UNIVERSIDAD COLEGIO MAYOR DE CUNDINAMARCA

INMUNOLOGIA GENERAL

ARTICULOS EN INGLES
TEMA: CMH EN PORCINOS

PROFESORA: JEANNETTE NAVARRETE OSPINA

INTEGRANTES:

BAYRON STEBAN DAZA MARTINEZ

JEIMY VALENTINA DUARTE ROMERO
JAVIER STEVEN LONDOÑO P

2016
1. Article

Porcine major histocompatibility complex (MHC) class I molecules

and analysis of their peptide-binding specificities
Lasse Eggers Pedersen,Mikkel Harndahl, Michael Rasmussen, Kasper
Lamberth, William T. Golde, Ole Lund, Morten Nielsen, and Soren Buus.

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3214623/

2011

In all vertebrate animals, CD8+ cytotoxic T lymphocytes (CTLs) are controlled by

major histocompatibility complex class I (MHC-I) molecules. These are highly
polymorphic peptide receptors selecting and presenting endogenously derived
epitopes to circulating CTLs. The polymorphism of the MHC effectively
individualizes the immune response of each member of the species. We have
recently developed efficient methods to generate recombinant human MHC-I (also
known as human leukocyte antigen class I, HLA-I) molecules, accompanying
peptide-binding assays and predictors, and HLA tetramers for specific CTL staining
and manipulation. This has enabled a complete mapping of all HLA-I specificities
(“the Human MHC Project”). Here, we demonstrate that these approaches can be
applied to other species. We systematically transferred domains of the frequently
expressed swine MHC-I molecule, SLA-1*0401, onto a HLA-I molecule (HLA-
A*11:01), thereby generating recombinant human/swine chimeric MHC-I molecules
as well as the intact SLA-1*0401 molecule. Biochemical peptide-binding assays
and positional scanning combinatorial peptide libraries were used to analyze the
peptide-binding motifs of these molecules. A pan-specific predictor of peptide–
MHC-I binding, NetMHCpan, which was originally developed to cover the binding
specificities of all known HLA-I molecules, was successfully used to predict the
specificities of the SLA-1*0401 molecule as well as the porcine/human chimeric
MHC-I molecules. These data indicate that it is possible to extend the biochemical
and bioinformatics tools of the Human MHC Project to other vertebrate species.
2. Article

Predicting MHC haplotypes from high-density SNP genotypes in

pigs

J.R. Dunkelberger* , C.S. Ho† , A.S. Hess* , N.V.L. Serão* , J.K. Lunney‡ , and
J.C.M. Dekkers* * Department of Animal Science, Iowa State University, Ames, IA;
† Gift of Life Michigan, Ann Arbor, MI; ‡ USDA, ARS, BARC, APDL, Beltsville, MD

https://asas.org/docs/default-source/wcgalp-
posters/532_paper_9222_manuscript_496_0.pdf?sfvrsn=2

2011

ABSTRACT

The major histocompatibility complex (MHC) region in swine contains genes

associated with disease resistance, many of which have haplotype-specific
expression. The objective of this study was to determine whether SNP genotypes
from a high-density SNP chip can be used to predict haplotypes within the MHC
region of the swine genome. In total, 920 pigs were genotyped using the Illumina
SNP60 BeadChip. SNPs located within the Swine Leukocyte Antigen (SLA) I and
SLA II regions of chromosome 7 were used to define SNP haplotypes for MHC
classes I and II, respectively, using BEAGLE phasing software. Each SNP
haplotype corresponded uniquely to a MHC haplotype defined by a PCR-based
method. In conclusion, highdensity SNP genotypes from the Illumina Porcine
SNP60 BeadChip can be used to predict MHC haplotypes with fairly high accuracy,
as an alternative to PCR-based approaches.

3. Article

Molecular genetics of the swine major histocompatibility complex,

the SLA complex
Joan K. Lunney, Chak-Sum Ho, Michal Wysocki, Douglas M.Smith
http://www.umass.edu/vetimm/publications/Lunney_etal_2009.pdf

2010

ABSTRACT
The swine major histocompatibility complex (MHC) or swine leukocyte antigen
(SLA) complex is one of the most gene-dense regions in the swine genome. It
consists of three major gene clusters, the SLA class I, class III and class II regions,
that span 1.1, 0.7 and 0.5 Mb, respectively, making the swine MHC the smallest
among mammalian MHC so far examined and the only one known to span the
centromere. This review summarizes recent updates to the Immuno Polymorphism
Database-MHC (IPD-MHC) website (http://www.ebi.ac.uk/ipd/mhc/sla/) which
serves as the repository for maintaining a list of all SLA recognized genes and their
allelic sequences. It reviews the expression of SLA proteins on cell subsets and
their role in antigen presentation and regulating immune responses. It concludes
by discussing the role of SLA genes in swine models of transplantation,
xenotransplantation, cancer and allergy and in swine production traits and
responses to infectious disease and vaccines.

