日本乙型脑炎病毒强、弱毒株非结构蛋白NS1、NS2A和NS1-NS2A诱导小鼠CD4+T细胞应答的研究

doi:10.11843/j.issn.0366-6964.2018.08.014

Abstract

Abstract:

In order to understand the immunological effect of nonstructural protein of Japanese encephalitis virus (JEV), we analyzed the eukaryotic expression differences of NS1, NS2A and NS1-2A, nonstructural proteins of JEV. CPE was observed on BHK-21 cells by observing JE live attenuated vaccine (SA14-14-2 strain) and JEV Guizhou isolate (GZ strain) respectively. Cell suspensions of virulent, attenuated strains were collected and their total RNA were extracted. The NS1, NS2A and NS1-2A genes were amplified by RT-PCR, and six eukaryotic expression plasmids, pcDNA3.1 (+)-NS1r, pcDNA3.1 (+)-NS2Ar, pcDNA3.1 (+)-NS1-2Ar, pcDNA3.1 (+)-NS1q, pcDNA3.1 (+)-NS2Aq and pcDNA3.1 (+)-NS1-2Aq, were constructed successfully. The healthy mice were immunized with the above plasmids, and the blood and tissues of those mice were collected for JEV antibody detection and cell immunoassay respectively. The results showed that JEV GZ virulent strain and JEV SA14-14-2 virulent strain could proliferate and produce CPE on BHK21 cells; The results of routine blood tests and T lymphocyte subsets detection showed that the 6 eukaryotic expression plasmids immunization in mice increased the levels of CD4⁺ and CD8⁺ cells significantly or extremely significantly. The results indicated that the cellular immunity produced by the non-structural proteins of the attenuated strain JEV was slightly better than that produced by the non-structural proteins of the attenuated strains, but the difference was not significant. These results provide reference for the basic research of encephalitis virus pathogens.

CLC Number:

S852.659.6

YANG Wei, WEI Guan-dong, TANG De-yuan, ZENG Zhi-yong, HUANG Tao, WANG Bin, HU Ling-ling, LONG Dong-mei, HUANG Qiu-han, LIAO Xiao-kang, TIAN Hong-yu. Japanese Encephalitis Virus Virulent Strains and Attenuated Strains of Non-Structural Proteins NS1, NS2A and NS1-NS2A Induce Mouse CD4⁺ T Cell Responses[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(8): 1693-1700.

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Japanese Encephalitis Virus Virulent Strains and Attenuated Strains of Non-Structural Proteins NS1, NS2A and NS1-NS2A Induce Mouse CD4⁺ T Cell Responses

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Japanese Encephalitis Virus Virulent Strains and Attenuated Strains of Non-Structural Proteins NS1, NS2A and NS1-NS2A Induce Mouse CD4+ T Cell Responses

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Japanese Encephalitis Virus Virulent Strains and Attenuated Strains of Non-Structural Proteins NS1, NS2A and NS1-NS2A Induce Mouse CD4⁺ T Cell Responses