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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 ,抑制其对肿瘤细胞的细胞毒效应
Objective: To analyze the effect of anti-NKG2D polyclonal antibody (pAb) on NK and LAK cytotoxicity. Methods: Peripheral blood mononuclear cells (PBMCs) were isolated by density gradient centrifugation. LAK cells were induced by 10 mg / L LPHA and 1 × 10 6 U / L rhIL 2. The NK cells were sorted by flow cytometry (FCM) Type testing. After adding NKG2DpAb to block the NKG2D molecules on the surface of NK and LAK cells, MTT assay was used to detect the cytotoxic effect. Results: Confirmed by FCM analysis, high purity and highly active NK cells were obtained. Anti-NKG2DpAb can significantly inhibit the cytotoxic effect of NK and LAK cells on K562 and HepG2 cells. The cytotoxic effects of NK cells on both target cells decreased by 82.9% and 75.6%, respectively. The cytotoxic effects of LAK cells on both target cells decreased by 52.8% and 52.2%, respectively. However, anti-NKG2DpAb did not significantly inhibit the cytotoxic effect of both effector cells on human nasopharyngeal carcinoma cell line CNE. Conclusion: Anti-NKG2DpAb can inhibit the cytotoxicity of NKG2DpAb on tumor cells by blocking the NKG2D molecules on the surface of NK and LAK cells