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目的:探讨重组人白细胞介素-17(Interleukin-17,IL-17)对小鼠骨髓造血前体细胞和人脐血来源的CD34~+干细胞生长发育的影响.方法:采用常规方法采集小鼠造血前体细胞;采用Mini-MACS分离技术,从正常人脐血分离人CD34~+干细胞.体外加入IL-17和/或GM-CSF、IL-4培养分离的前体细胞,应用流式细胞仪检测其表型,采用ELISA法检测了其分泌的IL-12水平,通过[~3H]-TdR掺入法测定其刺激同种异体T淋巴细胞增殖的能力.结果:IL-17促进了小鼠骨髓来源的未成熟DC表达Ia,B7-2等免疫分子,促使其分泌较高水平的IL-12,该细胞也能刺激同种异体T细胞有效增殖,表现出了成熟DC的特征.IL-17单独培养9d促使人脐血CD34~+干细胞扩增了2倍,部分细胞高表达CD1a及B7-2,低表达HLA-DR,未检测到CD83的表达.该细胞能促使同种异体T细胞增殖,但作用较弱;而rhIL-17与GM-CSF联合培养后扩增了14倍,培养细胞中CD1a、B7-2阳性细胞的比例明显升高,且此细胞刺激同种异体T细胞增殖的能力较强.结论:IL-17体外可促进小鼠骨髓造血前体细胞来源的DC成熟;与GM-CSF联合培养既能促进CD34~+干细胞增殖,又能使之获得DC特征,初步提示IL-17与GM-CSF联合作用可促进CD34~+干细胞向DC分化.
Objective: To investigate the effect of recombinant interleukin-17 (IL-17) on the growth and development of murine bone marrow-derived precursor cells and human umbilical cord blood-derived CD34 + stem cells.Methods: Mice Hematopoietic precursor cells were isolated from human umbilical cord blood by using Mini-MACS technique, and the isolated precursor cells were cultured with IL-17 and / or GM-CSF in vitro. Flow cytometry The IL-12 level was measured by ELISA, and its ability to stimulate the proliferation of allogeneic T lymphocytes was measured by [~ 3H] -TdR incorporation method.Results: IL-17 promoted small Murine bone marrow-derived immature DCs express Ia, B7-2 and other immune molecules, prompting them to secrete higher levels of IL-12, which can also stimulate effective proliferation of allogeneic T cells and exhibit the characteristics of mature DC. -17 alone induced 9-day-old human umbilical cord blood CD34 ~ + expansion 2-fold, some of the high expression of CD1a and B7-2, low expression of HLA-DR, did not detect the expression of CD83. The cells can promote allogeneic T However, the effect of rhIL-17 on GM-CSF was 14-fold, and the positive rate of CD1a and B7-2 positive cells Cases were significantly increased, and this cell stimulated the proliferation of allogeneic T cells stronger.Conclusion: IL-17 in vitro can promote mouse bone marrow hematopoietic derived DC maturation; combined with GM-CSF can both promote CD34 ~ + Stem cell proliferation, but also to obtain DC characteristics, preliminary suggests that IL-17 and GM-CSF combined action can promote CD34 ~ + stem cells to DC differentiation.