HIV感染人CD4+T细胞生物学特性的实验研究

目的观察人免疫缺陷病毒（HIV）-1SF33感染后MT4细胞超微结构、细胞周期、细胞内钙离子浓度改变，为寻找艾滋病发病机制提供研究基础。方法以HIV-1SF33实验株感染MT4细胞，72h后用P24试剂盒双抗体夹心法检测HIV抗原表达情况以验证感染是否成功。设正常MT4细胞组与HIV感染MT4细胞组，用倒置显微镜观察细胞形态，透射电镜观察细胞超微结构，激光共聚焦显微镜检测细胞内钙离子浓度，流式细胞仪检测细胞周期。结果染毒成功。染毒组细胞多散在排列，可见多核巨细胞和空泡状细胞形成；胞浆内可见大量大小均一的HIV颗粒堆积，细胞器肿胀，内质网、线粒体扩张，溶酶体体积增大，数量增多，部分细胞融合，胞浆内形成多个空泡；细胞内钙离子含量增高；HIV感染与未感染MT4细胞相比，G2-M-S期细胞比例减少，G0-G1期细胞比例增多。结论HIV感染CD4^＋T细胞后，可直接导致CD4^＋T细胞病变，如细胞肿胀，细胞内钙离子含量增高，细胞增殖减慢。

The biological study of HIV infected human CD4+T cells

Objective To study the pathogenesis of AIDS by observing the ultrastructure,cell cycle,intracellular calcium concentration of MT4 cells after being infected by HIV-1_(SF33).Methods Human MT4 cells(CD4~ ) were infected by HIV strain SF33.The expression of HIV p24 antigen was tested by sandwich enzyme immunoassay.Two groups(the group of MT4 cells and HIV-1 infected MT4 cells) were divided.The cell morphology was observed by inverted microscope,the cell ultrastructure was observed by transmission electronmicroscope,the intracellular Ca~(2 ) concentration was measured by laser confocal scanning microscope after dying with flue-3 AM,the cell marker was analysed by flow cytometry.Results The cell cycle infection was successfully established after HIVp24 antigen was detected.Multinucleated giant cell and vacuolar cell could be seen in HIV infected MT4 cells.The cells displayed swelling,the dilation of mitochondria and other organelles could be seen,the lysosome became more and larger after infection.The Ca~(2 ) concentration of HIV infected MT4 cells were higher than that of MT4 cells. The proportion of HIV infected cells in G2-M-S phase was lower than MT4 cells.Conclusion HIV infection can result in a series of pathological changes,including cell proliferation,cell morphology and intracellular calcium ions etc.