IL-2促进PBMC来源的NK、NKT、CD8^+ T细胞高表达NKG2D  

Expression pattern of NKG2D on different subsets of high doses IL-2 activated human PBMC

作　　者：邬鹏[1] 魏海明[1] 田志刚[1] 

机构地区：[1]中国科学技术大学生命科学学院免疫学研究所,合肥230027

出　　处：《中华微生物学和免疫学杂志》2006年第8期684-687,共4页Chinese Journal of Microbiology and Immunology

基　　金：国家杰出青年科学基金资助(30125038);国家自然科学基金资助(30371308)

摘　　要：目的观察大剂量IL-2活化的人外周血单个核细胞(PBMC)中,NKG2D在NK细胞、T细胞和NKT细胞表面的表达规律。方法使用三重免疫荧光标记的流式细胞术检测NKG2D的表达情况。使用sMICA蛋白与人PBMC共同培养,之后使用流式细胞术分析NKG2D在NK细胞中的表达情况。使用半定量RT-PCR方法检测大剂量IL-2活化的人PBMC中NKG2D及其锚定蛋白DAP10 mRNA的表达变化。结果使用大剂量IL-2活化人PBMC细胞后,NKG2D在NK细胞、CD8+T细胞和NKT细胞表面的表达均增加,但是在CD4+T细胞表面始终不表达。同时IL-2可以拮抗sMICA对NKG2D的下调作用。半定量RT-PCR结果显示,使用大剂量IL-2活化人PBMC之后,NKG2D及其锚定蛋白DAP10的mRNA水平并不发生明显变化。结论大剂量IL-2培养人PBMC之后,NKG2D在NK细胞、CD8+T细胞和NKT细胞表面的表达均增加,可能是PBMC活化并获得广谱抗肿瘤效应的机制之一。Objective To investigate the expression pattem of NKG2D on different subsets of high doses IL-2 activated human PBMC. Methods Expression of NKG2D on high doses IL-2 activated human PBMC was detected using triple fluorescent staining and flow cytometry analysis. Expression of NKG2D was also detected after PBMC were treated with sMICA and with or without IL-2 respectively. To further investigate the function of NKG2D in killing of tumor cells by PBMC treated with IL-2, levels of mRNA of NKG2D and its adaptor protein DAP10 were detected using semi-quantitive RT-PCR. Results The expression of NKG2D was up-regulated on NK cells, CD8^＋ T cells and NKT cells, but not on CD4^＋ T cells of human PBMC treated with high doses IL-2. After being treated with sMICA protein, the expression of NKG2D on human PBMC was down-regulated. Alternatively, when treated with IL-2, the effect of sMICA on NKG2D expression could be eliminated. Although the expression of NKG2D on different subsets of PBMC could be up-regulated after being treated with IL-2, similar levels of mRNA of NKG2D and its adaptor protein DAP10 was detected in PBMC treated with or without IL-2. Conclusion The expression of NKG2D was up-regulated on NK cells, CD8^＋ T cells and NKT cells of high doses IL-2 activated human PBMC, which could explain the high cytotoxity of such cells in the process of cancer treatment. Meanwhile, the up-regulation effect of IL-2 on NKG2D expression could counteract the effect of sMICA which could down-regulate the NKG2D expression in cancer patients and in experiment in vitro to escape the immunosurveillanee.

关 键 词：外周血单个核细胞 NKG2D NK细胞 CD8^+T细胞 DAP10 

分 类 号：R392[医药卫生—免疫学]