiency
○ Bone marrow transplant/cord blood stem cells 3. Leukocyte Adhesion Deficiency
● AT gene
○ Located on chromosome 11, region q22. CHRONIC GRANULOMATOUS DISEASE (CGD)
COMBINED DEFICIENCIES OF CELLULAR AND HUMORAL IMMUNITY ● A group of disorders involving inheritance of either an X-linked or autosomal
recessive gene that affects the neutrophil microbicidal function
● Most common and best characterized of the neutrophil abnormalities
CHARACTERISTICS OF SELECTED DEFECTS OF THE T-CELL SYSTEM AND ● Inability of neutrophil to produce reactive forms of oxygen necessary for
COMBINED DEFECTS bacteria killing
CONDITION DEFICIENCY LEVEL OF PRESENTATION ○ No oxidative (respiratory) burst
DEFECT ○ Diagnosed by measuring the ability of the patient’s neutrophil to
reduce the nitroblue tetrazolium dye
DiGeorge anomaly T cells; some Embryological Neonatal, with ● Symptoms:
secondary effects development of the hypocalcemia or ○ Recurrent suppurative infections
on antibody thymus cardiac defects if ○ Pneumonia
production severe; incomplete ○ Osteomyelitis
forms may be ○ Draining adenopathy
present later with ○ Liver abscesses
infection ○ Dermatitis
○ Hypergammaglobulinemia
PNP deficiency T cells; some PNP, purine Infancy
● Treatment:
secondary effects metabolism
○ Bone marrow transplant/PBSC
on antibody
○ Granulocyte transfusion
production
○ Cytokine administration
SCID Both T and B cells ADA, purine Infancy ○ Antibiotics
metabolism; HLA
expression; NEUTROPHIL GLUCOSE-6-PHOSPHATE DEHYDROGENASE DEFICIENCY
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |58
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |59
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
CONFIRMATORY TESTS
● Serum proteins are electrophoresed
● Specific antibodies allow to bind on the separated proteins
FLOW CYTOMETRY ● Interpret band formation
○ Diffuse: polyclonal immunoglobulin
● Enumeration of classes and subclasses of lymphocytes through CD markers ○ Narrow, intense: monoclonal
○ CD19: B cells ● Lack of bands indicates immunodeficiency of one or more immunoglobulin
○ CD3: T cells classes.
■ CD3/CD4: Th cells
■ CD3/CD8: Tc cells BONE MARROW BIOPSY
○ CD16 or CD56: NK cells
● Eg., low CD19 -> suggests Bruton’s agammaglobulinemia ● Monoclonal gammopathy
● Objective and quite reliable in detecting defects that result in a decrease in ● Immunodeficiency state
one or more types of lymphocytes.
T-CELL FUNCTION (CLASSICAL) POST TEST
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |
60
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
■ Chemokines attract the cell at the site of infections and
UNIT 10: IMMUNOMODULATION the regulatory cytokines (eg. interleukin) helps control the
I. Immunomodulation inflammation
a. Immunomodulators ● Proinflammatory and antiviral cytokines induced by immunomodulators
b. Non-specific immunomodulation by IV immunoglobulin such as OM-85 as part of the innate response include IL-1β, IL-6, and tumor
c. Monoclonal antibodies for specific immunomodulation necrosis factor α, and IL-12, interferon (IFN)-α, IFN- β, IFN-γ, respectively.
Links ● Bacterial lysate immunomodulators also induce cytokines related to the
PPT adaptive immune system including the B cell-activating cytokines IL-10, B
Video cell activating factor (BAFF), and IL-6.
○ The activation of the adaptive T helper cell and B cell classes of
immunomodulators has been demonstrated as the immunoglobulin
PRE-TEST response cascade of rapid IgM production, followed by IgA and
IgG.
1. Drugs that either suppress or stimulate the immune system ○ Alongside IgA, which is central to the mucosal immune response,
2. Substances that stimulate the immune system by inducing activation and IgG has the advantage of being more widely distributed and
increasing the activity of any of its components. providing a longer humoral memory response.
3. Suppress the immune system ● The immunoregulatory effects of bacterial lysate immunomodulators involve
maturation of both plasmacytoid and myeloid dendritic cells, indicated by
the presence of T cell regulatory proteins CD80 and CD86.
IMMUNOMODULATION
IMMUNOMODULATORS
● Change in the body's immune system, caused by agents that activate or
suppress its function.
● Immunomodulation by bacterial lysates involves both induction of immune ● Drugs that either suppress or stimulate the immune system
system effector cells and activation of immunoregulatory cell classes.
○ This effect mirrors commensal microorganisms which both IMMUNOSTIMULANTS
stimulate immune system maturation and reduce allergic
sensitization.
● Are substances that stimulate the immune system by inducing activation
○ Bacterial lysates immunomodulators induce immune effector cells
and increasing the activity of any of its components.
hence reducing the infection and activates the immunoregulatory
● They are used in disorders which include immunodeficiency diseases,
cells thus reducing inflammation.
malignancy, viral, fungal and certain autoimmune disorders.
● Evidence from in vitro, in vivo, and human trials indicates that the
immunomodulatory effects of bacterial lysates induce effector cells in both
the innate and adaptive immune system and their respective regulatory IMMUNOSUPPRESSION/IMMUNOSUPPRESSANTS
dendritic cell and regulatory T and B cell populations.
● Much of the immunomodulatory response is dependent on the previously ● Cytotoxic agents
mentioned TLR, expressed on epithelial cells, dendritic cells, macrophages, ● Inhibition of lymphocyte signalling
monocytes, and T and B lymphocytes. ● Cytokine inhibitors
○ TLR (toll-like receptor) ● Depletion of specific immune cells
■ Class of proteins that plays a role in the innate immune ● Blockade of cell adhesion
system. ● Inhibition of complement activation
■ The type of TLR that is activated determines the
downstream activity, thus the production of cytokines is
based on immunoinflammatory responses against the NON-SPECIFIC IMMUNOMODULATION
pathogen.
● Facilitated by intravenous immunoglobulin.
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |61
MT6326 | Immunology and Serology Lecture
1st Shifting (A.Y. 2020 - 2021)
Mr. Benjie M. Clemente, RMT, MLS (ASCPi)cm, MPH
● Immunoglobulin replacement is essential for patients with primary antibody
deficiencies and of proven value in several forms of secondary
hypogammaglobulinemia.
● Beneficial effect of IVIG ,which has been established by control trials against
the placebo or conventional treatment in several disorders.
○ Benefit has been claimed in open trials which provides inclusive
evidence for general use.
○ Mechanism of IVIG as the treatment of choice against Kawasaki
disease in children is unknown.
■ Neutralization of the unknown infectivity plays a role in the
treatment of Kawasaki disease.
● IVIG raised the platelet count in two hypogammaglobulinaemic children with
idiopathic thrombocytopenia inspired a new approach to the therapy of
autoimmune disease.
MONOCLONAL ANTIBODIES
References: PPT, Clinical Immunology and Serology: A Laboratory Perspective , Lecturer 3OMT | CASTILLO, C. | CASTILLO, E. | HERNAEZ | KASILAG | LAGMAN | ROSALES | TORRES |62