表达新疆出血热病毒S基因片段的重组腺病毒特异性CTL功能测定

目的通过表达新疆出血热病毒(XHFV)S基因片段的重组腺病毒特异性细胞毒性T淋巴细胞(CTL)功能测定,探求由XHFVS基因片段编码的核衣壳蛋白作为抗原诱导的特异性CTL细胞对重组腺病毒致敏的靶细胞杀伤能力大小。方法PCR扩增XHFVS基因的cDNA片段,将扩增后的cDNA片段克隆入腺病毒自主载体,使其成为重组腺病毒,用重组腺病毒感染Balb/c小鼠后,利用非放射性乳酸脱氢酶(LDH)释放法测定重组腺病毒的特异性CTL功能。结果在一定比例范围内,效应细胞与靶细胞的比例越高,特异性CTL%越高。结论由XHFVS基因片段编码的核衣壳蛋白抗原诱导的特异性CTL细胞,能特异性地杀伤重组腺病毒致敏的靶细胞;证实由腺病毒自主载体本身所表达的一些蛋白质也可诱导细胞免疫,由该载体所引发的细胞免疫对疫苗接种不利。

The assay of specific CTL function of recombinant adenovirus expressing HFV S gene fragment

Objective To explore the cytotoxic ability of specific CTL cells induced by the nucleocpsid protein antigen encoded by the S gene fragment of XHFV to the target cells sensitized by recombinant adenovirus through the assay of special CTL function of recombinant adenovirus expressing XHFV S gene fragment.Methods S genetic cDNA fragment amplified by PCR was cloned into the helper-free vector of adenovirus,which made the helper-free vector of adenovirus the recombinant adenovirus. After they were used to infect the Balb/c mice,specific CTL function of recombinant adenovirus was detected with the non-radioactive LDH release assay.Results In the suitable ratio of effector cells to target cells, the higher the ratio of effector cells to target cells is,the higher specific CTL percentage is.Conclusions Specific CTL function induced by the nucleocapsid protein antigen encoded by S gene fragment of XHFV can cause special damage to the CTL bears out that some protein expressed by the helper-free vector of adenovirus can induce cellular immunity,which is deteriorative to vaccine inoculation with the recombinant adenovirus.