Prediction of common epitopes on hemagglutinin of the influenza A virus (H1 subtype)

Guo, Chunyan; Xie, Xin; Li, Huijin; Zhao, Penghua; Zhao, Xueqiang; Sun, Jingying; Wang, Haifang; Liu, Yang; Li, Yan; Hu, Qiaoxia; Hu, Jun; Li, Yuan

doi:10.1016/j.yexmp.2014.12.005

Cited by 7 publications

(10 citation statements)

References 19 publications

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“…McAbs produced by immunization of mice with IAV HA antigen. Sixty-seven hybridoma cell lines reactive to McAb against IAV HA antigen were obtained from 3 experiments; among these, 47 produced IgG class and 20 produced IgM class antibodies; 25 were H1 subtype-specific, and the remaining ones cross-reacted with the other HA subtypes, including 3 (H1-13, H1-15 and A1-10) that reacted not only with IAV HA but also with a wide variety of antigens as previously reported (31,32). The present study focused on these 3 cross-reactive McAbs.…”

Section: Resultsmentioning

confidence: 74%

“…Antibodies. Five McAbs (H1-13, A1-6, H1-15, A1-10 and H1-17) against the HA protein of H1N1 influenza virus were previously prepared in our laboratory (31).…”

Section: Mcab Preparationmentioning

confidence: 99%

“…McAb preparation. The feeder layer cells, immune spleen cells and myeloma Sp2/0 cells were routinely prepared, and the hybridoma were prepared using polyethylene glycol-mediated chemical fusion (31).…”

Section: Mcab Preparationmentioning

confidence: 99%

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

et al. 2017

Int J Mol Med

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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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Section: Resultsmentioning

confidence: 74%

“…Antibodies. Five McAbs (H1-13, A1-6, H1-15, A1-10 and H1-17) against the HA protein of H1N1 influenza virus were previously prepared in our laboratory (31).…”

Section: Mcab Preparationmentioning

confidence: 99%

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

et al. 2017

Int J Mol Med

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“…When preparing for fusion, the mice were injected intraperitoneally with influenza vaccine on the first day and began to fuse on the third day. Preparation of mAb: The process of cell fusion and the screening of positive clones were carried out according to the method of reference (Guo et al 2015 ). When the hybridoma cells reached 20–50% / well, the positive cells were screened by ELISA method.…”

Section: Methodsmentioning

confidence: 99%

Identification and analysis of B cell epitopes of hemagglutinin of H1N1 influenza virus

Qian

Huang²,

Feng

et al. 2022

Arch Microbiol

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The frequent variation of influenza virus hemagglutinin (HA) antigen is the main cause of influenza pandemic. Therefore, the study of B cell epitopes of HA is of great significance in the prevention and control of influenza virus. In this study, the split vaccine of 2009 H1N1 influenza virus was used as immunogen, and the monoclonal antibodies (mAbs) were prepared by conventional hybridoma fusion and screening techniques. The characteristics of mAbs were identified by ELISA method, Western-blot test and hemagglutination inhibition test (HI). Using the obtained mAbs as a tool, the B cell epitopes of HA were predicted by ELISA blocking test, sandwich ELISA method and computer simulation method. Finally, four mAbs against HA antigen of H1N1 influenza virus were obtained. The results of ELISA and computer prediction showed that there were at least two types of epitopes on HA of influenza virus. The results of this study complemented the existing methods for predicting HA epitopes, and also provided a new method for predicting other pathogenic microorganisms.

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“…Influenza has been for centuries a significant contributor to mortality and continues to be a significant threat to public health worldwide [ 1 , 2 ]. The influenza virus belongs to the Orthomyxoviridae family and is divided into four subtypes: A, B, C, and D [ 3 ].…”

Section: Introductionmentioning

confidence: 99%

In silico design of recombinant multi-epitope vaccine against influenza A virus

et al. 2022

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Background Influenza A virus is one of the leading causes of annual mortality. The emerging of novel escape variants of the influenza A virus is still a considerable challenge in the annual process of vaccine production. The evolution of vaccines ranks among the most critical successes in medicine and has eradicated numerous infectious diseases. Recently, multi-epitope vaccines, which are based on the selection of epitopes, have been increasingly investigated. Results This study utilized an immunoinformatic approach to design a recombinant multi-epitope vaccine based on a highly conserved epitope of hemagglutinin, neuraminidase, and membrane matrix proteins with fewer changes or mutate over time. The potential B cells, cytotoxic T lymphocytes (CTL), and CD4 T cell epitopes were identified. The recombinant multi-epitope vaccine was designed using specific linkers and a proper adjuvant. Moreover, some bioinformatics online servers and datasets were used to evaluate the immunogenicity and chemical properties of selected epitopes. In addition, Universal Immune System Simulator (UISS) in silico trial computational framework was run after influenza exposure and recombinant multi-epitope vaccine administration, showing a good immune response in terms of immunoglobulins of class G (IgG), T Helper 1 cells (TH1), epithelial cells (EP) and interferon gamma (IFN-g) levels. Furthermore, after a reverse translation (i.e., convertion of amino acid sequence to nucleotide one) and codon optimization phase, the optimized sequence was placed between the two EcoRV/MscI restriction sites in the PET32a+ vector. Conclusions The proposed “Recombinant multi-epitope vaccine” was predicted with unique and acceptable immunological properties. This recombinant multi-epitope vaccine can be successfully expressed in the prokaryotic system and accepted for immunogenicity studies against the influenza virus at the in silico level. The multi-epitope vaccine was then tested with the Universal Immune System Simulator (UISS) in silico trial platform. It revealed slight immune protection against the influenza virus, shedding the light that a multistep bioinformatics approach including molecular and cellular level is mandatory to avoid inappropriate vaccine efficacy predictions.

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Prediction of common epitopes on hemagglutinin of the influenza A virus (H1 subtype)

Cited by 7 publications

References 19 publications

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

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

Identification and analysis of B cell epitopes of hemagglutinin of H1N1 influenza virus

In silico design of recombinant multi-epitope vaccine against influenza A virus

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