摘要
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γδT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γδT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 au- tologous 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 γδ T cells was as- sessed by Lactate dehydrogenase （LDH） assay. Results The percentages of HIV-P24 ＋ CIM ＋ T cells reached the peak at 10 days.γδ T cells from AIDS patients can kill not only the autologous HIV-infected CD4 ＋T cell, hut also the HIV uninfected CD4 ＋T cells （the cytolysis rate were 71.1 ±9.7 and 24.2 ± 18.2,respectively）. However, γδ T cells from healthy donors could not kill the autologous CD4 ＋ T cells （the eytolysis rate was 4.3 e 10. 1 ）.Conclusions γδ 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 γδ T cells adoptive immunotherapy.
出处
《基础医学与临床》
CSCD
北大核心
2012年第7期778-782,共5页
Basic and Clinical Medicine
基金
国家"十二五"重大专项(2012ZX10001-006)
国家重点基础研究发展规划(973计划)(2006CB504205)
