Journal of Immunoassay and Immunochemistry

ISSN: 1532-1819 (Print) 1532-4230 (Online) Journal homepage: https://www.tandfonline.com/loi/ljii20

Singlet oxygen species and systemic lupus

erythematosus: a brief review

Rizwan Ahmad & Haseeb Ahsan

To cite this article: Rizwan Ahmad & Haseeb Ahsan (2019) Singlet oxygen species and systemic
lupus erythematosus: a brief review, Journal of Immunoassay and Immunochemistry, 40:4,
343-349, DOI: 10.1080/15321819.2019.1616555

To link to this article: https://doi.org/10.1080/15321819.2019.1616555

Published online: 22 May 2019.

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JOURNAL OF IMMUNOASSAY AND IMMUNOCHEMISTRY
2019, VOL. 40, NO. 4, 343–349
https://doi.org/10.1080/15321819.2019.1616555

Singlet oxygen species and systemic lupus erythematosus:

a brief review
Rizwan Ahmada and Haseeb Ahsan b

a
College of Medicine, Imam Abdulrahman bin Faisal University, Dammam, Saudi Arabia; bDepartment
of Biochemistry, Faculty of Dentistry, Jamia Millia Islamia, New Delhi, India

ABSTRACT KEYWORDS
Reactive oxygen species (ROS) are implicated in inflammatory, Singlet oxygen; anti-DNA
autoimmune, and neurodegenerative diseases. The activation, autoantibodies;
proliferation, and apoptosis of immune cells are dependent on autoimmunity; SLE
the controlled production and elimination of ROS. However,
under chronic inflammatory conditions, large amounts of ROS
generated are a major cause of many human diseases. The elec-
tronically excited molecular oxygen species known as singlet
oxygen (1O2) is a form of ROS and is one of the major cytotoxic
species in eukaryotic cells. ROS are known to cause DNA damage
leading to strand breaks, base damage, and conformational
changes. The 1O2, being one of the most potent ROS, is generated
by photoexcitation or by chemiexcitation and is known to selec-
tively react with the deoxyguanosine moiety in DNA. The biolo-
gical consequences of 1O2-induced damage causes loss of
transforming activity as well as mutagenicity and genotoxicity
and may lead to the formation of neo-epitopes in native DNA and
generation of pathogenic anti-DNA antibodies. The excessive
production of ROS may be one of the factors responsible for the
induction of autoimmune response in diseases such as cancer
and systemic lupus erythematosus.
Abbreviations: Singlet oxygen: 1O2; Superoxide anion: O2-.;
Hydroxyl radical: OH; Hydrogen peroxide: H2O2

Introduction
Free radicals are highly reactive ions and chemicals containing atoms with an
unpaired electron. The specific characteristics of these reactive molecules are
that they are highly unstable, short lived, and react with other stable mole-
cules to gain stability. They are synthesized as by-product of cellular respira-
tion and enzyme systems such as phagocytic cells, neutrophils, and
macrophages (NADPH oxidase, myeloperoxidases). The exposure to ionizing
radiation, smoke, herbicides, pesticides, fried foods, etc. may also lead to
their formation. These radicals react with important biomolecules such as
DNA, proteins, and lipids and may also lead to certain pathophysiological
conditions such as autoimmunity, cancer, and neurodegenerative diseases.

CONTACT Rizwan Ahmad [email protected]; [email protected] College of Medicine, Imam

Abdulrahman Bin Faisal University, Office #2011, Dammam, Saudi Arabia
© 2019 Taylor & Francis
344 R. AHMAD AND H. AHSAN

