Cross Reactivity against COVID-19 IgG

Dr. Nayab Zehra

PGR Chemical Pathology
 Antigen
Antigen is a molecule present at the outside of a pathogen or
human cell, that can be bound to by an antibody. The presence of
antigens in the body normally triggers an immune response.

Common antigens include microorganisms like viruses, fungi, and

bacteria and toxins like pesticide residue and heavy metals. These
external antigens are known as foreign antigens because they
originate outside of the body. There are also autoantigens, or self-
antigens, which are substances produced inside of the body that
prompt an immune response
 Epitope
• Epitope, also known as antigenic determinant, is the part of an antigen
that is recognized by the immune system, specifically by antibodies, B
cells, or T cells.
• It is the specific piece of the antigen to which an antibody binds.
• An antigen may contain more than one type of such molecule.
• The number of epitopes varies with size and complexity of structure.
• The effective number of epitopes on an antigen is its Valency
 Haptens
Small molecule which, when combined with a larger carrier
such as a protein, can elicit the production of antibodies e.g.
monosaccharides, peptides hormones and drugs e.g penicillin.
 Antibody
• Antibody, also known as an immunoglobulin, is a large, Y-shaped
protein produced mainly by plasma cells that is used by the
immune system to neutralize pathogens or act against self
antigens.
• Antibodies are created by plasma cells which are derived from B
lymphocytes.
• Antibodies recognize the 3 dimensional shape of an epitope as
well as its basic amino acid structure.
Antibody Structure
 Paratope
• Paratope, also called an antigen-binding site, is a part of an antibody
which recognizes and binds to an antigen.
• It is a small fragment of antibody's Fab region, composed of heavy
and light chains parts.
• It combines with an epitope in Lock-and-Key manner and affinity of
an antibody for an antigenic determinant is directly proportional to
closeness of fit.
Epitope & Paratope
Structure of different Antibodies
Characteristics of Antibody Types
 Affinity
• Antibody affinity is the strength with which antibody binds to its target antigen.
• It measures the strength of interaction between an epitope and an antibody’s
binding site.
• It is influenced by a variety of factors including valency and the precise fit of
the antigen in the antigen binding groove. 
• When an antigen is encountered for the first time, the affinity of the antibodies
produced is low. Once the body is familiar with the antigen, the immune
response adapts and the binding affinity increases.
 Avidity
• It is measure of the total binding strength of an antibody on all
binding sites.
• Whilst affinity represents the binding strength between one paratope
and one epitope
• It depends on the following:
1) The binding affinity
2) The valency
3) The structural arrangement
 Heterogeneity
Antibodies can bind to different configurations of a
single antigenic determinant. Thus different antibody
molecules can be produced against such antigens .
Heterogeneity
(Cont’d)
 Cross Reactivity
• The ability of an antibody to react with similar
antigenic sites on different proteins.
• It is by-product of heterogeneity on an
immune response.
• It leads to false positive results.
 Cross Reactivity (Cont’d)

• It may provide cross-protective immunity to related pathogens.

• It allows the same antibody to recognize and provoke a similarly robust
immune response to a closely related but previously unencountered
antigen. In this way, a less specific antibody provides broad-spectrum
immunity despite antigenic variation in pathogens.

• https://www.jove.com/science-education/10900/cross-reactivity
 Cross Reactivity (Cont’d)

• Cross-reactivity may be caused by identical carbohydrate structures on unrelated

proteins from the same or different species. Such 
cross-reactive carbohydrate determinants (CCDs) are an issue in allergy diagnosis
• It may increase in the number of allergens that an allergic person reacts to. While
the initial immune reaction was induced against a specific antigen, cross-
reactivity lets the immune system act against additional, structurally similar
antigens which can produce a hyperimmune reaction.

• https://en.wikipedia.org/w/index.php?title=Cross-reactivity&oldid=970313377
 Cross Reactivity
(COVID-19 & Autoimmune Disorders)
• In a study carried out in China , total 290 serum samples were collected,
consisting of 98 patients with rheumatoid arthritis, 100 patients with
systemic lupus erythematosus, and 92 patients with Sjogren’s
syndrome. The samples collected predates the COVID-19 pandemic.
• The results showed that both IgG and IgM antibodies against SARSCoV-2
were not detected in the serum of patients with autoimmune disease,
indicating that there was no cross reactivity between autoantibodies and
SARS-CoV-2 antibodies

• https://doi.org/10.1016/S2665-9913(20)30128-4
 Cross Reactivity
(COVID-19 & Dengue)
• In a study carried out in Singapore, total of 44 anonymized DENV-
positive sera, from cases of acute dengue (IgM and IgG positivity),
were tested by using COVID-19 IgG/ IgM Rapid Test Cassette.
• The result showed cross reactivity of 2 samples among 44 DENV-
positive sera tested for COVID-19 antibodies.