4. Article

Human monoclonal HLA antibodies reveal interspecies

crossreactive swine MHC class I epitopes relevant for
xenotransplantation
Arend Mulder, Marrie J. Kardol, J. Scott Arn, Chantal Eijsinka, Marry E.I. Frankea,
Geziena M.T. Schreuder, Geert W. Haasnoot, Ilias I.N. Doxiadis, David H. Sachs,
Douglas M. Smith, Frans H.J. Claas

http://www.epitopes.net/pdfs/other/q50Mulder.pdf

2010

ABSTRACT

Crossreactivity of anti-HLA antibodies with SLA alleles may limit the use of pig
xenografts in some highly sensitized patients. An understanding of the molecular
basis for this crossreactivity may allow better selection of xenograft donors. We
have tested 68 human monoclonal HLA class I antibodies (mAbs) for reactivity with
pig lymphocytes from SLA defined pigs and found nine to be crossreactive. Eight of
nine were broadly HLA reactive IgM-mAbs. The putative HLA epitopes for seven
mAbs. were conserved in the aminoacid sequence of the SLA alleles studied. The
lack of reactivity of a large number of mAbs largely correlated with the absence of
the putative epitopes in the SLA alleles studied. We conclude that most patients
with anti-HLA class I antibodies should be able to find pig donors lacking SLA
antigens that cross react with their antibodies and that many of the crossreacting
epitopes can be defined by analysis of shared epitopes in the aminoacid sequence
of human and pig MHC antigens.

5. Article
 Relationships between the polymorphism of
myosin heavy chains and selected meat quality
traits of pigs with different susceptibility to stress

BOŻENA GRZEŚ, EDWARD POSPIECH,MARIA KOĆWIN-PODSIADŁA,

ANDRZEJ ŁYCZYŃSKI, ELŻBIETA KRZĘCIO, BEATA MIKOŁAJCZAK and EWA
IWAŃSKA.
http://www.archanimbreed.com/pdf/2010/at10p065.pdf

2010

ABSTRACT

The aim of the investigations was to analyse the share of myosin heavy chains
(MHC) isoforms (type I, IIa, IIb, and IIx) in the longissimus thoracis et lumborum
muscle derived from pigs of different RYR1 genotypes (TT – homozygous
negative, CT – heterozygous, CC – homozygous positive). The composition of the
MHC isoforms in the muscle tissue of the examined animals was referred to
selected meat quality traits. It was revealed that the animals with the CT and TT
genotypes were characterized by a significantly (P≤0.05) lower share of the type I
and higher share of the type IIb MHC isoform in comparison to homozygotes CC.
Inferior tenderness and water holding capacity of meat obtained from pigs
susceptible to stress (TT) at 144 h after slaughter could have been associated,
among others things, with the increased share of MHC isoform type IIb. The
composition of MHC isoforms might be a useful indicator in breeding work in the
selection of animals carrying the gene of susceptibility to stress.

6. Article

Transcription variants of SLA-7, a swine non classical MHC class I

gene
Rui Hu,Gaëtan Lemonnier, Emmanuelle Bourneuf, Silvia Vincent-
Naulleau, and Claire Rogel-Gaillard.

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3108204/

2011

ABSTRACT
In pig, very little information is available on the non classical class I (Ib) genes of
the Major Histocompatibility Complex (MHC) i.e. SLA-6, -7 and -8. Our aim was to
focus on the transcription pattern of the SLA-7 gene. RT-PCR experiments were
carried out with SLA-7 specific primers targeting either the full coding sequence
(CDS) from exon 1 to the 3 prime untranslated region (3UTR) or a partial CDS from
exon 4 to the 3UTR. We show that the SLA-7 gene expresses a full length
transcript not yet identified that refines annotation of the gene with eight exons
instead of seven as initially described from the existing RefSeq RNA. These two
RNAs encode molecules that differ in cytoplasmic tail length. In this study,
another SLA-7 transcript variant was characterized, which encodes a protein with a
shorter alpha 3 domain, as a consequence of a splicing site within exon 4.
Surprisingly, a cryptic non canonical GA-AG splicing site is used to generate this
transcript variant. An additional SLA-7 variant was also identified in the 3UTR with
a splicing site occurring 31 nucleotides downstream to the stop codon. In
conclusion, the pig SLA-7MHC class Ib gene presents a complex transcription
pattern with two transcripts encoding various molecules and transcripts that do not
alter the CDS and may be subject to post-transcriptional regulation.

7. Article

Intracellular localization and association of MHC class I with

porcine invariant chain

F.Z. Xu, S.G. Wu and W.Y. Yu.

http://www.funpecrp.com.br/gmr/year2013/vol12-1/pdf/gmr2180.pdf

2013

ABSTRACT

The objective was to investigate the intracelular localization and association of pig
major histocompatibility complex (MHC) class I subunits with invariant chain (Ii).
Pig MHC class I subunit cDNAs were cloned by RT-PCR and eukaryotic
expression plasmids of α and β2m were constructed with fusions to red or
enhanced green fluorescent protein (pDsRed2-N1-α, pEGFP-N1-α, pDsRed2-N1-
β2m, and pEGFP-N1-β2m). A pig Ii mutant with a deleted CLIP región (DCLIP-Ii)
was constructed by overlap extension PCR. Wild-type Ii and mutant Ii were cloned
into pEGFP-C1 (pEGFP-C1-Ii, pEGFP-C1- DCLIP-Ii). The recombinant plasmids of
MHC I subunits and pEGFPC1-Ii (pEGFP-C1-DCLIP-Ii) were transiently
cotransfected into COS-7 cells with Lipofectamine 2000. Immunofluorescence
microscopy was performed to detect expression and intracellular localization of Ii
and MHC I subunits, and immunoprecipitation was used to analyze their
association. Our results indicated that pig Ii associates with integrated MHC I
subunits to form oligomers, but cannot associate with single MHC I subunits.
Furthermore, deletion of the Ii CLIP sequence blocks association with integrated
MHC I subunits. Thus, pig Ii cannot associate with a single MHC I molecule, the α
or β2m chain, but Ii and the integrated MHC I molecule can form complexes that
colocalize in the endomembranes of COS-7 cells. The Ii of CLIP plays a key role in
assembly of Ii and MHC I.

