γδT细胞、NK细胞及LAK的部分抗肿瘤生物特性比较

Analysis comparatively the characteristics of γδT cell and LAK

目的 :通过体外分离、增殖培养获取γδT细胞、NK细胞和LAK细胞 ,并比较了 3种细胞的抗肿瘤的生物学特性。方法 :收集用不同单抗分别包装被粘附的细胞 ,然后通过MACS细胞分选仪的分选 ,获取的细胞进行细胞增殖动力学、细胞表型、细胞杀伤活性的测定以单抗阻断效应的分析。结果 :经MACS分离得到的γδT细胞 ,培养 2w后细胞数扩散 6 0 0～ 80 0倍 ,CD3、CD8和γδ细胞表达阳性率分别是 72 2 9%、5 8 0 2 %和 6 5 98%。γδT细胞对NK敏感细胞K5 6 2以及NK不敏感细胞Raji和XG 7这 3种不同靶细胞均有较高的杀伤率 ,分别为 35 98%、42 2 7%和 6 9 0 8% ;NK细胞对此 3种细胞的杀伤率分别是45 12 %、12 34%和 2 9 2 7% ;LAK细胞的杀伤率分别为 44 0 1%、2 9 2 7%和 2 5 6 8%。γδT细胞对经MHC Ⅰ类单抗阻断前后的K5 6 2、Raji和XG 73种靶细胞的杀伤率无明显改变。结论 :γδT细胞、NK细胞和LAK细胞都具有一定的非特异性杀伤肿瘤细胞的作用 ;γδT细胞对MHC Ⅰ类单抗阻断后的肿瘤细胞的杀伤无明显变化 。

Objective:As the function of γδT cells and NK in the immunological therapy is showing more and more important,the characteristics of γδ T cells,NK and LAK were analyzed comparatively after that they were richened by isolating and incubating in vitro.Methods:The cells were collected using MACS after the cells were panned respectively with special monoclonal antibodies.Then the characteristics including proliferation,phenotype,cytotoxin and the down regulation blocked by specific antibodies were analyzed.Results:The γδ T cells can expand 600～800 times after culturing 2 weeks and the percent of CD3,CD8 and γδ expressed on the collected cells were 72.29%,58.02% and 65.98% respectively γδ T cells showed the high cytotoxin to K562(NK sensitive cell line),Raji and XG 7 cell lines(NK non sensitive cell lines).The percentage of cytotoxin reached 35.98%,52.27% and 69.08% respectively compared with γδT respectively.No obviously change of percent of γδT cytotxic ability to these target cells were observed using special MHC class I monoclonal antibody to block the γδT cell before coculturing the target cells with γδ T cells.Conclusion:All of γδ T cells,NK and LAK showed the non specific cytotoxin to tumor cells.The cytotoxic capability of γδ T cells did not be effected after blocking with MHC class I monoclonal antibody.These results demonstrated that γδ T cell could kill more kinds of tumor cells than NK and LAK.