Virologica Sinica

, Volume 34, Issue 3, pp 306–314 | Cite as

Localization Analysis of Heterophilic Antigen Epitopes of H1N1 Influenza Virus Hemagglutinin

  • Chun-Yan Guo
  • Hai-Xiang Zhang
  • Jun-Jun Zhang
  • Li-Jun Sun
  • Hui-Jin Li
  • Dao-Yan Liang
  • Qing Feng
  • Yan Li
  • Yang-Meng Feng
  • Xin XieEmail author
  • Jun HuEmail author
Research Article


Previous studies have indicated that two monoclonal antibodies (mAbs; A1-10 and H1-84) of the hemagglutinin (HA) antigen on the H1N1 influenza virus cross-react with human brain tissue. It has been proposed that there are heterophilic epitopes between the HA protein and human brain tissue (Guo et al. in Immunobiology 220:941–946, 2015). However, characterisation of the two mAbs recognising the heterophilic epitope on HA has not yet been performed. In the present study, the common antigens of influenza virus HA were confirmed using indirect enzyme-linked immunosorbent assays and analysed with DNAMAN software. The epitopes were localized to nine peptides in the influenza virus HA sequence and the distribution of the peptides in the three-dimensional structure of HA was determined using PyMOL software. Key amino acids and variable sequences of the antibodies were identified using abYsis software. The results demonstrated that there were a number of common antigens among the five influenza viruses studied that were recognised by the mAbs. One of the peptides, P2 (LVLWGIHHP191–199), bound both of the mAbs and was located in the head region of HA. The key amino acids of this epitope and the variable regions in the heavy and light chain sequences of the mAbs that recognised the epitope are described. A heterophilic epitope on H1N1 influenza virus HA was also introduced. The existence of this epitope provides a novel perspective for the occurrence of nervous system diseases that could be caused by influenza virus infection, which might aid in influenza prevention and control.


H1N1 influenza virus HA antigen Monoclonal antibody Localization Heterophilic epitope 



This work was supported by The National Key Research and Development Program of China (Grant No. 2016YFD0500700), The Natural Science Basic Research Program of Shaanxi Province (Grant No. 2016JM8065) and Shaanxi Provincial People’s Hospital Incubation Fund Program (Grant No. 2015YX-4).

Author Contributions

CG, XX and LS designed the experiments. CG, JZ, HL, DL, QF, YL and YF conducted the experiments. CG and JH analyzed the data. CG and HZ wrote the paper. All authors approved the final manuscript.

Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no competing interests.

Animal and Human Rights Statement

This article does not include any experiments that involve human or animal subjects.

Supplementary material

12250_2019_100_MOESM1_ESM.pdf (316 kb)
Supplementary material 1 (PDF 55 kb)