8. Article

Sequences polymorphism and variation of major

histocompatibility complex DRB exon 2 of black Dahe
pig

Lizhou Tang, Long Yu, Jiangang Chen, Junjie Wang, Mei Ma, Weidong Lu and
Tongzuo Zhang.

http://www.academicjournals.org/AJB

2012
ABSTRACT

DNA sequences of swine leukocyte antigens (SLA) DRB exon 2 from five
populations were used to investigate genetic polymorphism of black Dahe pig in
Yunnan province of China. Our results showed that the SLA-DRB exon 2 of black
Dahe pig obtained high variable rate of 23.50%, which was higher than the values
analyzed for cattles or other livestocks. The average G + C content at the third
synonymous variable coding positions (GC3) was closed to 0.78. The amino acids
composition rate and majority of relative synonymous codon usage were near the
ranges of 0.00 to 11.27 and 1.00 to 2.00, respectively. These significant
polymorphism, high GC3 content and differentiation of preferred codón usage
could be resulted from over dominance selection, base compositional bias and the
diversity of gene expression level. Aforementioned results adequately testified the
SLA-DRB exon 2 may be an important genetic marker for studies in genetic
diversity, population genetics, breeding reproduction investigation and exploring
new functional genes of black Dahe pig.

9. Article

Establishing the pig as a large animal model for vaccine

development against human cancer
Nana H. Overgaard, Thomas M. Frøsig, Simon Welner , Michael Rasmussen,
Mette Ilsøe, Maria R. Sørensen, Mads H. Andersen, Søren Buus and Gregers
Jungersen

http://journal.frontiersin.org/article/10.3389/fgene.2015.00286/abstract

2015

ABSTRACT
Immunotherapy has increased overall survival of metastatic cancer patients, and
cancer antigens are promising vaccine targets. To fulfill the promise, appropriate
tailoring of the vaccine formulations to mount in vivo cytotoxic T cell (CTL)
responses toward co-delivered cancer antigens is essential. Previous development
of therapeutic cancer vaccines has largely been based on studies in mice, and the
majority of these candidate vaccines failed to induce therapeutic responses in the
subsequent human clinical trials. Given that antigen dose and vaccine volume in
pigs are translatable to humans and the porcine immunome is closer related to the
human counterpart, we here introduce pigs as a supplementary large animal model
for human cancer vaccine development. IDO and RhoC, both important in human
cancer development and progression, were used as vaccine targets and 12 pigs
were immunized with overlapping 20mer peptides spanning the entire porcine IDO
and RhoC sequences formulated in CTL-inducing adjuvants: CAF09, CASAC,
Montanide ISA 51 VG, or PBS. Taking advantage of recombinant swine MHC class
I molecules (SLAs), the peptide-SLA complex stability was measured for 198 IDO-
or RhoC-derived 9-11mer peptides predicted to bind to SLA-1∗04:01, −1 ∗07:02,
−2 ∗04:01, −2 ∗05:02, and/or −3 ∗04:01. This identified 89 stable (t½ ≥ 0.5 h)
peptide-SLA complexes. By IFN-γ release in PBMC cultures we monitored the
vaccine-induced peptide-specific CTL responses, and found responses to both
IDO- and RhoC-derived peptides across all groups with no adjuvant being superior.
These findings support the further use of pigs as a large animal model for vaccine
development against human cancer

10. Article

Pig Models of Neurodegenerative Disorders: Utilization in Cell

Replacement-Based Preclinical Safety and Efficacy Studies

Dasa Dolezalova, Marian Hruska-Plochan, Carsten R. Bjarkam, Jens Christian H.

Srensen,
Miles Cunningham, David Weingarten, Joseph D. Ciacci,6 Stefan Juhas, Jana
Juhasova, Jan Motlik,
Michael P. Hefferan, Tom Hazel, Karl Johe,Cassiano Carromeu, Alysson Muotri,
Jack Bui,
Jan Strnadel and Martin Marsala

http://pigmod.avcr.cz/miranda2/export/sitesavcr/pigmod/veda-a-
vyzkum/publikace/files/06_dolezalova_cne23575.pdf

