抗NKG2D多克隆抗体抑制NK和LAK细胞细胞毒效应的研究

Study of the inhibiting effect of anti-NKG2D polyclonal antibody on cytotoxicities of NK and LAK cells

目的 :分析抗NKG2D多克隆抗体 ( pAb)对NK和LAK细胞毒作用的影响。方法 :应用密度梯度离心法分离外周血单个核细胞 (PBMC) ,经 10mg/LPHA和 1× 10 6U/LrhIL 2诱导LAK细胞产生 ,再应用流式细胞术 (FCM)分选NK细胞并进行表型检测。加入抗NKG2DpAb封闭NK和LAK细胞表面的NKG2D分子后 ,用MTT比色法检测其细胞毒效应。结果 :经FCM分析证实 ,获得高纯度、高活性的NK细胞。抗NKG2DpAb能显著抑制NK和LAK细胞对K5 6 2、HepG2细胞的细胞毒效应。NK细胞对两种靶细胞的细胞毒效应分别下降了 82 .9%和 75 .6 % ;LAK细胞对两种靶细胞的细胞毒效应分别下降了 5 2 .8%和 5 0 .2 %。但抗NKG2DpAb不能显著抑制两种效应细胞对人鼻咽癌细胞系CNE的细胞毒效应。结论 :抗NKG2DpAb可通过封闭NK和LAK细胞表面的NKG2D分子 。

AIM: To investigate the effect of anti NKG2D polyclonal antibody(pAb) on cytotoxicities of NK and LAK cells. METHODS: Peripheral blood mononuclear cells(PBMCs) were separated by centrifugation on Ficoll Hypaque gradients. LAK cells were induced from PBMCs by PHA (10 mg/L) and rhIL 2 (1×10 6 U/L ). Then NK cells were sorted by flow cytometry(FCM). The cytotoxicities of NK and LAK cells were analyzed by MTT colorimetry after NKG2D molecule on NK and LAK cells were blocked with anti NKG2D pAb. RESULTS: FCM analysis proved that both purity and activity of obtained NK cells were high.The anti NKG2D pAb could inhibit significantly cytotoxicities of NK and LAK cells to K562 and HepG2 cells, for NK cells,having decreased 82.9% and 75.6%, for LAK cells,having decreased 52.8% and 50.2%, respectively.The anti NKG2D pAb had no effect on cytotoxicities of NK and LAK cells to CNE cells. CONCLUSION: The anti NKG2D pAb can inhibit cytotoxocities on tumor cells by NK and LAK cells through NKG2D molecule on two effector cells.