Although, the free radicals are sometimes harmful but they are also essential
to the normal cellular functions such as production of hormones (e.g.,
thyroxine), removing harmful bacteria and pathogens and for various
other physiological functions of the body.[1–4]
Reactive oxygen species (ROS) or oxygen free radicals are the chemical species
that have a single unpaired electron in the outer most orbit shell and are
generated as a product of oxygen degradation.[5] During normal respiration,
molecular oxygen is sequentially reduced by the addition of four electrons to
generate water. In this process number of toxic intermediates may be produced
which include superoxide anion radical (O2-.), hydrogen peroxide (H2O2), and
hydroxyl ion or radical (OH.). Therefore, the free radicals derived from mole-
cular oxygen represent the most important class of reactive species generated in
living systems.[6] The addition of one electron to dioxygen forms the O2-., arising
either through metabolic processes or oxygen “activation” by physical radiation,
and is considered the “primary” ROS which can further interact with other
molecules to generate “secondary” ROS either directly or through enzymatic or
metal catalyzed processes.[6,7] The O2-., which occurs intracellularly mostly
within the mitochondria during energy transduction through a small electron
“leak” to molecular oxygen, has been indicated in the pathophysiology of
a variety of diseases.[7] The OH., a neutral form of hydroxide ion (OH−), is
highly reactive, making it a dangerous radical with a very short in vivo half-life of
about 10−9 s. Additionally, reactive free radicals derived from oxygen that can be
formed in living systems are the peroxyl radicals (ROO). The simplest proto-
nated form is a conjugate acid of O2-. and is usually termed either the hydro-
peroxyl radical or perhydroxyl radical (HO2, HO2−).[7,8]
In living organisms, free radicals may be formed by exogenous interaction or
by physiological processes i.e. endogenous sources.[9] The exogenous sources of
ROS include electromagnetic and ultraviolet radiation, vehicle exhaust, cigarette
smoke, etc. The endogenous sources of ROS are mitochondrial electron trans-
port chain, respiratory burst of phagocytes, oxidation of lipids, etc. Excessive
ROS can induce oxidative stress and damage in cells and promote various
diseases such as degenerative diseases and aging.[10,11] Free radicals generated
by normal processes do not become harmful if a balance between their produc-
tion and removal/inactivation is maintained through quenching or antioxidants.
Antioxidants such as vitamin E (tocopherol, tocotrienol), superoxide dismutase
(SOD), catalase (CAT), glutathione peroxidase (GPx), vitamin C (ascorbic acid),
beta-carotene, coenzyme Q10 (CoQ10) provide electrons to free radicals to
neutralize them since these compounds are able to manage the loss of electron(s)
without becoming reactive. Therefore, these compounds have an essential role in
the quenching or removal of harmful free radicals.[2,12,13] Polyphenolic antiox-
idants can scavenge and reduce concentration of ROS and may prevent such
deleterious oxidative effects. The consumption or ingestion of pigments capable
 JOURNAL OF IMMUNOASSAY AND IMMUNOCHEMISTRY 345

of producing 1O2 after activation by sunlight can lead to severe photosensitivity

of skin (photosensitivity, photodermatitis, phytophotodermatitis).

Singlet oxygen
Singlet oxygen (dioxidene or dioxygen) is an inorganic chemical in an excited
state, with the chemical formula 1O2 (a1Δg, O2*). It is an electronically excited
state of molecular oxygen (O2), which is less stable than the normal triplet
oxygen and differs in their chemical properties.[14] Because of its unusual
properties, 1O2 can persist for over an hour at room temperature in isolation.
Singlet oxygen is highly reactive and is usually generated with a photosensitizer
pigment. The damaging effects of sunlight on many organic materials (polymers,
porphyrins, etc.) are often attributed to the effects of 1O2. The 1O2 is the active
species in photodynamic therapy and it is used to control and destroy cancer
cells.[15,16] In photosynthesis, 1O2 can be produced from the light harvesting
chlorophyll molecules. One of the function of carotenoids in photosynthetic
systems is to prevent damage caused by 1O2 either through the removal of excess
light photons or directly quenching 1O2. One of the roles of carotenoid in
photosynthetic systems is to prevent damage caused 1O2 by either removing
excess light energy from chlorophyll molecules or quenching 1O2 directly.[17] In
mammalian biology, 1O2 is one of the ROS which is linked to oxidation of LDL
cholesterol and the resultant cardiovascular effects.[18] Among the ROS, 1O2 is
formed under conditions of oxidative stress simultaneously with or as
a consequence of radical formation.[18–20] Furthermore, photosensitizes such
as riboflavin (vitamin B2), ethylene blue, rose bengal (dichlorotetraiodo fluor-
escein) in the presence of light generate 1O2.[21–23]
The 1O2 species was first observed in 1924 and defined as a more reactive form
of oxygen.[24–26] The 1O2 is the lowest excited state of the dioxygen molecule and its
main method of production is by photosensitization reaction.[27] Kautsky (1931)
first proposed that 1O2 might be a reaction intermediate in dye-sensitized photo-
oxygenation.[28] Its lifetime in solution is in the microsecond range (3 µs in water to
about 700 µ in C6D6).[29–33] Since molecular oxygen is ubiquitous and efficiently
quenches electronically excited states, 1O2 is likely to be formed following irradia-
tion in various chemical and biological processes as well as in several diseases.[26]