2014

ABSTRACT
An important component for successful translation of cell replacement-based
therapies into clinical practice is the utilization of large animal models to conduct
efficacy and/or safety cell dosing studies. Over the past few decades, several large
animal models (dog, cat, nonhuman primate) were developed and employed in cell
replacement studies; however, none of these models appears to provide a readily
available platform to conduct effective and large-scale preclinical studies. In recent
years, numerous pig models of neurodegenerative disorders were developed using
both a transgenic approach as well as invasive surgical techniques. The pig model
(na€ıve noninjured animals) was recently used successfully to define the safety
and optimal dosing of human spinal stem cells after grafting into the central
nervous system (CNS) in immunosuppressed animals. The data from these studies
were used in the design of a human clinical protocol used in amyotrophic lateral
sclerosis (ALS) patients in a Phase I clinical trial. In addition, a highly inbred
(complete major histocompatibility complex [MHC] match) strain of miniature pigs
is available which permits the design of comparable MHC combinations between
the donor cells and the graft recipient as used in human patients. Jointly, these
studies show that the pig model can represent an effective large animal model to
be used in preclinical cell replacement modeling. This review summarizes the
available pig models of neurodegenerative disorders and the use of some of these
models in cell replacement studies. The challenges and potential future directions
in more effective use of the pig neurodegenerative models are also discussed.

11. Article

Mapping QTL in the porcine MHC region affecting fatness and

growth traits in a Meishan/Large White composite population

W. H. Wei, T. M. Skinner, J. A. Anderson, O. I. Southwood, G. Plastow, A. L.

Archibald and C. S. Haley.

http://www.animalgenome.org/QTLdb/references/20477798.pdf

2010

A number of studies have mapped QTL regulating porcine fatness and growth
traits to the region of the major histocompatibility complex (MHC) on porcine
chromosome 7 using various experimental crosses. The QTL results from crosses
using the Chinese Meishan (MS) (slow growing and fat) are particularly interesting
because the MS alleles have been found to be associated with increased growth
rate and reduced backfat depth. We investigated these QTL further in a composite
population derived previously over eight generations by intercrossing Meishan and
the European Large White breeds. Genotype information from 32 markers in a
15cM target region was used in linkage and association analyses. A twostep
variance component analysis identified QTL for three growth-related traits,
explaining 19 24% of the phenotypic variance with a confidence interval of 4 cM in
the target region. SNP association analyses found that ss181128966 and
ss181128924 within the QTL interval were strongly associated with the growth
traits. Only weak signals for an effect on backfat depth were found in the
association and linkage analyses, possibly because of past directional selection in
the composite population.

12. Article

Unique Characteristics of the Immune Systems in Ruminants and

Pigs
William C. Davis and Mary J. Hamilton

http://www.ars.usda.gov/sp2UserFiles/ad_hoc/36253000VirtualConference/Posters
/G00061.pdf

2010

ABSTRACT

Comparative analysis of the immune systems of food and companion animals has
revealed differences in the composition of T lymphocyte subpopulations in
ruminants and pigs. The difference in ruminants is attributable to the presence of a
major subset of gamma/delta T cells that appears to have undergone expansion in
the course of evolutionof ruminants. This subset is characterized by the expression
of two lineage restricted molecules, WC1 (bovine workshop cluster 1) andGD3.5
and restricted usage of gamma genes of TCR1 and Vgammasequences. The
difference in pigs is attributable to the presenceof a similar orthologous subset of
gamma/delta T cells and the expression ofCD4, CD8, and MHC class II molecules
on alpha/beta T cell subsets. As inruminants, the unique gamma/delta T cell subset
is characterizedby the expression of two lineage restricted molecules, one
thatappears to be the porcine orthologue of WC1 and WC6 (porcineworkshop
cluster 6). No information is available on the possiblerelation of GD3.5 and WC6.
In contrast to other species, alpha/betaT cells are divided into multiple subsets
characterized by thedifferential expression of CD4, CD8alpha/alpha,
CD8alpha/beta, andMHC class II molecules. These observations suggest the
mechanismsregulating the development and expression of immune responses
toinfectious agents and derived vaccines may differ from thosedescribed in rodents
and humans.

13. Article

Partial protection against classical swine fever virus elicited by

dendrimeric vaccine-candidate peptides in domestic pigs

Joan Tarradas, Marta Monsó, Marta Munoz, Rosa Rosell, Lorenzo Frailea, Maria
Teresa Frías, Mariano Domingoa, David Andreub, Francisco Sobrino, Llilianne
Ganges
 http://www.upf.edu/uprot/_pdf/205-Vaccine2011.pdf

2011

ABSTRACT

We report the immunogenicity of three dendrimeric peptide vaccine candidates for

classical swine fever virus (CSFV). Each dendrimeric construct contained four
copies of a B-cell epitope from the E2 glycoprotein of CSFV [construct 1: E2 (694–
712); 2: E2 (712–727); 3: E2 (829–842)] joined to a T-cell epitope from the NS3
protein (residues 1446–1460). Intramuscular immunization of domestic pigs with
the different constructs significantly reduced the clinical score after lethal challenge
with CSFV. In contrast, control pigs developed severe clinical signs of the disease.
All pigs vaccinated with construct 1, containing a B-cell epitope from the E2 B–C
domain, developed an antibody response that recognized not only the original
dendrimeric immunogen but also its constituting E2 epitope in linear form, albeit no
neutralizing antibodies were detected prior to viral challenge. Two of these pigs
were partially protected, which associated with the induction of IFN- producing cells
and of neutralizing antibodies upon challenge. Interestingly, the serological
response elicited by construct 1 lacked antibodies to E2 A domain, used as
infection markers. The dendrimeric approach could therefore provide a basis for
the development of CSFV marker (DIVA) vaccines, and contribute to a better
understanding of the immune responses against CSFV.

