HIV毒株分离及体外感染实验研究

Isolation of HIV strain and in vitro virus infection experiments

目的 从艾滋病病毒 (HIV)感染者 (万某 )分离获得HIV毒株 ,应用此毒株在体外攻击与HIV感染者长期无保护性行为未引发感染者 (张某 )及健康人的淋巴细胞 ,检测病毒感染细胞情况 ,研究抗感染保护的机制。方法 采用RT测定法和P2 4抗原检测法。将HIV感染者淋巴细胞与健康人淋巴细胞共培养 ,并将获得的HIV毒株体外攻击张某及健康人的淋巴细胞 ,分别观察细胞病变并检测P2 4抗原及逆转录酶的活性。结果 从万某血中分离获得了HIV 1毒株 ,连续 8周显微镜观察 ,从第 2周开始可见HIV特有的细胞病变 ,逆转录酶的活性检测和P2 4抗原检测结果从第 2周为阳性 ,说明病毒分离成功。健康人淋巴细胞自HIV 1感染后 2周开始 ,显微镜可观察到HIV 1特有的细胞病变 ,逆转录酶的活性检测和P2 4抗原检测结果均为阳性 ,表明HIV侵入。张某的淋巴细胞在HIV感染后显微镜下观察 ,始终未见特有的细胞病变 ,逆转录酶的活性检测和P2 4抗原检测结果均为阴性 ,表明HIV病毒未侵入细胞。结论 分离的毒株可以感染健康人淋巴细胞 ,说明病毒本身无变异 ;病毒不能侵入张某的淋巴细胞 ,可能是其淋巴细胞表面HIV 1受体或辅受体的基因或功能发生突变 ,导致病毒无法侵入机体。

Objective A HIV strain was isolated from a HIV carrier(Mr.Wang)and used to attack lymphocytes collected from a healthy person and his sex partner(Ms.Zhang)who was free from HIV despite long time unprotected sex between them so as to study cell infections and anti-infection mechanism.Methods Lymphocytes from the HIV carrier and the healthy person were co-cultivated and tested by reverse transcriptase assay(RT)and HIV P24 antigen detection.The HIV strain was used to attack the lymphocytes collected from this healthy person and also from Ms.Zhang.ResultsAfter two-week cultivation,lymphocytes from Wang showed positive RT and P24 Ag activities and also pathomor phological changes.Lymphocytes from Zhang attacked by HIV strain were not infected while those from the healthy person were infected.Conclusion The anti-HIV phenomenon is probably associated with gene or functional mutation of HIV-1 receptors or coreceptors on the lymphocyte surface.