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目的:体外观察中国猕猴CD4+CD25+调节性T细胞(Tregs)对T淋巴细胞产生的抗PPD抗原特异性扩增的影响。方法:分离6只3月内人工感染过结核分枝杆菌卡介苗的雄性中国猕猴外周血单个核细胞(PB-MCs),免疫磁珠法分选纯化出Tregs,并用PKH26红标记。将去除Tregs的剩余PBMCs标记上CFSE,然后单独或与纯化的Tregs混合,分别给予结核杆菌纯化蛋白衍生物(PPD)或CD3/CD28单克隆抗体刺激,体外培养7d,用流式细胞术分析含或不含Tregs的PBMCs中CD4+T淋巴细胞、CD8+T淋巴细胞和Vγ2Vδ2T淋巴细胞对PPD抗原和CD3/CD28抗体刺激的反应。结果:PPD不仅能引起CD4+T淋巴细胞、CD8+T淋巴细胞扩增,而且引起Vγ2Vδ2T淋巴细胞也发生扩增。Tregs不仅能抑制CD3/CD28抗体诱导的CD4+T淋巴细胞、CD8+T淋巴细胞和Vγ2Vδ2T淋巴细胞的抗原非特异性扩增,而且还可抑制PPD抗原刺激引起的CD4+T淋巴细胞、CD8+T淋巴细胞和Vγ2Vδ2T淋巴细胞的抗原特异性扩增。结论:中国猕猴Tregs对T淋巴细胞产生的抗结核杆菌特异性免疫反应有负调节作用。
Objective: To observe the effects of CD4 + CD25 + regulatory T cells (Tregs) on the specific amplification of anti-PPD antigen produced by T lymphocytes in Chinese macaques. METHODS: Six PBMCs were isolated from 3 Chinese macaques BCG-infected macaques in March. The PBMCs were isolated by magnetic activated cell sorting and labeled with PKH26. The remaining PBMCs from Tregs were labeled with CFSE and then mixed with purified Tregs alone or in combination with purified Mycobacterium tuberculosis purified protein derivative (PPD) or CD3 / CD28 monoclonal antibody. The PBMCs were cultured in vitro for 7 days and analyzed by flow cytometry Or CD4 + T lymphocytes, CD8 + T lymphocytes and Vγ2 Vδ2 T lymphocytes in Tregs-depleted PBMCs against PPD antigen and CD3 / CD28 antibody stimulation. Results: PPD not only caused the expansion of CD4 + T lymphocytes and CD8 + T lymphocytes, but also caused the expansion of Vγ2Vδ2T lymphocytes. Tregs can not only inhibit the non-specific amplification of antigen of CD4 + T lymphocytes, CD8 + T lymphocytes and Vγ2Vδ2T lymphocytes induced by CD3 / CD28 antibody, but also inhibit the CD4 + T lymphocytes stimulated by PPD antigen, CD8 + T Antigen-specific amplification of lymphocytes and Vγ2Vδ2T lymphocytes. CONCLUSION: Tregs from Chinese macaques have a negative regulatory effect on T-lymphocyte-specific anti-tuberculosis immunoreactivity.