Development and characterization of a panel of cross-reactive monoclonal antibodies generated using H1N1 influenza virus

Abstract

To characterize the antigenic epitopes of the hemagglutinin (HA) protein of H1N1 influenza virus, a panel consisting of 84 clones of murine monoclonal antibodies (mAbs) were generated using the HA proteins from the 2009 pandemic H1N1 vaccine lysate and the seasonal influenza H1N1(A1) vaccines. Thirty-three (39%) of the 84 mAbs were found to be strain-specific, and 6 (7%) of the 84 mAbs were subtype-specific. Twenty (24%) of the 84 mAbs recognized the common HA epitopes shared by 2009 pandemic H1N1, seasonal A1 (H1N1), and A3 (H3N2) influenza viruses. Twenty-five of the 84 clones recognized the common HA epitopes shared by the 2009 pandemic H1N1, seasonal A1 (H1N1) and A3 (H3N2) human influenza viruses, and H5N1 and H9N2 avian influenza viruses. We found that of the 16 (19%) clones of the 84 mAbs panel that were cross-reactive with human respiratory pathogens, 15 were made using the HA of the seasonal A1 (H1N1) virus and 1 was made using the HA of the 2009 pandemic H1N1 influenza virus. Immunohistochemical analysis of the tissue microarray (TMA) showed that 4 of the 84 mAb clones cross-reacted with human tissue (brain and pancreas). Our results indicated that the influenza virus HA antigenic epitopes not only induce type-, subtype-, and strain-specific monoclonal antibodies against influenza A virus but also cross-reactive monoclonal antibodies against human tissues. Further investigations of these cross-reactive (heterophilic) epitopes may significantly improve our understanding of viral antigenic variation, epidemics, pathophysiologic mechanisms, and adverse effects of influenza vaccines.

False‐positive semiquantitative immunochromatography assays for procalcitonin in three patients with rheumatoid arthritis—A case series

“…7 Heterophilic antibody interference has been reported for many tumor markers (alpha-fetoprotein, CA125, beta-human chorionic gonadotropin, squamous cell carcinoma antigen, and prostate-specific antigen), hormones (thyrotrophic hormone and estradiol), and infection markers (influenza and human hepatitis B antigen) using immunochromatography assays. 6,[8][9][10] Commercial quantitative methods have been improved to reduce heterophilic interference by removing the Fc portion and avoid the binding of heterophilic antibodies; this can also be achieved by adding a HAMA blocker or adsorbing the heterophilic antibodies using IgG from the same animal or serum, as a secondary antibody. In comparison, most immunochromatography assays, including PCT-Q ® , use whole immunoglobulins, which result in more frequent heterophilic interference.…”

Monoclonal antibody against H1N1 influenza virus hemagglutinin cross reacts with hnRNPA1 and hnRNPA2/B1

“…mAbs against the H1N1 influenza virus HA protein, including H1-84mAb, were prepared in our laboratory. The titre of the antibody was determined using the indirect ELISA method, and the reactivity of the antibody with the HA antigen was determined by western blotting (8). It has been previously determined that H1-84mAb binds to a nine-peptide linear epitope (191-LVLWGIHHP-199) on HA (12).…”

“…In our previous study, 84 monoclonal antibodies (mAbs) against hemagglutinin (HA) were prepared. When identifying their characteristics, it was revealed that the H1-84mAb not only binds to the HA antigen, but also cross-reacts with human brain tissue, suggesting that H1N1 influenza virus HA and human brain tissue have a heterophilic antigen (8). Heterophilic antigens are a class of common antigens that are unrelated between species, and exist in humans, animals and microorganisms (9).…”

Antibodies against H1N1 influenza virus cross‐react with α‐cells of pancreatic islets

“…Animal experiments were approved by the Institutional Animal Care and Use Committee of Shaanxi Provincial People's Hospital (Shaanxi, Xi'an). The preparation of mAbs has been reported previously 7 .…”

“…The authors proposed that these cross-reactive antibodies are associated with immune complex-mediated disease after H1N1 infection. 6 To investigate the possible effect of antibodies against H1N1 influenza virus, we used the type A H1N1 virus vaccine as an antigen and generated 84 mouse monoclonal antibodies (mAbs), and found that some mAbs could cross-react with human tissues 7 . In the present study, we further explored the topography of the immunoreactivity of the antibodies in pancreatic tissues.…”