  1. Ahmed SS, Volkmuth W, Duca J, Corti L, Pallaoro M, Pezzicoli A, Karle A, Rigat F, Rappuoli R, Narasimhan V, Julkunen I, Vuorela A, Vaarala O, Nohynek H, Pasini FL, Montomoli E, Trombetta C, Adams CM, Rothbard J, Steinman L (2015) Antibodies to influenza nucleoprotein cross-react with human hypocretin receptor 2. Sci Transl Med 7:294ra105CrossRefGoogle Scholar
  2. Blackmore S, Hernandez J, Juda M, Ryder E, Freund GG, Johnson RW, Steelman AJ (2017) Influenza infection triggers disease in a genetic model of experimental autoimmune encephalomyelitis. Proc Natl Acad Sci USA 114:E6107–E6116CrossRefGoogle Scholar
  3. Cong YJ, Ren FZ, Yun ZY (2010) Identification of Ig E-binding β-epitopes on β-lactoglobulin. Food Sci 15:190–193 (in Chinese) Google Scholar
  4. Coppieters KT, von Herrath M (2013) Antibody cross-reactivity and the viral aetiology of type 1 diabetes. J Pathol 230:1–3CrossRefGoogle Scholar
  5. de Wit E, Siegers JY, Cronin JM, Weatherman S, van den Brand JM, Leijten LM, van Run P, Begeman L, van den Ham HJ, Andeweg AC, Bushmaker T, Scott DP, Saturday G, Munster VJ, Feldmann H, van Riel D (2018) 1918 H1N1 influenza virus replicates and induces proinflammatory cytokine responses in extrarespiratory tissues of ferrets. J Infect Dis 217:1237–1246CrossRefGoogle Scholar
  6. Desdouits M, Munier S, Prevost MC, Jeannin P, Butler-Browne G, Ozden S, Gessain A, Van Der Werf S, Naffakh N, Ceccaldi PE (2013) Productive infection of human skeletal muscle cells by pandemic and seasonal influenza A(H1N1) viruses. PLoS ONE 8:e79628CrossRefGoogle Scholar
  7. Engmark M, Andersen MR, Laustsen AH, Patel J, Sullivan E, de Masi F, Hansen CS, Kringelum JV, Lomonte B, Gutiérrez JM, Lund O (2016) High-throughput immuno-profiling of mamba (Dendroaspis) venom toxin epitopes using high-density peptide microarrays. Sci Rep 6:36629CrossRefGoogle Scholar
  8. Forssman J (1911) Die Herstellung hochwertiger spezifischer Schafha¨monysine ohne Verwendung von Schafblut. Ein Beitrag zur Lehre von heterologer Antiko¨rpebildung. Biochem Z 37:78–115 (in German) Google Scholar
  9. Gong X, Yin H, Shi YH, Guan SS, He XQ, Yang L, Yu YJ, Kuai ZY, Jiang CL, Kong W, Wang S, Shan YM (2016) Conserved stem fragment from H3 influenza hemagglutinin elicits cross-clade neutralizing antibodies through stalk-targeted blocking of conformational change during membrane fusion. Immunol Lett 172:11–20CrossRefGoogle Scholar
  10. Guedj R, Desguerre I, Brassier A, Boddaert N, Hubert P, Oualha M (2012) Unusual muscular injury in an infant with severe H1N1 infection. Pediatr Neurol 47:51–54CrossRefGoogle Scholar
  11. Guo CY, Tang YG, Qi ZL, Liu Y, Zhao XR, Huo XP, Li Y, Feng Q, Zhao PH, Wang X, Li Y, Wang HF, Hu J, Zhang XJ (2015) Development and characterization of a panel of cross-reactive monoclonal antibodies generated using H1N1 influenza virus. Immunobiology 220:941–946CrossRefGoogle Scholar
  12. Hendrickson CM, Matthay MA (2013) Viral pathogens and acute lung injury: investigations inspired by the SARS epidemic and the 2009 H1N1 Influenza pandemic. Semin Respir Crit Care Med 34:475–486CrossRefGoogle Scholar
  13. Hu B, Wei YQ (2004) Molecular mimicry associated with infection, autoimmunity, and tumor immunotherapy. Chin Bull Life Sci 16:66–72 (in Chinese) Google Scholar
  14. Hurwitz ES, Nelson DB, Davis C, Morens D, Schonberger LB (1982) National surveillance for Reye syndrome: a five-year review. Pediatrics 70:895–900Google Scholar
  15. Ishida Y, Kawashima H, Morichi S, Yamanaka G, Okumura A, Nakagawa S, Morishima T (2015) Brain magnetic resonance imaging in acute phase of pandemic influenza A (H1N1)2009-associated encephalopathy in children. Neuropediatrics 46:20–25Google Scholar
  16. Khurana S, Verma S, Verma N, Crevar CJ, Carter DM, Manischewitz J, King LR, Ross TM, Golding H (2010) Properly folded bacterially expressed H1N1 hemagglutinin globular head and ectodomain vaccines protect ferrets against H1N1 pandemic influenza virus. PLoS ONE 5:e11548CrossRefGoogle Scholar
  17. Levin MC, Lee SM, Kalume F, Morcos Y, Dohan FC, Hasty K, Callaway JC, Zunt J, Desiderio D, Stuart JM (2002) Autoimmunity due to molecular mimicry as a cause of neurological disease. Nat Med 8:509–513CrossRefGoogle Scholar
