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

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