目的：观察中国猕猴的CD4+CD25+调节性T细胞(Tregs)对Vγ2Vδ2 T细胞体外增殖的影响。方法：取3只正常的无潜伏感染的中国猕猴和6只3个月内人工感染过卡介苗的中国猕猴，分离9只动物外周血单个核细胞(PBMCs)，免疫磁珠法分选纯化出Tregs。将去除Tregs的PBMCs标记CFSE后，再与Tregs混合，给予CD3/CD28抗体、HMBPP和(或)IL-2刺激，培养7 d，流式细胞术检测Vγ2Vδ2 T细胞的增殖情况。从3只正常中国猕猴分离PBMCs，经过IL-2联合HMBPP体外刺激5 d后，标记上CFSE，再与当天新鲜分离的相同动物的Tregs混合，在IL-2刺激下继续培养4 d后，流式细胞术检测Vγ2Vδ2 T细胞的增殖情况。结果：Tregs不仅抑制Vγ2Vδ2 T细胞的HMBPP抗原特异性扩增以及CD3/CD28抗体诱导的抗原非特异性扩增(t=2.571，10.430，P＜0.05)，而且以剂量依赖关系抑制IL-2联合HMBPP体外激活的Vγ2Vδ2 T细胞(F=9.880，P＜0.001)。IL-2联合HMBPP体外预激活的Vγ2Vδ2 T细胞的扩增仍然能够受到Treg的显著抑制(FTregs=29.800，FIL-2=59.290，F交互=30.410，P均＜0.001)。结论：中国猕猴的Tregs对Vγ2Vδ2T有直接的体外负调节作用。%Aim: To investigate the effect of rhesus macaques CD4 * CD25 * regulatory T cells (Tregs) on Vγ2VS2 T cells in vitro. Methods: CFSE-labeled, CD4 * CD25 * T cells-depleted PBMCs, which were separated from the peripheral blood of 6 BCG-infected rhesus macaques, were cultured alone or co-cultured with indicated ratios of purified Tregs for 7 d in the stimulation of purified CD3, CD28 antibodies, HMBPP with/without IL-2 in dividually. CFSE-labeled PBMCs, which had been separated from the peripheral blood of 3 naive rhesus macaques and stimulated with HMBPP plus IL-2 for 5 days, were co-cultured with different amounts of fresh Tregs purified from the peripheral blood of the same animals for another 4 days in the stimulation of IL-2. The proliferations of the CFSE-labeled Vγ2V52 T in PBMCs was analyzed by flow cytome-try. Results: Tregs could suppress the HMBPP-specific and CD3/CD28 antibody-induced non-specific proliferations of Vγ2Vδ2 T cells (t = 2.570,10.430, P <0.05), Tregs have the function of suppressing the IL-2 plus phosphoantigen induced activations/proliferations of Vγ2Vδ2 T cells in a dose-dependent manner in vitro ( F = 9. 880, P < 0. 001). Tregs suppressed the sequent proliferations of Vγ2V52 T cells in PBMCs pre-activated by IL-2 plus HMBPP( Ftrega =29. 800, FIL-2 = 59.290,FmKaeSo = 30.410,P<0.001). Conclusion:Tregs of the rhesus macaques have direct negative regulatory effect on V-y2V82 T cells in vitro.
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