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目的探讨Toll样受体4(TLR4)在β2糖蛋白I(β2GPI)诱导小鼠骨髓来源树突状细胞(BMDC)成熟过程中的作用。方法体外联合使用重组小鼠粒细胞巨噬细胞集落刺激因子(rmGM-CSF)和重组小鼠白细胞介素-4(rmIL-4)诱导TLR4正常(C3H/HeN)及TLR4缺陷(C3H/HeJ)小鼠的骨髓细胞为未成熟树突状细胞(iDC)。用β2GPI或LPS(阳性对照)对iDC进一步刺激后,采用倒置显微镜观察细胞形态,流式细胞术分析细胞表面分子变化,ELISA测定细胞因子IL-12和TNF-α的分泌水平。结果同C3H/HeJ小鼠相比,C3H/HeN小鼠iDC经β2GPI或LPS刺激后,细胞突起较多,形态上更成熟;细胞表面分子CD11c和MHC-Ⅱ的表达量更高(P<0.05);细胞因子IL-12和TNF-α的分泌更强(P<0.01)。结论 TLR4在β2GPI诱导小鼠骨髓细胞来源的树突状细胞成熟过程中起重要作用。
Objective To investigate the role of Toll-like receptor 4 (TLR4) in the maturation of mouse bone marrow-derived dendritic cells (BMDC) induced by β2-glycoprotein I (β2GPI). Methods Recombinant murine granulocyte-macrophage colony stimulating factor (rmGM-CSF) and recombinant murine interleukin-4 (rmIL-4) were used in combination to induce TLR4 deficiency (C3H / HeN) Mice bone marrow cells are immature dendritic cells (iDCs). After stimulation of iDC with β2GPI or LPS (positive control), the cell morphology was observed by inverted microscope. The changes of cell surface molecules were analyzed by flow cytometry. The levels of cytokines IL-12 and TNF-α were measured by ELISA. Results Compared with C3H / HeJ mice, the iDCs of C3H / HeN mice showed more protuberances and more mature morphology after stimulated by β2GPI or LPS, and the expressions of CD11c and MHC-Ⅱ were higher (P <0.05 ); Secretion of cytokines IL-12 and TNF-α was stronger (P <0.01). Conclusion TLR4 plays an important role in the maturation of dendritic cells induced by β2GPI in mouse bone marrow cells.