14. Article

Cellular immunity in ASFV responses

Haru-Hisa Takamatsua, Michael S. Denyer, Anna Lacasta, Catrina M.A. Stirlinga,

Jordi M. Argilaguet, Christopher L. Nethertona, Chris A.L. Ouraa, Carlos Martins,
Fernando Rodríguez

http://asf-referencelab.info/asf/images/files/INMUNIDAD_CLAR.pdf

2010

ABSTRACT

African swine fever virus (ASFV)infection usually results in an acute haemorrhagic

disease with a mortality rate approaching 100% in domestic pigs. However, pigs
can survive infection with less-virulentisolates of ASFV and may become
chronically infected. Surviving animals are resistant to challenge with homologous
or, in some cases, closely related isolates of the virus indicating that pigs can
develop protective immunity against ASFV. During asymptomatic, non-virulent
ASFV infections natural killer cell activity increases in pigs, suggesting this cell type
plays a role in ASFV immunity. Furthermore, depletion of CD8+ lymphocytes from
ASFV immune pigs demolishes protective immunity against related virulent
viruses. This suggests that ASFV specific antibody alone is not sufficient for
protection against ASFV infection and thatthere is an important role for the CD8+
lymphocyte subsetin ASFV protective immunity. These results were supported by
DNA immunization studies, demonstrating a correlation between the protection
afforded against lethal challenge and the detection of a large number of vaccine-
induced antigen-specific CD8+ T-cells. Peripheral blood mononuclear cells
(PBMCs) from ASF immune pigs protected from clinical disease show higher
proportions of ASFV specific CD4+CD8high+ double positive cytotoxic T cells than
PBMCs from ASF immune but clinically diseased pig. The frequency of ASFV
specific IFN producing T cells induced by immunization correlates to the degree of
protection from ASFV challenge, and this may prove to be a useful indicator of any
potential cross-protection against heterologous ASFV isolates.

15. Article

Creating Class I MHC−Null Pigs Using Guide RNA and the Cas9
Endonuclease

Luz M. Reyes, Jose L. Estrada, Zheng Yu Wang, Rachel J. Blosser, Rashod F.

Smith, Richard A. Sidner, Leela L. Paris, Ross L. Blankenship, Caitlin N. Ray,
Aaron C. Miner, Matthew Tector and A. Joseph Tector

http://www.jimmunol.org/content/early/2014/10/19/jimmunol.1402059.full.pdf+html

2014

ABSTRACT

Pigs are emerging as important large animal models for biomedical research, and
they may represent a source of organs for xenotransplantation. The MHC is pivotal
to the function of the immune system in health and disease, and it is particularly
important in infection and transplant rejection. Pigs deficient in class I MHC could
serve as important reagents to study viral immunity as well as allograft and
xenograft rejection. In this study, we report the creation and characterization of
class I MHC knockout pigs using the Cas9 nuclease and guide RNAs. Pig fetal
fibroblasts were genetically engineered using Cas9 and guide RNAs, and class I
MHC2 cells were then used as nuclear donors for somatic cell nuclear transfer. We
produced three piglets devoid of all cell surface class I proteins. Although these
animals have reduced levels of CD42CD8+ T cells in peripheral blood, the pigs
appear healthy and are developing normally. These pigs are a promising reagent
for immunological research.

16.Article
 Haplessly Hoping: Macaque Major Histocompatibility Complex
Made Easy
Roger W. Wiseman, Julie A. Karl, Patrick S. Bohn, Francesca A. Nimityongskul,
Gabriel J. Starrett, and David H. O’Connor

https://ehr.primate.wisc.edu/wiki/WNPRC/WNPRC_Units/Research_Services/Gene
tics_Services/Public/download.view?entityId=469a47b9-8451-102c-8dc0-
493dbd27fbc9&name=WisemanRW.ILAR.2013.pdf

2013

ABSTRACT

Major histocompatibility complex (MHC) gene products control the repertoire of T

cell responses that an individual may create against pathogens and foreign tissues.
This text will review the current understanding of MHC genetics in nonhuman
primates, with a focus on Mauritian-origin cynomolgus macaques (Macaca
fascicularis) and Indian-origin rhesus macaques (Macaca mulatta). These closely
related macaque species provide important experimental models for studies of
infectious disease pathogenesis, vaccine development, and transplantation
research. Recent advances resulting from the application of several cost effective,
highthroughput approaches, with deep sequencing technologies have
revolutionized our ability to perform MHC genotyping of large macaque cohorts.
Pyrosequencing of cDNA amplicons with a Roche/454 GS Junior instrument,
provides excellent resolution of MHC class I allelic variants with semi-quantitative
estimates of relative levels of transcript abundance. Introduction of the Illumina
MiSeq platform significantly increased the sample throughput, since the sample
loading workflow is considerably less labor intensive, and each instrument run
yields approximately 100-fold more sequence data. Extension of these sequencing
methods from cDNA to genomic DNA amplicons further streamlines the
experimental workflow and opened opportunities for retrospective MHC genotyping
of banked DNA samples. To facilitate the reporting of MHC genotypes, and
comparisons between groups of macaques, this text also introduces an intuitive
series of abbreviated rhesus MHC haplotype designations based on a major
Mamu-A or Mamu-B transcript characteristic for ancestral allele combinations. The
authors believe that the use of MHC-defined macaques promises to improve the
reproducibility, and predictability of results from pre-clinical studies for translation to
humans.