Singlet oxygen in systemic lupus erythematosus

Singlet oxygen has been implicated in various pathophysiological processes that
may lead to damage of biomolecules such as DNA, RNA, and proteins. DNA is
one of important targets of 1O2 leading to the loss of transforming activity,
mutagenicity, and genotoxicity.[34] The DNA modifications produced by 1O2 are
almost exclusively oxidized purines forming neo-epitopes on DNA and may lead
to the formation of autoantibodies. The native mammalian DNA (nDNA) per se
346 R. AHMAD AND H. AHSAN

is non-immunogenic, whereas the bacterial DNA is a stronger immunogen. The

immunogenicity of bacterial DNA has been attributed to the presence of nucleo-
tide hexamers containing unmethylated CpG motifs.[35] Similarly, the role of
unmethylated CpG dinucleotide sequences in the immunogenicity of plasmid
DNA is well recognized. Other modified forms of polynucleotides, nDNA, and
proteins have been reported to be immunogenic and the autoantibodies thus
generated cross-react with nDNA. Even though nucleic acid antigens are by
themselves poorly immunogenic, their antigenicity can be enhanced by mod-
ification with ROS.[4] The 1O2-modified plasmid DNA was found to be a potent
immunogen when immunized in rabbits inducing high titer antibodies. The
exposure to 1O2 might have generated potential neo-epitopes against which the
antibodies were raised. The data indicated a higher specificity of immune
immunoglobulin G toward 1O2-modified epitopes.[35–37] It can therefore be
postulated that in chronic inflammatory diseases, ROS production by phagocytic
cells may cause damage to DNA generating neo-epitopes leading to the produc-
tion of antibodies cross-reacting with nDNA. The binding of induced antibodies
against 1O2-modified plasmid DNA, with an array of nucleic acids antigens,
demonstrates their heterogeneous antigen binding characteristics. It can be said
that systemic lupus erythematosus (SLE) anti-DNA autoantibodies exhibit poly-
specificity with respect to antigen binding.[36–38]
It appears that a defect in the immune system plays an initiating role in the
autoimmune response seen in various autoimmune diseases such as SLE and
rheumatoid arthritis (RA) patients. Increased levels of circulating antibodies
and autoantibodies have also been reported in the serum of patients with
malignancies.[35] SLE is a multisystem autoimmune disease characterized by
various immunological disorders, including production of autoantibodies,
formation of immune complexes, decreased serum complement components,
and lymphocytopenia.[37] It has been reported that damage to nDNA by ROS
results in an increased binding of anti-DNA antibodies present in the sera of
SLE patients. The presence of these anti-DNA autoantibodies has long been
considered as a biomarker of the disease process and progress. Hence, the
ROS-modified DNA is a better and more discriminating immunogen than
nDNA for the production of anti-DNA autoantibodies in SLE. The detection
of 8-hydroxydeoxyguanosine, a biomarker of free radical mediated DNA
damage, in the sera of SLE patients demonstrates that ROS-modified DNA
may be involved in the pathogenesis of this autoimmune disorder. Thus, it
appears that ROS modification generates neo-epitopes on DNA that are
recognized by circulating anti-DNA autoantibodies. Anti-DNA antibodies
may bind with circulating 1O2-modified DNA antigens and contribute to
the formation of immune complexes in circulation leading to their deposition
in renal glomeruli. The preferential binding of SLE autoantibodies to mod-
ified DNA as compared to nDNA in enzyme-linked immunosorbent assay
indicates that O2-damaged/-modified DNA is highly immunogenic.[36,38]
 JOURNAL OF IMMUNOASSAY AND IMMUNOCHEMISTRY 347

Conclusion
Activation, proliferation, and apoptosis of immune cells are dependent on the
controlled production of ROS. However, under chronic inflammatory conditions,
large amounts of ROS generated are responsible for the pathophysiology of many
human disorders. Increased apoptosis and decreased clearance of apoptotic cells
observed in systemic autoimmunity might well be a contributory factor in auto-
immune disorders such as SLE and RA. ROS are known to cause DNA damage,
leading to strand breaks, base damage, and conformational changes. 1O2 is one of
the most potent ROS known to selectively react with the deoxyguanosine moiety
in DNA and may lead to the generation of neo-epitopes. Studies have shown that
the ROS-modified DNA is found in the sera of SLE and cancer patients. Hence,
the 1O2-modified DNA may be one of the factors responsible for the induction of
autoimmune response in SLE.

Acknowledgments
RA is grateful to Prof. Ali Ibrahim Al-Sultan, Dean, College of Medicine, IAU for the interest
shown in this study and the preparation of this manuscript. RA would also like to thank Vice
Deanship of Academic affairs for the necessary help and support. RA would like to thank the
Vice Deanship of Academic Affairs for the help and support and Prof. Saadat Ali, India for
his keen interest in the present study.

ORCID
Haseeb Ahsan http://orcid.org/0000-0002-5313-5959

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