两株H1N1亚型猪流感病毒HA蛋白中和性单克隆抗体的制备及抗病毒活性研究

doi:10.11843/j.issn.0366-6964.2018.07.015

Abstract

Abstract:

This study aimed to prepare monoclonal antibodies(MAbs) against HA protein of Eurasian avian-like H1N1(EA H1N1) virus and analyze their biological activities. After viral proliferation in chicken embryos, the purified whole-virus protein of A/swine/Henan/11/2005(HN11) was used to immunize BALB/c mice. The splenic cells of the mice were fused with the SP2/0 cells after the third immunization. After screening by indirect ELISA and hemagglutination inhibition test and three circles of sub-clone, two hybridoma cell strains secreting MAbs against HA protein of the HN11 were obtained, which was designated as 2B6 and 4C7, respectively. The ELISA titers of two MAbs were higher than 1:10⁷, and HI titer were higher than 5 log2. These two MAbs could specifically recognize HA protein expressed by plasmid pCAGGS-H1HA in 293T cells by IFA, but not in Western-blot assay. The results of neutralization test indicated that two MAbs have specific neutralization activities for EA H1N1. The protective efficacies of two MAbs were further verified in BALB/c mice, which suggested that MAbs 2B6 and 4C7 can effectively provide protection against the infection with homologous and heterologous H1N1 influenza viruses. Two MAbs, with high viral neutralization activity and strong anti-viral infection ability, were prepared, could be used for further research on anti-viral infection and the molecular basis of antigenic variation of EA H1N1 SIVs.

CLC Number:

S852.659

LIANG Wen-hua, JIA Yun-hui, XU Cheng-zhi, WU Yun-pu, CHEN Yan, YANG Huan-liang, QIAO Chuan-ling, CHEN Hua-lan. Preparation and Antiviral Activity of Two Neutralizing Monoclonal Antibodies against HA Protein of H1N1 Swine Influenza Virus[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(7): 1460-1466.

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References

[1] BAUDON E, PEYRE M, PEIRIS M, et al. Epidemiological features of influenza circulation in swine populations:A systematic review and meta-analysis[J]. PLoS One, 2017, 12(6):e0179044.
[2] PENSAERT M, OTTIS K, VANDEPUTTE J, et al. Evidence for the natural transmission of influenza A virus from wild ducts to swine and its potential importance for man[J]. Bull World Health Organ, 1981, 59(1):75-78.
[3] LIU J H, BI Y H, QIN K, et al. Emergence of European avian influenza virus-like H1N1 swine influenza A viruses in China[J]. J Clin Microbiol, 2009, 47(8):2643-2646.
[4] VINCENT A, AWADA L, BROWN I, et al. Review of influenza A virus in swine worldwide:A call for increased surveillance and research[J]. Zoonoses Public Health, 2014, 61(1):4-17.
[5] YANG H L, QIAO C L, TANG X, et al. Human infection from avian-like influenza A (H1N1) viruses in pigs, China[J]. Emerg Infect Dis, 2012, 18(7):1144-1146.
[6] ROVIDA F, PIRALLA A, MARZANI F C, et al. Swine influenza A (H1N1) virus (SIV) infection requiring extracorporeal life support in an immunocompetent adult patient with indirect exposure to pigs, Italy, October 2016[J]. Euro Surveill, 2017, 22(5):30456.
[7] YANG H L, CHEN Y, QIAO C L, et al. Prevalence, genetics, and transmissibility in ferrets of Eurasian avian-like H1N1 swine influenza viruses[J]. Proc Natl Acad Sci U S A, 2016, 113(2):392-397.
[8] BYRD-LEOTIS L, CUMMINGS R D, STEINHAUER D A. The interplay between the host receptor and influenza virus hemagglutinin and neuraminidase[J]. Int J Mol Sci, 2017, 18(7):1541.
[9] SHVARTSMAN D E, KOTLER M, TALL R D, et al. Differently anchored influenza hemagglutinin mutants display distinct interaction dynamics with mutual rafts[J]. J Cell Biol, 2003, 163(4):879-888.
[10] SKEHEL J J, WILEY D C. Receptor binding and membrane fusion in virus entry:The influenza hemagglutinin[J]. Annu Rev Biochem, 2000, 69:531-569.
[11] CHEN Y X, QIN K, WU W L, et al. Broad cross-protection against H5N1 avian influenza virus infection by means of monoclonal antibodies that map to conserved viral epitopes[J]. J Infect Dis, 2009, 199(1):49-58.
[12] NAUNDORF S, PREITHNER S, MAYER P, et al. In vitro and in vivo activity of MT201, a fully human monoclonal antibody for pancarcinoma treatment[J]. Int J Cancer, 2002, 100(1):101-110.
[13] BHAT S, BHATIA S, SOOD R, et al. Production and characterization of monoclonal antibodies against nucleoprotein of avian influenza virus[J]. Monoclon Antib Immunodiagn Immunother, 2013, 32(6):413-418.
[14] CHEN L M, RIVAILLER P, HOSSAIN J, et al. Receptor specificity of subtype H1 influenza A viruses isolated from swine and humans in the United States[J]. Virology, 2011, 412(2):401-410.
[15] OKUNO Y, ISEGAWA Y, SASAO F, et al. A common neutralizing epitope conserved between the hemagglutinins of influenza A virus H1 and H2 strains[J]. J Virol, 1993, 67(5):2552-2558.
[16] EKIERT D C, WILSON I A. Broadly neutralizing antibodies against influenza virus and prospects for universal therapies[J]. Curr Opin Virol, 2012, 2(2):134-141.

Preparation and Antiviral Activity of Two Neutralizing Monoclonal Antibodies against HA Protein of H1N1 Swine Influenza Virus

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