17. Article

Human dominant-negative class II transactivator transgenic pigs

– effect on the human anti-pig T-cell immune response and
immune status
Hidetaka Hara, William Witt, Tanner Crossley, Cassandra Long, Kumiko Isse, Liming Fan,
Carol J. Phelps, David Ayares, David K. C. Cooper, Yifan Dai and Thomas E. Starz.

http://onlinelibrary.wiley.com/store/10.1111/imm.12107/asset/imm12107.pdf?v=1&t=ijqmr
hb3&s=67c8dde091ba5ace72013710415a5f255f031eb7

2013

ABSTRACT

Swine leucocyte antigen (SLA) class II molecules on porcine (p) cells play a crucial
role in xenotransplantation as activators of recipient human CD4+ T cells. A human
dominant-negative mutant class II transactivator (CIITA-DN) transgene under a
CAG promoter with an endotheliumspecific Tie2 enhancer was constructed. CIITA-
DN transgenic pigs were produced by nuclear transfer/embryo transfer. CIITA-DN
pig cells were evaluated for expression of SLA class II with/without activation, and
the human CD4+ T-cell response to cells from CIITA-DN and wild-type (WT) pigs
was compared. Lymphocyte subset numbers and T-cell function in CIITA-DN pigs
were compared with those in WT pigs. The expression of SLA class II on antigen-
presenting cells from CIITA-DN pigs was significantly reduced (40–50% reduction
compared with WT; P < 001), and was completely suppressed on aortic endothelial
cells (AECs) even after activation (100% suppression; P < 001). The human CD4+
T-cell response to CIITA-DN pAECs was significantly weaker than to WT pAECs
(60–80% suppression; P < 001). Although there was a significantly lower frequency
of CD4+ cells in the PBMCs from CIITA-DN (20%) than from WT (30%) pigs (P <
001), T-cell proliferation was similar, suggesting no significant immunological
compromise. Organs and cells from CIITA-DN pigs should be partially protected
from the human cellular immune response.

18. Article

Feral pigs in the northern South Island, New Zealand: I. Origin,

distribution, and density
C. M. H. Clarke & R. M. Dzieciolowski

http://www.tandfonline.com/doi/abs/10.1080/03036758.1991.10418181

2012

ABSTRACT

Feral pigs in the northern South Island are derived from 14 breeds which originated
under domestication in Polynesia, Europe, and Asia. Pigs were first established in
the Marlborough Sounds by Captain Cook in 1773. The original animals were
obtained in Tonga and Tahiti, and were probably S. scrofa vittatus, of Indo-Malay
origin. The domestic breeds subsequently established between the 1830s and the
1970s were mostly of Eurasian origin. These breeds had diverse genetic histories
and physical characteristics. Feral pigs now occupy 30% (20, 000 km2) of the study
area. Overall distribution has expanded slightly (6%) since the 1970s because of
liberations and natural spread. Pig distribution by habitat was: farmland 56%;
indigenous forests 37%; and pine plantations 7%. Pig densities have declined
since the 1970s as a result of intensified hunting.

19. Article

A Two-Compartment Mathematical Model of Endotoxin-induced

Inflammatory and Physiologic Alterations in Swine
Gary Nieman, B.S.,David Brown, B.S.,Joydeep Sarkar, Ph.D.,Brian Kubiak,
M.D.,Cordelia Ziraldo, B.S.,Joyeeta Dutta-Moscato, M.S.,Christopher Vieau,
B.S.,Derek Barclay, B.S.,Louis Gatto, Ph.D.,Kristopher Maier, Ph.D.,Gregory
Constantine, Ph.D.,Timothy R. Billiar, M.D.,Ruben Zamora, Ph.D., Qi Mi,
Ph.D., Steve Chang, M.S.,and Yoram Vodovotz, Ph.D.

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3308118/

2012

ABSTRACT

Swine exhibited various degrees of inflammation and acute lung injury (ALI),
including one death with severe ALI (P/F ratio <200 and static compliance <10
L/cmH2O). Plasma interleukin (IL)-1β, IL-4, IL-6, IL-8, IL-10, tumor necrosis factor
(TNF)-α, high mobility group box-1 (HMGB1), and NO2−/NO3−,, were significantly (p
<0.05) elevated over the course of the experiment. Principal Component Analysis
(PCA) was used to suggest principal drivers of inflammation. Based in part on
PCA, an Ordinary Differential Equation (ODE) model was constructed, consisting
of the lung and the blood (as a surrogate for the rest of the body), in which
endotoxin induces TNF-αin monocytes in the blood, followed by the trafficking of
these cells into the lung leading to the release of HMGB1, which in turn stimulates
the release of IL-1βfrom resident macrophages. The ODE model also included
blood pressure, PaO2, and FiO2, and a damage variable that summarizes the
health of the animal. This ODE model could be fit to both inflammatory and
physiologic data in the individual swine. The predicted time course of damage
could be matched to the Oxygen Index in 3 of the 4 swine.
20. Article

