目的 建立自体人类免疫缺陷病毒(HIV)感染的CD4+T细胞模型,检测获得性免疫缺陷综合征(AIDS)患者γδT细胞对自体HIV感染的CD4+T细胞的杀伤作用.方法 分离健康志愿者和未治疗的AIDS患者外周血单个核细胞(PBMCs),磁珠分选获得高纯度的CD4+T细胞,加入植物血凝素(PHA)和白细胞介素-2(IL-2)共同培养,建立自体HIV感染的CD4+T细胞模型.流式细胞术检测HIV-P24+ CD4+T细胞的比例.乳酸脱氢酶(LDH)法测定γδ T细胞的细胞毒作用.结果 培养到第10天时,HIV-P24+ CD4+T细胞的比例达到最高值.AIDS患者的γδT细胞不仅能够杀伤自体HIV感染的CD4+T细胞(细胞溶解率为71.1±97),而且对HIV也有一定的杀伤作用(细胞溶解率为24.2±18.2).而健康志愿者γδT细胞对自体CD4+T细胞没有杀伤作用(细胞溶解率为4.3±10.1).结论 AIDS患者γδ T细胞能够杀伤自体HIV感染的CD4+T细胞,为采用γδT细胞过继免疫治疗AIDS提供了依据.%Objective Establish a model of autologous Human Immunodeficiency Virus ( HIV ) -infected CD4 + T cell, to investigate the cytotoxicity of γδ T cells from acquired immunodeficiency syndrome (AIDS) patients to autologous HIV-infected CD4 + T cells. Methods Peripheral blood mononuclear cells ( PBMCs ) were isolated from healthy donors and AIDS patients. CD4 + T cells with high purity were harvested through negative isolation using magnetic activated cell sorting ( MACS). To establish the autologous HIV-infected CD4+ T cell model, CD4 + T cells from AIDS patients were stimulated with phytohemagglutinin ( PHA) and co-cultured with interleukin-2 (IL-2). Percentages of HIV-P24 + CD4 + T cells were detected by flow cytometry. Cytotoxicity of δγ T cells was assessed by Lactate dehydrogenase (LDH) assay. Results The percentages of HIV-P24 + CD4 + T cells reached the peak at 10 days.δγ T cells from AIDS patients can kill not only the autologous HIV-infected CD4+T cell, but also the HIV uninfected CD4 + T cells (the cytolysis rate were 71. 1 ± 9. 7 and 24. 2 ± 18. 2,respectively). However, 78 T cells from healthy donors could not kill the autologous CD4 + T cells (the cytolysis rate was 4. 3 ± 10. 1).Conclusions δγ T cells of AIDS patients can kill the autologous HIV-infected CD4+T cells, which provide a new sight for the clinical therapy of AIDS based on δγ T cells adoptive immunotherapy.
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