应用ELISPOT方法筛选确定尼帕病毒融合糖蛋白和附着糖蛋白的H-2 d限制性T细胞表位  

Screening and identification of H-2d-restricted T cell epitopes in fusion and attachment glycoproteins of Nipah virus by ELISPOT

作　　者：黄梦婧 邓瑶 赵志敏 陈晋妮 任皎[2] 王文[2] 沈晓玲 谭文杰[2] Huang Mengjing;Deng Yao;Zhao Zhimin;Chen Jinni;Ren Jiao;Wang Wen;Shen Xiaoling;Tan Wenjie(Department of Microbiology,Basic Medical College,Inner Mongolia Medical University,Hohhot 010010,China;National Health Commission Key Laboratory of Biosafety,National Institute for Viral Disease Control and Prevention,Chinese Center for Disease Control and Prevention,Beijing 102206,China;School of Public Health,Xinxiang Medical College,Xinxiang 453000,China)

机构地区：[1]内蒙古医科大学基础医学院微生物教研室,呼和浩特010010 [2]中国疾病预防控制中心病毒病预防控制所,国家卫生健康委员会生物安全重点实验室,北京102206 [3]新乡医学院公共卫生学院,新乡453000

出　　处：《中华微生物学和免疫学杂志》2022年第4期287-292,共6页Chinese Journal of Microbiology and Immunology

基　　金：国家重点研发计划(2016YFD0500300)。

摘　　要：目的:在小鼠中筛选和确定尼帕病毒(Nipah virus,NiV)融合糖蛋白(fusion glycoprotein,F)和附着糖蛋白(attachment glycoprotein,G)特异性的H-2 d限制性T细胞表位。 方法:本研究基于NiV F和G蛋白全序列设计合成多肽库(单肽长为15个氨基酸,前后肽重复10个氨基酸),使用表达NiV F和G蛋白的DNA疫苗肌肉注射免疫BALB/c小鼠,通过矩阵设计将F和G抗原全序列肽库分别混合成肽池,取免疫后小鼠脾细胞进行IFN-γ ELISPOT检测,确定肽库中特异反应的优势表位肽。结果:F肽库筛选到12条特异性优势肽,第56条多肽产生的反应最强;G肽库筛选到4条特异性优势肽,第72条多肽产生的反应最强。结论:本研究使用IFN-γ ELISPOT方法筛选和确定了12条NiV F抗原特异性和4条NiV G抗原特异性H-2 d限制性优势T细胞表位,为NiV免疫学与疫苗研究提供了参考。Objective To screen and identify H-2d-restricted T cell epitopes in fusion(F)and attachment(G)glycoproteins of Nipah virus(NiV)in mice.Methods The complete peptides(single peptide contains 15 amino acids,and 10 amino acids were repeated in the front and back peptides)derived from F and G antigens were mixed into peptide libraries.BALB/c mice were immunized with DNA vaccines expressing NiV F and G proteins alone and in combination.The full sequence peptide libraries of F and G antigens were mixed into peptide pools by matrix design,and spleen cells of immunized mice were collected and analyzed by IFN-γELISPOT assay to detect the dominant H-2d-restricted epitope peptides.Results Twelve dominant H-2d-restricted peptides were screened from the F protein-specific peptide library and the 56th peptide produced the strongest reaction.Four dominant peptides were screened from the G protein-specific peptide library and the 72nd peptide produced the strongest reaction.Conclusions In this study,12 F antigen-specific and 4 G antigen-specific H-2d restricted dominant T cell epitopes of NiV were screened and identified by IFN-γELISPOT,which could provide reference for immunological analysis of NiV and vaccine research.

关 键 词：尼帕病毒 H-2 d限制 T细胞表位 IFN-γELISPOT 

分 类 号：R392[医药卫生—免疫学]