鸡传染性支气管炎病毒S1蛋白MHCⅠ分子限制性T细胞表位的鉴定

doi:10.11843/j.issn.0366-6964.2016.03.019

Abstract

Abstract:

A T cell epitope candidate peptide Sp6 of the avian infectious bronchitis virus S1 was predicted with biological structures technology.To identify this peptide exist in S1 protein and verify validity of Sp6，the recombinant plasmid pCAGGS-S1 was constructed.While it can be expressed in eukaryotic cells after determined by indirect immunofluorescence and Western blot，using the recombinant plasmid immune SPF chickens，and the antibodies against S1 were detected by ELISA.Splenic cells were collected from inoculated chickens and stimulated with the peptides Sp6，NP_89-97(PKKTGGPIY) and negative control.By SBRY Green qPCR detecting IFN-γ production of spleen lymphocytes and flow cytometry detecting the multiplication of CD8⁺ T lymphocytes，preliminarily make sure that the candidate peptide is correct.The IFA Western blot and ELISA results showed that the expression of S1 was confirmed in 293T and can induced the humoral immune，indicate that S1 protein is reaction to the original and lied a foundation for further validation of T cell epitope；the results of SBRY Green qPCR showed that spleen lymphocytes of chickens stimulated with the peptide Sp6 secreted significantly-increased IFN-γ.Flow cytometry assay results showed that the CD8⁺T lymphocytes were proliferated by 34.8%，2.6%，and 0，respectively，which indicated that the peptide Sp6 was a T cell epitope of IBV S1 and it can induce activation of chicken lymphocytes to produce cytotoxic T lymphocyte (CTL) reaction in vivo.All above results indicate that the peptide Sp6 was a T cell epitope of IBV S1 and it can induce activation of chicken lymphocytes to produce CTL reaction in vivo.The results possesses significance to mechanism of protective immunity against avian infectious bronchitis virus and the research towards universal IBV vaccine.

CLC Number:

S852.659.6

ZHU Feng-zhu，LU Mei，HUANG Qing-hua，YANG Shao-hua1，HUANG Yan-yan，WU Jia-qiang，TAN Liu-gang，ZHANG Xiu-mei，CUI Yan-shun，XU Chuan-tian . Identification of the MHC Class Ⅰ-restricted Cytotoxic T Cell Epitope in Avian Infectious Bronchitis Virus S1 Protein[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(3): 559-565.

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Identification of the MHC Class Ⅰ-restricted Cytotoxic T Cell Epitope in Avian Infectious Bronchitis Virus S1 Protein

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