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目的:观察固相MHC I类相关抗原A(iMICA)刺激的NK细胞对树突状细胞(DCs)活性的影响。方法:首先取新鲜分离及受固相MICA刺激的异体NK细胞,或IL-2、及IL-2联合iMICA刺激的自体NK细胞与未成熟DCs(iDCs)按5∶1比例孵育24 h后,用流式细胞术(FCM)分析HLA-DR+或CD86+频率的DCs。然后取自体NK细胞以iMICA刺激后,按1∶5的比例与iDCs孵育24 h后,FCM检测DCs上HLA-DR、CD86的表达。最后在NK细胞与DCs的共培养体系中加入抗IFN-γ抗体,观察DCs上HLA-DR、CD86表达的变化。结果:当NK细胞与iDCs孵育比例为5∶1时,异体新鲜分离的与自体活化的NK细胞均能杀伤iDCs;而iMICA无协同作用。当NK细胞与iDCs孵育的比例为1∶5时,iMICA刺激的NK细胞可促进DCs表达HLA-DR和CD86;而加入抗IFN-γ抗体可抑制NK细胞诱导DCs表面HLA-DR和CD86表达的上调。结论:异体新鲜分离的或自体活化的NK细胞杀伤iDCs时,无需iMICA的刺激;但当NK细胞的数目明显低于iDCs时,iMICA可刺激NK细胞分泌IFN-γ促进DCs成熟。
Objective: To observe the effects of NK cells stimulated by solid phase MHC class I antigen (iMICA) on the activity of dendritic cells (DCs). Methods: After freshly isolated and allogeneic NK cells stimulated by solid phase MICA or IL-2 and IL-2 combined with iMICA-stimulated autologous NK cells and immature DCs (iDCs) were incubated in 5: 1 ratio for 24 h, DCs for HLA-DR + or CD86 + frequencies were analyzed by flow cytometry (FCM). Then, NK cells were harvested and stimulated with iMICA. After being incubated with iDCs for 1 h at a ratio of 1: 5, FCM was used to detect the expression of HLA-DR and CD86 on DCs. Finally, anti-IFN-γ antibody was added into the co-culture system of NK cells and DCs to observe the changes of HLA-DR and CD86 on DCs. RESULTS: When NK cells were incubated with iDCs at a ratio of 5:1, allogeneic freshly isolated and autologous NK cells could kill iDCs; however, iMICA had no synergistic effect. When NK cells were incubated with iDCs at a ratio of 1: 5, iMICA-stimulated NK cells could promote the expression of HLA-DR and CD86 in DCs; while the addition of anti-IFN-γ antibody inhibited the expression of HLA-DR and CD86 on the surface of DCs induced by NK cells Increase. CONCLUSION: iMICA stimulation is not required when allogeneic freshly isolated or autologous NK cells kill iDCs. However, when the number of NK cells is significantly lower than that of iDCs, iMICA can stimulate NK cells to secrete IFN-γ to promote the maturation of DCs.