Specific hunger- and satiety-induced tuning of guinea pig enteric

nerve activity
Lina Roosen,Werend Boesmans,Marjan Dondeyne,Inge Depoortere,Jan
Tack,Pieter Vanden Berghe

http://onlinelibrary.wiley.com/doi/10.1113/jphysiol.2012.231134/full

2012

ABSTRACT

Although hunger and satiety are mainly centrally regulated, there is convincing
evidence that also gastrointestinal motor activity and hormone fluctuations
significantly contribute to appetite signalling. In this study, we investigated how
motility and enteric nerve activity are set by fasting and feeding. By means of
video-imaging, we tested whether peristaltic activity differs in ex vivo preparations
from fasted and re-fed guinea pigs. Ca2+ imaging was used to investigate whether
the feeding state directly alters neuronal activity, either occurring spontaneously or
evoked by (an)orexigenic signalling molecules. We found that pressure-induced (2
cmH2O) peristaltic activity occurs at a higher frequency in ileal segments from re-
fed animals (re-fed versus fasted, 6.12 ± 0.22 vs. 4.84 ± 0.52 waves min−1, P=
0.028), even in vitro hours after death. Myenteric neuronal responses were tuned
to the feeding status, since neurons in tissues from re-fed animals remained hyper-
responsive to high K+-evoked depolarization (P < 0.001) and anorexigenic
molecules (P < 0.001), while being less responsive to orexigenic ghrelin (P=
0.013). This illustrates that the feeding status remains ‘imprinted’ex vivo. We were
able to reproduce this feeding state-related memory in vitro and found humoral
feeding state-related factors to be implicated. Although the molecular link with
hyperactivity is not entirely elucidated yet, glucose-dependent pathways are clearly
involved in tuning neuronal excitability. We conclude that a bistable memory
system that tunes neuronal responses to fasting and re-feeding is present in the
enteric nervous system, increasing responses to depolarization and anorexigenic
molecules in the re-fed state, while decreasing responses to orexigenic ghrelin.
Unlike the hypothalamus, where specific cell populations sensitive to either
orexigenic or anorexigenic molecules exist, the enteric feeding state-related
memory system is present at the functional level of receptor signalling rather than
confined to specific neuron subtypes.
21. Article

Development of a new auxiliary heterotopic partial liver

transplantation technique using a liver cirrhosis model in
minipigs: Preliminary report of eight transplants
Un-Jing Zhang,Jian-Xiang Niu,Gen-Quan Yue,Hai-Yan Zhong,Xing-Kai Meng

http://www.spandidos-publications.com/10.3892/etm.2012.507

2012

ABSTRACT

This study aimed to develop a new auxiliary heterotopic partial liver transplantation
(AHPLT) technique in minipigs using a model of liver cirrhosis. Based on our
previous study, 14 minipigs were induced to cirrhosis by administration of carbon
tetrachloride (CCl4) through intraperitoneal injection. All of the cirrhotic animals
were utilized as recipients. The donor's liver was placed on the recipient's splenic
bed, and the anastomosis was performed as follows: end-to-end anastomosis
between the donor's portal vein and the recipient's splenic vein, end-to-side
anastomosis between the donor's suprahepatic vena cava and the recipient's
suprahepatic vena cava, and end-to-end anastomosis between the donor's hepatic
artery and the recipient's splenic artery. The common bile duct of the donor was
intubated and bile was collected with an extracorporeal bag. Vital signs, portal vein
pressure (PVP), hepatic venous pressure (HVP) and portal vein pressure gradient
(PVPG) were monitored throughout the transplantation. All 8 minipigs that
developed liver cirrhosis were utilized to establish the new AHPLT; 7 cases
survived. Following the surgical intervention, the PVP and PVPG of the recipients
were lower than those prior to the operation (P<0.05), whereas the PVP and PVPG
of the donors increased significantly compared to those of the normal animals
(P<0.05). A new operative technique for AHPLT has been successfully described
herein using a model of liver cirrhosis.

22. Article

Airways dilate to simulated inspiratory but not expiratory manoeuvres

Adrian R. West, Elangovan Thaya Needi, Howard W. Mitchell, Peter

K. McFawn, Peter B. Noble

http://erj.ersjournals.com/content/40/2/455

2010
 ABSTRACT

In a healthy human, deep inspirations produce bronchodilation of contracted

airways, which probably occurs due to the transient distension of the airway
smooth muscle (ASM). We hypothesised that deep expiratory manoeuvres also
produce bronchodilation due to transient airway wall and ASM compression.

We used porcine bronchial segments to assess the effects of deep inspirations,

and maximal and partial expiration (submaximal) on airway calibre. Respiratory
manoeuvres were simulated by varying transmural pressure using a hydrostatic
pressure column: deep inspiration 5 to 30 cmH2O, maximal expiration 30 to -15
cmH2O, partial expiration 10 to -15 cmH2O; amidst a background of tidal
oscillations, 5 to 10 cmH2O at 0.25 Hz. Changes in luminal cross-sectional area in
carbachol-contracted airways were measured using video endoscopy.
Deep inspirations produce an immediate bronchodilation (∼40–60%, p=0.0076)
that lasts for up to 1 min (p=0.0479). In comparison, after maximal expiration there
was no immediate change in airway calibre; however, a delayed bronchodilatory
response was observed from 4 s after the manoeuvre (p=0.0059) and persisted for
up to 3 min (p=0.0182). Partial expiration had little or no effect or airway calibre.

