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目的 :建立一种特异快速地诱导扩增人外周血γδT细胞的方法 ,并比较了γδT细胞 ,NK和LAK细胞的抗肿瘤的生物学特性。方法 :收集用不同单抗分别包被粘附的细胞 ,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及单抗阻断效应的分析。结果 :经MACS分离得到的γδT细胞 ,培养 2周后细胞数扩增 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% ,5 2 .2 7%和 6 9.0 8% ;NK细胞对此 3种细胞的杀伤率分别是 45 2 1% ,12 .34%和 11.94% ;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。提示γδT细胞较NK细胞和LAK细胞有更广泛的抗瘤谱
OBJECTIVE: To establish a method to specifically and rapidly induce human peripheral blood γδT cells, and to compare the anti-tumor biological characteristics of γδT cells, NK cells and LAK cells. Methods: The cells that were coated with different McAbs were collected, and then the cell proliferation kinetics, cell phenotype and cytocidal activity were determined by the sorting of MACS cell sorter, and the monoclonal antibody blocking effect Analysis. Results: The number of γδT cells isolated by MACS was 600 ~ 800 times higher after 2 weeks of culture, and the positive rates of CD3, CD8 and γδ were 72.29%, 58.02% and 65% respectively .98%. γδT cells had higher killing rates on NK sensitive cell K5 6 2 and NK-sensitive cells Raji and XG 7, which were 35.98%, 52.27% and 6 9.0 8% respectively, ; NK cell killing rates of these three kinds of cells were 45 2 1%, 12.34% and 11.94% respectively; killing rates of LAK cells were 44.0%, 2 9.27% and 25.56% . The killing rate of γδT cells to K562, Raji and XG 73 kinds of target cells before and after the blocking by MHC class I McAb was not significantly changed. CONCLUSION: γδT cells, NK cells and LAK cells all have a certain non-specific killing effect on tumor cells; γδT cells have no significant change on the killing of tumor cells after MHC class I antibody blocking. This suggests that γδT cells have a broader anti-tumor spectrum than NK cells and LAK cells