Sethuraman N, Jeremiah SS, Ryo A. Interpreting diagnostic tests for SARS-CoV-2. JAMA. 2020;
10.1001/jama.2020.8259. [published online ahead of print].
 Cross Reactivity
(COVID-19 & Granulomatosis with Polyangiitis)

• An 82-year-old female, who was admitted to hospital with symptoms of fever and
general fatigue that had lasted 7 days. She already had a positive IgM for COVID-19.In the
following days, she deteriorated. Later on her immunologic profile revealed positive c-
ANCA . Treatment started immediately. At day 10, treatment response was noticed as
indicated by respiratory and renal function improvement. This report highlights the need
for meticulous patient evaluation in order to avoid misdiagnosis in the era of pandemic.
• It was suggested in the study that patients with autoimmune diseases, including ANCA-positive-
vasculitis, present with a plethora of autoantibodies, and SARS-CoV lysates used as antigens could
have led to such false-positive reactions.

• https://www.frontiersin.org/article/10.3389/fmed.2020.00399
 Cross Reactivity
(COVID-19 & SLE)
• 104 serum specimens from SLE patients were collected.
• 58 (32.8%) showed cross reactivity with SARS-CoV-IgG antibodies
• This study showed that SARS-CoV lysates for the ELISA to detect SARS-
CoV antibodies could lead to the false-positive reactions or cross-
reactions of SARS-CoV antibodies in non-SARS diseases.

Wang Y, Sun S, Shen H, et al. Cross-reaction of SARS-CoV antigen with autoantibodies in

autoimmune diseases. Cellular & Molecular Immunology. 2004 Aug;1(4):304-307.
Manufacturer’s Claim (Abbott)
Manufacturer’s Claim (Cont’d)

The manufacturer (Abbott) claimed that out of 18

Infectious & 05 Autoimmune disease categories. Only 01
case of CMV showed cross reactivity out of 112 total
samples
Research Project

Cross Reactivity of COVID-19

IgG with known Immune
Mediated Disorders
 Material & Methods
• Study type: Cross sectional study
• Duration: 15 July - 30 September 2020
• Samples: Total 115 samples of following diseases:
• Typhoid, Hep A , Hep B , Hep C , Hep D, Hep E, ds DNA , Syphilis , ANA positive ,
Varicella Zoster , Rabies, Toxoplasmosis , EBV , Rubella, Rheumatoid Arthritis, HIV ,
Dengue
• 03 samples of healthy multiparous women were also included
• Statistical Analysis: SPSS 23.0 was used for statistical analysis.
Univariate analysis was done to find out percentages and frequencies.
 Analytical Technique
Equipment : Chemistry Autoanalyzer (Alinity I - Abbott) by
using Chemiluiminescence immunoassay technique
Reagents : Purified SARS-CoV-2 recombinant antigen coated
microparticles in TRIS buffer with surfactant.
Anti-human IgG (mouse, monoclonal) conjugate in MES buffer with
surfactant and protein (bovine) stabilizer
 Results
Serial Disease Number of COVID-19 IgG COVID-19 IgG
No. Category cases ( n) Reactive Non reactive
1 Typhoid 15 08 07
2 Hep A 05 0 05
3 Hep B 04 0 04
4 Hep C 16 02 14
5 Hep D 08 0 08
6 Hep E 14 0 14
7 Syphillis 05 01 04
8 Rabies 06 0 06
9 HIV 07 0 07
10 Varicella Zoster 01 0 01
Serial Disease Number of COVID-19 IgG COVID-19 IgG
No. Category Cases (n) Reactive Non Reactive

11 ANA 05 0 05

12 dsDNA 08 0 0
13 Rubella 03 0 03
14 Toxoplasma 03 0 03
15 Rheumatoid 08 0 08
Arthritis
16 Beta hCG 03 0 03
17 EBV 02 0 02
18 Dengue 02 0 02
 Findings
• Out of total 115 samples, 11 samples showed cross reactivity.
• Typhoid showed the highest number (08 samples) of cross reactivity
• Hepatitis C showed cross reactivity in 02 samples
• Syphilis showed cross reactivity in 01 sample
• Autoimmune disorders (included in study) revealed no cross reactivity
 Current Status
• Research work completed in all aspects
• Manuscript writing in process and near to
completion
• Ready to submit for publication in Pakistan
Armed Forces Medical Journal
 Conclusion
• Affinity , Avidity & Heterogeneity of an antibody play an
important role in cross reactivity.
• Manufacturer’s claim should not be accepted blindly and
must be verified by the lab.
• Any novel disease e.g COVID-19 should be aggressively
addressed for cross reactivity against common
autoimmune and infectious diseases.