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目的:观察单核细胞、未成熟和成熟树突状细胞(DCs)表达MHCI类相关抗原A(MICA)的情况,以及DCs表达MICA后对NK细胞活性的影响。方法:用流式细胞术分别检测单核细胞、未成熟DC(iDCs)以及LPS、TNF-α、CD40L、IL-15和IFN-α分别刺激的成熟DCs表面MICA的表达,并观察DCs表达MICA后对NK细胞表达CD69、细胞毒活性和分泌IFN-γ的影响。最后用流式细胞术检测重组NKG2D/Fc蛋白和抗IL-12单克隆抗体(mAb)对DCs激活NK细胞的影响。结果:单核细胞不表达MICA,iDCs低表达MI-CA。LPS、TNF-α和CD40L对DCs表达MICA无影响;但IFN-α和IL-15可促进DC上调MICA表达。DCs表达MICA后可促进NK细胞表达CD69、分泌IFN-γ、杀伤K562细胞。NKG2D/Fc蛋白可抑制NK细胞的杀伤活性和分泌IFN-γ;而抗IL-12mAb仅抑制IFN-γ分泌。结论:MICA在DCs表面的表达受局部微环境影响,且DCs表达MICA后可增强NK细胞的活性。
OBJECTIVE: To observe the expression of MICA-associated antigen A (MICA) in monocytes, immature and mature dendritic cells (DCs) and the effect of DCs expressing MICA on NK cell activity. Methods: The MICA expression in monocytes, immature DCs (iDCs) and the matured DCs stimulated by LPS, TNF-α, CD40L, IL-15 and IFN-α were detected by flow cytometry and the expression of MICA On the expression of CD69 on NK cells, the cytotoxic activity and the secretion of IFN-γ. Finally, the effects of recombinant NKG2D / Fc protein and anti-IL-12 monoclonal antibody (mAb) on DCs activated NK cells were detected by flow cytometry. Results: Monocytes did not express MICA, and iDCs had low expression of MI-CA. LPS, TNF-α and CD40L had no effect on the expression of MICA in DCs; however, IFN-α and IL-15 promoted the upregulation of MICA expression in DCs. DCs expression of MICA can promote NK cells to express CD69, secreting IFN-γ, killing K562 cells. NKG2D / Fc protein inhibits NK cell cytotoxic activity and secretes IFN-γ; whereas anti-IL-12 mAb only inhibits IFN-γ secretion. CONCLUSION: The expression of MICA on the surface of DCs is affected by the local microenvironment, and the expression of MICA in DCs can enhance the activity of NK cells.