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目的研究3种分枝杆菌耐热蛋白抗原体外对人γδT细胞的刺激活性。方法分离健康人外周血单个核细胞(PBMC),分别使用人型结核分枝杆菌H37Ra耐热蛋白抗原(H37Ra-HAg)、牛型结核分枝杆菌BCG耐热蛋白抗原(BCG-HAg)和耻垢分枝杆菌耐热蛋白抗原(MS-HAg)进行刺激并持续培养12d,采用流式细胞仪检测增殖细胞中γδT细胞数量及CD4+T细胞所占比例。结果 H37Ra-HAg刺激组扩增细胞中γδT细胞的比例和数量均显著高于BCGHAg、MS-HAg刺激组和IL-2对照组[((50.52±5.40)%v.s(7.31±3.15)%、(5.82±3.42)%、(5.56±2.68)%);(9.63±1.64)×106v.s(0.69±0.36)×106、(0.57±0.22)×106、(0.34±0.18)×106))]。BCG-HAg刺激组扩增细胞中CD4+T细胞的比例和数量均显著高于H37Ra-HAg刺激组和IL-2对照组[(73.56±7.12%、66.93±7.65)%v.s(23.18±6.29)%、(52.61±5.37)%;(4.75±0.86)×106、(3.28±0.59)×106 v.s(2.71±0.65)×106、(1.63±0.45)×106]。而BCG-HAg刺激组和MS-HAg刺激组扩增细胞中γδT细胞的比例和数量和IL-2对照组相比并无显著性差异。结论 H37Ra-HAg具有促进γδT细胞增殖活性,BCG-HAg和MS-HAg具有促进CD4+T细胞增殖活性,BCG-HAg和MS-HAg无刺激γδT细胞增殖活性。
Objective To study the stimulatory activity of three mycobacterial heat-resistant protein antigens on human γδT cells in vitro. Methods Peripheral blood mononuclear cells (PBMCs) were isolated from human peripheral blood mononuclear cells (PBMCs). Human H37Ra heat-tolerant protein antigen (H37Ra-HAg), Mycobacterium bovis BCG heat-resistant protein antigen Mycobacterium tuberculosis heat-resistant protein antigen (MS-HAg) were stimulated and cultured for 12 days. The number of γδT cells and the proportion of CD4 + T cells in proliferating cells were detected by flow cytometry. Results The proportion and number of γδT cells in the H37Ra-HAg-stimulated group were significantly higher than those in the BCGHAg, MS-HAg and IL-2 control groups [(50.52 ± 5.40) vs (7.31 ± 3.15)%, 5.82 ± 3.42%, (5.56 ± 2.68)%); (9.63 ± 1.64) × 106v.s (0.69 ± 0.36) × 106, (0.57 ± 0.22) × 106, (0.34 ± 0.18) × 106))]. The proportion and number of CD4 + T cells in BCG-HAg-stimulated group were significantly higher than those in H37Ra-HAg-stimulated group and IL-2 control group [(73.56 ± 7.12%, 66.93 ± 7.65)% vs (23.18 ± 6.29) %, (52.61 ± 5.37)%; (4.75 ± 0.86) × 106, (3.28 ± 0.59) × 106 vs (2.71 ± 0.65) × 106, (1.63 ± 0.45) × 106]. However, there was no significant difference in the proportion and number of γδT cells between the BCG-HAg-stimulated group and the MS-HAg-stimulated group compared with the IL-2 control group. Conclusion H37Ra-HAg can promote the proliferation activity of γδT cells. BCG-HAg and MS-HAg can promote the proliferation activity of CD4 + T cells, and the proliferation activity of γδT cells without BCG-HAg and MS-HAg.