重组人穿孔素协同TNF-α对HIV感染CD4^＋细胞的影响

目的观察重组人穿孔素协同TNF-α对HIV感染CD4^＋细胞的影响。方法HIV-1SF33感染MT4细胞，加入重组人穿孔素和TNF-α作用24h。倒置显微镜观察细胞形态，单细胞凝胶电泳技术观察DNA损伤情况，吖啶橙／溴乙啶双荧光染色和原位（缺口）末端标记法检测细胞凋亡。结果HIV感染MT4细胞形态尚好，仅出现荧光团，可见个别凋亡细胞；单纯TNF-α作用后，胞浆内可见少量颗粒，部分细胞核出现弥散和微量的彗星拖尾现象，凋亡细胞较正常对照组增多；穿孔素和TNF-α协同作用后，部分细胞体积变小，细胞内颗粒增多，明显的彗星拖尾现象，凋亡细胞数增多。结论穿孔素协同作用可明显增强TNF-α致HIV感染CD4^＋细胞的凋亡。

Effect of Recombinant Human Perforin and TNF-α on the HIV Infected CD4^＋ Cells

Objective To observe the synergic effect of the recombinant human perforin and tumor necrosis factor-α（TNF-α） on the HIV infected CD4^＋ ceils. Methods In the present study, the human CD4^＋ cells were infected by HIV strain SF33 successfully. TNF-α and recombinant human perforin was added to the HIV infected CD4^＋ cells. The ceil morphology was observed by inverted microscope. The single ceil gel electropherosis was used to observe DNA observe the ceil apoptosis. Results In the was seen, no obvious tailing damage, AO/EB double fluorescence and TUNEL were used to control group, the cell morphology was good; only fluorophore ; there were only individual apoptosis ceils seen. In TNF-α treated group, the clarity was decreased, a small quantity of particles were seen in the cytoplasm. The diffusion of cell nucleus and a few comets tailing of part cells were seen. more apoptosis ceils were increased than those in the control group. After the synergic effect of recombinant human perforin and TNF-α, the part of ceils shrinked, the particles were increased in the cytoplasm, the obvious comet tailings were seen. The apoptosis cells were increased significantly. Conclusion Recombinant human perforin can increase obviously the effect that TNF-α induced the apoptosis of HIV infected CD4^＋ ceils. The research gives an important guidance for how to utilize the synergetic function of immunological molecules to treat AIDS.