Identification and characterization of epitopes from influenza A virus hemagglutinin that induce broadly cross-reactive antibodies

Li,Yuan; Hu,Hanyu; Qi,Zongli; Sun,Jingying; Li,Yan; Feng,Qing; Guo,Chunyan; Wang,Haifang; Zhao,Penghua; Liu,Yang; Zhao,Xiangrong; Wang,Guanghua; Zhang,Hai; Liu,Libin; Hu,Jun

doi:10.3892/ijmm.2017.3344

Identification and characterization of epitopes from influenza A virus hemagglutinin that induce broadly cross-reactive antibodies

Authors:
- Yuan Li
- Hanyu Hu
- Zongli Qi
- Jingying Sun
- Yan Li
- Qing Feng
- Chunyan Guo
- Haifang Wang
- Penghua Zhao
- Yang Liu
- Xiangrong Zhao
- Guanghua Wang
- Hai Zhang
- Libin Liu
- Jun Hu
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Affiliations: Central Experimental Laboratory, Shaanxi Provincial People's Hospital, Key Laboratory of Infection and Immunity of Shaanxi Province, Xi'an, Shaanxi 710068, P.R. China, School of Public Health, Sun Yat‑sen University, Guangzhou, Guangdong 510080, P.R. China, Center of Experimental Animals, The 4th Military Medical University, Xi'an, Shaanxi 710032, P.R. China, Department of Pharmacy Medical College of Xi'an Peihua University, Xi'an, Shaanxi 710125, P.R. China
Published online on: December 22, 2017 https://doi.org/10.3892/ijmm.2017.3344
Pages: 1673-1682

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Abstract

Influenza is the most common infectious disease and is caused by influenza A virus (IAV) infection. Hemagglutinin (HA) is an important viral protein of influenza A and is a major component of current IAV vaccines. The side effects associated with IAV vaccination are well studied; however, the HA‑induced immunopathological changes have remained largely elusive. The primary objective of the present study was to determine the tissue cross‑reactive epitopes of HA proteins. Monoclonal antibodies (McAbs) were generated according to traditional methods using purified HA proteins from influenza vaccine lysates. The specificity of these McAbs was analyzed using western blot analysis and ELISA. Human tissue microarrays were employed for immunohistochemical staining to screen these McAbs. Rat brain tissues were subjected to immunohistochemical staining and electron microscopy to demonstrate the subcellular localization of antibodies targeting specific antigens. A total of 67 hybridoma cell lines positive for McAb against HA antigen were obtained. Three cross‑reactive McAbs (H1‑13, H1‑15 and A1‑10) were discovered through tissue screening. Based on the 3 cross‑reactive McAbs and the amino acid sequence of HA, the presence of two broadly cross‑reactive HA epitopes, 194‑WGIHH‑198 and 365‑WYGYHH‑370, was assumed. McAbs against these synthetic epitope peptides were obtained. They reacted with porphyrin ring‑containing molecules, including hemoglobin (Hb) and protoporphyrin, and with numerous types of normal tissue. In conclusion, the present study identified two broadly cross‑reactive epitopes on HA (194‑WGIHH‑198 and 365‑WYGYHH‑370). Antibodies against these epitopes react with Hb and numerous types of important normal tissues/organs. These newly identified cross‑reactive epitopes from IAV HA may provide crucial information for influenza research.

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