  18. Li Y, Hu HY, Qi ZL, Sun JY, Li Y, Feng Q, Guo CY, Wang HF, Zhao PH, Liu Y, Zhao XR, Wang GH, Zhang H, Liu L, Hu J (2018) Identification and characterization of epitopes from influenza A virus hemagglutinin that induce broadly cross-reactive antibodies. Int J Mol Med 41:1673–1682Google Scholar
  19. Mammas IN, Koutsaftiki C, Papantzimas K, Symeonoglou Z, Koussouri M, Theodoridou M, Myriokefalitakis N (2011) Thrombocytic thrombocytopenic purpura in a child with A/H1N1 influenza infection. J Clin Virol 51:146–147CrossRefGoogle Scholar
  20. Marchalonis JJ, Adelman MK, Robey IF, Schluter SF, Edmundson AB (2001) Exquisite specificity and peptide epitope recognition promiscuity, properties shared by antibodies from sharks to humans. J Mol Recognit 14:110–121CrossRefGoogle Scholar
  21. Nair DT, Singh K, Sahu N, Rao KV, Salunke DM (2000) Crystal structure of an antibody bound to an immunodominant peptide epitope: novel features in peptide-antibody recognition. J Immunol 165:6949–6955CrossRefGoogle Scholar
  22. Nair DT, Singh K, Siddiqui Z, Nayak BP, Rao KV, Salunke DM (2002) Epitope recognition by diverse antibodies suggests conformational convergence in an antibody response. J Immunol 168:2371–2382CrossRefGoogle Scholar
  23. Robey IF, Edmundson AB, Schluter SF, Yocum DE, Marchalonis JJ (2002) Specificity mapping of human anti-T cell receptor monoclonal natural antibodies: defining the properties of epitope recognition promiscuity. FASEB J 16:642–652CrossRefGoogle Scholar
  24. Sarkanen TO, Alakuijala APE, Dauvilliers YA, Partinen MM (2018) Incidence of narcolepsy after H1N1 influenza and vaccinations: systematic review and meta-analysis. Sleep Med Rev 38:177–186CrossRefGoogle Scholar
  25. Seow J, Morales RA, MacRaild CA, Krishnarjuna B, McGowan S, Dingjan T, Jaipuria G, Rouet R, Wilde KL, Atreya HS, Richards JS, Anders RF, Christ D, Drinkwater N, Norton RS (2017) Structure and characterisation of a key epitope in the conserved C-terminal domain of the malaria vaccine candidate MSP2. J Mol Biol 429:836–846CrossRefGoogle Scholar
  26. Srinivasappa J, Saegusa J, Prabhakar BS, Gentry MK, Buchmeier MJ, Wiktor TJ, Koprowski H, Oldstone MB, Notkins AL (1986) Molecular mimicry: frequency of reactivity of monoclonal antiviral antibodies with normal tissues. J Virol 57:397–401Google Scholar
  27. Taylor DN, Treanor JJ, Strout C, Johnson C, Fitzgerald T, Kavita U, Ozer K, Tussey L, Shaw A (2011) Induction of a potent immune response in the elderly using the TLR-5 agonist, flagellin, with a recombinant hemagglutinin influenza-flagellin fusion vaccine (VAX125, STF2. HA1 SI). Vaccine 29:4897–4902CrossRefGoogle Scholar
  28. Vellozzi C, Iqbal S, Broder K (2014) Guillain-Barre syndrome, influenza, and influenza vaccination: the epidemiologic evidence. Clin Infect Dis 58:1149–1155CrossRefGoogle Scholar
  29. Wang JJ, Yang GX, Zhang WC, Lu L, Tsuneyama K, Kronenberg M, Véla JL, Lopez-Hoyos M, He XS, Ridgway WM, Leung PS, Gershwin ME (2014) Escherichia coli infection induces autoimmune cholangitis and anti-mitochondrialantibodies in non-obese diabetic (NOD).B6 (Idd10/Idd18) mice. Clin Exp Immunol 175:192–201CrossRefGoogle Scholar
  30. Wilking AN, Elliott E, Garcia MN, Murray KO, Munoz FM (2009) Central nervous system manifestations in pediatric patients with influenza A H1N1 infection during the 2009 pandemic. Pediatr Neurol 51:370–376CrossRefGoogle Scholar
  31. Xu W, Han L, Lin Z (2011) Screening of random peptide library of hemagglutinin from pandemic 2009 A (H1N1) influenza virus reveals unexpected antigenically important regions. PLoS ONE 6:e18016CrossRefGoogle Scholar

Copyright information

© Wuhan Institute of Virology, CAS 2019

Authors and Affiliations

  1. 1.Shaanxi Province Research Center of Cell Immunological Engineering and TechnologyCentral Laboratory of Shaanxi Provincial People’s HospitalXi’anChina
  2. 2.Xian Yang Municipal Center for Disease Control and PreventionXianyangChina
  3. 3.Shaanxi Key Laboratory of Ischemic Cardiovascular Disease, Institute of Basic and Translational MedicineXi’an Medical UniversityXi’anChina
  4. 4.Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, College of Life SciencesNorthwest UniversityXi’anChina

Personalised recommendations