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为探讨汉滩病毒核衣壳蛋白 (HTNV NP )诱生的特异性细胞免疫的分子基础 ,为阐明HFRS的发病机制和疫苗的设计提供资料 ,本文分离HFRS患者的PBMC ,建立HTNV NP特异性的CTL克隆 ,同时将克隆有HTNVS基因不同片段的真核表达载体转染患者自身的B淋巴母细胞系 (BLCL ) ,建立CTL靶细胞系 ,并进行细胞杀伤试验。建立的特异性CTL克隆对表达完整NP、NP羧基和氨基端肽段的靶细胞均有比较明显的杀伤效应 ,平均杀伤率分别为 5 0 2 %、 39 0 %和 2 5 8%。表明HTNV NP优势T细胞表位可能主要位于病毒核蛋白的羧基端
To investigate the molecular basis of specific cellular immunity induced by Hantaan virus nucleocapsid protein (HTNV NP) and to provide evidence for elucidating the pathogenesis of HFRS and the design of vaccines, we isolated PBMC from HFRS patients and established HTNV NP-specific CTL clones. At the same time, the eukaryotic expression vector cloned with different fragments of HTNVS gene was transfected into B lymphoblastoid cell line (BLCL) of patients. CTL target cell lines were established and cell killing assay was performed. The established specific CTL clone had obvious killing effect on target cells expressing intact NP, NP carboxyl and amino terminal peptide with average killing rates of 502, 39 0 and 25.8% respectively. It is indicated that the dominant T cell epitope of HTNV NP may be mainly located at the carboxyl terminus of the viral nucleoprotein