不同刺激剂和培养环境对CD4+ 、 CD8+ T细胞内细胞因子表达的影响
流式细胞术;,T细胞亚群;,细胞因子;,刺激剂;,培养条件,,]流式细胞术;,T细胞亚群;,细胞因子;,刺激剂;,培养条件,不同刺激剂和培养环境对CD4+、CD8+T细胞内细胞因子表达的影响,1材料和方法,2结果,3讨论,参考文献:
Cytokine expression of CD4+ and CD8+ T cells induced by different stimuli under different culture conditionsZHOU Maohua, WU Changyou*
Department of Immunology, Medical School, Sun Yatsen University; Key Laboratory for Tropical Diseases Control Ministry of Education Guangzhou 510080, China
[Abstract] AIM: To investigate the effect of different stimuli and different culture conditions on the cytokine expression of CD4+ and CD8+ T cells. METHODS: PBMCs were isolated from normal human peripheral blood and cultured with three kinds of stimuli (PHA, antiCD3 and antiCD28 mAbs, PMA and ionomycin) under four different culture conditions (room temperature, 37℃ water bath, 37℃ incubator, 37℃ and 50 mL/L CO2 incubator) for 4.5~5 h and intracellular cytokines (IL2, IFNγ and TNFα) in T cells were assessed by flow cytometry (FCM). RESULTS: Cytokine expression of CD4+ and CD8+ T cells varied when treated with different stimuli under different conditions. PMA had the strongest stimulative effect on cytokine expression of PBMC, while the effect of antiCD3 mAb was weaker, and that of PHA was the weakest. Different culture conditions did not greatly change the cytokine expression profile of T cells (P>0.05) except that there were very few cytokine producing cells when PBMCs were cultured at room temperature. CONCLUSION: AntiCD3 and antiCD28 mAbs, PMA and ionomycin are recommended for the stimulation of PBMCs and detection of intracellular cytokine using FCM. Culture temperature but not the concentration of CO2 is the most important factor during the short term T cell stimulation. ......
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