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目的:探讨SIVmac239感染CD4+T淋巴细胞后,能否产生某些可抑制B淋巴细胞生长的活性因子,及其抑制作用的机制,为HIV/AIDS进一步的病原学研究和控制提供有益的线索。方法:用MTT实验观察不同时间收集的SIV-mac239感染的CD4+T淋巴细胞上清和未感染SIVmac239的CD4+T淋巴细胞的上清对猕猴B淋巴细胞生长的抑制作用,用3Hthymidine uptake assay判断这些上清是否抑制B淋巴细胞的DNA合成,用凝胶电泳方法检测DNA合成被抑制是否为凋亡的机制,用Western blot方法检测Cyclin D1的表达情况。结果:SIVmac239感染的CD4+T淋巴细胞上清含有抑制猕猴B淋巴细胞生长的因子,并且,其抑制效果随着作用时间的延长而增加。这些因子可较强的抑制CD4+T淋巴细胞的DNA合成,最高抑制率可达81%。凝胶试验结果未见DNA梯状条带。Western blot试验发现Cyclin D1表达被抑制。结论:SIVmac239感染CD4+T淋巴细胞可产生抑制B淋巴细胞生长的因子。此因子可较强的抑制猕猴B淋巴细胞DNA的合成,但并不引起B淋巴细胞凋亡。其主要机制为抑制猕猴B淋巴细胞的细胞周期蛋白D1的产生,使B淋巴细胞停止在G1期,即抑制作用的主要的机制为细胞被捕获在细胞周期的G1期。本研究结果发现一个新的现象,即猕猴B淋巴细胞虽然不被SIVmac239感染,但可被SIVmac239感染的CD4+T淋巴细胞产生的细胞因子所捕获,抑制其细胞DNA合成,从而可导致该细胞的功能发生障碍和影响机体体液免疫作用。
OBJECTIVE: To investigate whether SIVmac239 can infect CD4 + T lymphocytes to inhibit the growth of B lymphocytes and the mechanism of its inhibitory effect, and provide useful clues for the further etiological research and control of HIV / AIDS. Methods: MTT assay was used to observe the inhibitory effects of supernatant from CD4 + T lymphocytes infected with SIV-mac239 and CD4 + T lymphocytes from uninfected SIVmac239 cells on the growth of cynomolgus monkey B lymphocytes collected at different time points. 3Hthymidine uptake assay The supernatant could inhibit the DNA synthesis of B lymphocytes. The mechanism of apoptosis was detected by gel electrophoresis. The expression of Cyclin D1 was detected by Western blot. Results: The supernatant of CD4 + T lymphocytes infected with SIVmac239 contained cytokines that inhibited the growth of cynomolgus monkey B lymphocytes, and its inhibitory effect increased with time. These factors can inhibit the DNA synthesis of CD4 + T lymphocytes, the highest inhibition rate of up to 81%. Gel test results showed no DNA ladder. Western blot showed that Cyclin D1 expression was inhibited. Conclusion: Infection of CD4 + T lymphocytes by SIVmac239 produces a factor that inhibits the growth of B lymphocytes. This factor can be strongly inhibited the synthesis of DNA of monkey B lymphocytes, but did not cause apoptosis of B lymphocytes. Its main mechanism is to inhibit the production of cyclin D1 in cynomolgus B lymphocytes and stop B lymphocytes in G1 phase. The main mechanism of inhibition is that cells are captured in the G1 phase of the cell cycle. The results of this study found that a new phenomenon is that while the monkey B lymphocytes are not infected by SIVmac239, they can be captured by cytokines produced by CD4 + T lymphocytes infected with SIVmac239 and inhibited their cellular DNA synthesis, Function barriers and affect the body’s role in humoral immunity.