直接和间接共培养条件下小鼠子宫自然杀伤细胞与树突状细胞相互作用的比较

Comparison of interactions between direct and indirect cultured uterine natural killer cells and dendritic cells of mice

背景:子宫自然杀伤细胞与树突状细胞可以相互作用,但相互间作用是否必须直接接触还存在不同的观点。目的:对比直接和间接共培养条件下子宫自然杀伤细胞与树突状细胞之间的相互作用,明确两者相互作用的方式。方法:将树突状细胞和子宫自然杀伤细胞进行直接接触共培养,设为直接共培养组;在2种细胞Transwell系统中进行共培养,设为间接共培养组。观察培养细胞的生长状况,酶联吸附法检测培养上清白细胞介素10,12、转化生长因子β细胞因子的浓度,流式细胞术检测各组细胞表面标志CD86的表达情况。结果与结论:与单独培养树突状细胞和子宫自然杀伤细胞相比,直接和间接共培养组培养液上清中白细胞介素10,12、转化生长因子β浓度均增加(P<0.05),尤以白细胞介素10浓度增加明显(P<0.05),CD86的表达明显增高(P<0.05)。直接共培养组白细胞介素10,12和转化生长因子β浓度更高(P<0.05),表达CD86的细胞含量更多(P<0.05)。证实子宫自然杀伤细胞可以促进树突状细胞成熟,而树突状细胞促进子宫自然杀伤细胞的分泌,两者通过细胞间的直接接触而相互作用。

BACKGROUND：It has been known that uterine natural killer cells and dendritic cells have crosstalk, but the way of theinteractions is controversial.OBJECTIVE：To contrast the interactions between direct and indirect cultured uterine natural killer cells and dendriticcells of mice and then to investigate the way of their interaction.METHODS： Dendritic cells and uterine natural killer cells were co-cultured via direct contact （direct co-culture group）.Dendritic cells and uterine natural killer cells were cultured in Transwell system （indirect co-culture group）. Cellular morphology was observed. The concentrations of interleukin-10, interleukin-12 and transforming growth factor-β in supernatant were detected by enzyme-linked immunosorbent assay, The expressions of cell surface markers CD86 was detected by flow cytometry.RSULTS AND CONCLUSION：Compared with dendritic cells and uterine natural killer cells, the concentrations of interleukin-10, interleukin-12 and transforming growth factor-β in the supernatant of co-culture groups were greatly increased （P 〈 0.05）, especially the levels of the interleukin-10 （P 〈 0.05）, the expression of CD86 of co-culture groups were also increased （P 〈 0.05）. The concentrations of interleukin-10, interleukin-12 and transforming growth factor-β in the supernatant and the expression of CD86 of direct co-culture group were higher than those of indirect co-culture group （P 〈 0.05）. Uterine natural killer cells can promote the maturity of dendritic cells, and dendritic cells are able to facilitate the secretion of uterine natural killer cells. The interaction way is depended on cell-cell contact.