The results observed demonstrate that the airway wall dilates to deep inspiration
manoeuvres but is unresponsive to deep expiratory manoeuvres.

23. Article

A Pig Model of the Preterm Neonate: Anthropometric and

Physiological Characteristics

Yvonne A. Eiby ,Layne L. Wright,Viskasari P. Kalanjati,Stephanie M. Miller,Stella

T. Bjorkman,Helen L. Keates,Eugenie R. Lumbers,Paul B. Colditz,Barbara E.
Lingwood

http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0068763#abstract0

2013

ABSTRACT
Large animal models are an essential tool in the development of rationally-based
new clinical therapies for preterm infants. We provide a description of the newborn
pig as a model of the preterm neonate in terms of growth parameters, physiology
and the requirement for intensive care over a range of gestational ages.
Twenty-nine litters of piglets (n = 298) were delivered by caesarean section at six
timepoints during gestation from 91d to 113d (term = 115d). Two groups, at 91 and
97d gestation, also received maternal glucocorticoid treatment. At four of these
timepoints, piglets (n = 79) were ventilated, sedated and monitored using standard
neonatal intensive care techniques for up to 8 h in various experimental protocols.

24. Article

Four-dimensional visualization of subpleural alveolar dynamics in

vivo during uninterrupted mechanical ventilation of living swine
Eman Namati, William C. Warger, Carolin I. Unglert, Jocelyn E. Eckert, Jeroen
Hostens, Brett E. Bouma, and Guillermo J. Tearney

https://www.osapublishing.org/boe/fulltext.cfm?uri=boe-4-11-
2492&id=269019

2013

ABSTRACT

Pulmonary alveoli have been studied for many years, yet no unifying hypothesis
exists for their dynamic mechanics during respiration due to their miniature size
(100-300 μm dimater in humans) and constant motion, which prevent standard
imaging techniques from visualizing four-dimensional dynamics of individual
alveoli in vivo. Here we report a new platform to image the first layer of air-filled
subpleural alveoli through the use of a lightweight optical frequency domain
imaging (OFDI) probe that can be placed upon the pleura to move with the lung
over the complete range of respiratory motion. This device enables in-
vivo acquisition of four-dimensional microscopic images of alveolar airspaces
(alveoli and ducts), within the same field of view, during continuous ventilation
without restricting the motion or modifying the structure of the alveoli. Results from
an exploratory study including three live swine suggest that subpleural alveolar air
spaces are best fit with a uniform expansion (r2 = 0.98) over a recruitment model
(r2 = 0.72). Simultaneously, however, the percentage change in volume shows
heterogeneous alveolar expansion within just a 1 mm x 1 mm field of view. These
results signify the importance of four-dimensional imaging tools, such as the device
presented here. Quantification of the dynamic response of the lung during
ventilation may help create more accurate modeling techniques and move toward a
more complete understanding of alveolar mechanics.

25. Article

Effects of breed, postnatal development, and nutrition on mRNA

expression of the FTO gene in porcine muscle and its relationship
with intramuscular fat deposition

X. Tao, X.M. Men, B. Deng, Z.W. Xu

http://81.0.228.28/publicFiles/97599.pdf

2013

ABSTRACT

The effects of breed, development, and nutrition on mRNA expression of the fat
mass and obesityassociated gene (FTO) and its relationship with intramuscular fat
(IMF) content in porcine muscle (m. longissimus dorsi; m.l.d.) were estimated.
Purebred Jinhua, Zhongbai, Yorkshire, Duroc, Duroc × Zhongbai (DZ), and Duroc
× Yorkshire × Landrace (DYL) pigs were used to investigate the effect of breed.
Pigs weighing 2.5, 10, 20, 40, 60, and 100 kg were selected to study the effects of
different stages of development. To study the effect of nutrition, four diets were
selected: corn-soybean (CS), CS with 1.2% conjugated linoleic acid (CLA) or
0.05% creatine monohydrate (CMH), and barley-soybean (BS). All eighty animals
were slaughtered, and m.l.d. samples were collected to examine FTO mRNA
expression and IMF content. Results showed that breed significantly affected FTO
mRNA expression and IMF content. FTO mRNA expression in the studied pigs
was in the order: Zhongbai and Yorkshire > Duroc and DZ > Jinhua and DYL. The
IMF content ordered by breed was Duroc > DZ > DYL > Jinhua > Zhongbai >
Yorkshire. Both FTO mRNA expression and IMF content increased with age of the
pigs, with the greatest difference seen between 100 kg pigs and all other weights.
In the study, none of the four diets had a significant effect (P > 0.05) on FTO
mRNA expression or IMF content. The study demonstrated that FTO mRNA
expression increased with increasing body weight and was significantly affected by
the breed of pigs. The results showed that FTO mRNA expression had an
inconsistent correlation with IMF content between breeds and developmental ages.