幼年类风湿关节炎活动期血清对树突状细胞细胞周期、凋亡及脾淋巴细胞增殖的影响

目的探讨幼年类风湿关节炎(JRA)活动期血清对树突状细胞(DC)细胞周期、凋亡及脾淋巴细胞增殖的影响,以便进一步探讨DC在JRA发病机制中的作用。方法分离来源于正常小鼠的骨髓细胞,体外用细胞因子(IL-4,GM-CSF)诱导、分化为DC,脂多糖(LPS)刺激获取成熟DC;常规法获取同种异体脾淋巴细胞。光镜观察DC形态改变、流式细胞术对其进行鉴定。利用流式细胞仪分析JRA活动期血清辅助DC成熟过程对DC细胞周期、凋亡产生的影响MTT法检测其对淋巴细胞增殖的影响。结果JRA血清组与正常血清组比较,DC G1期细胞数减少、S G2/M期细胞数显著增多(P<0.05)。DC凋亡细胞数有下降趋势。JRA血清能明显促进淋巴细胞增殖(P<0.05),成熟DC培养液与JRA血清共同作用时上述增殖作用更显著(P<0.01)。结论JRA活动期血清能促进DC和淋巴细胞增殖,同时可能抑制DC凋亡。JRA的发生可能与DC细胞周期、凋亡的调节以及DC介导的免疫放大机制有关。

Effect of Juvenile Rheumatoid Arthritis Active Serum on Cell Cycle, Apoptosis of Dendritic Cell and Cell Proliferation of Splenic Lymphocytes

Objective To investigate the effect of juvenile rheumatoid arthritis(JRA) active phase serum on cell cycle, apoptosis of dendritic cell(DC) and cell proliferation of splenic lymphocyte, so as to approach a mechanism of DC in JRA further. Methods DCs were isolated from healthy murine(Balb/c)bone marrow ,and cultivated with the induction and differentiation of cytokine interleukin-4(IL-4),granulocyte macrophage colony stimulating factor(GM-CSF),and simulated to promote its maturity with lipopolysaccharide(LPS);allogenic splenic lymphocyte was obtained by common method;DC shape was viewed under light microscope.DC were identified about marker and function molecule,and detected the change of cell cycle and apoptosis during DC maturity with JRA serum by flow cytometry. Meanwhile, the influence of lymphocyte proliferation was detected by MTT method.Results JRA serum group,compared with normal serum group,showed that the number of cells in G1 phase decreased, but that in S G2/M period increased (P<0.05).Apoptosis cell population was a downtrend. The lymphocyte proliferation,compared with normal serum group, was obvious(P<0.05),while with the co-effect of DC culture fluid and JRA serum,the proliferation role was more significant than that mentioned above(P<0.01). Conclusions Cell proliferation was augmented and apoptosis may be of DC by JRA serum in active phase.JRA mechanism may be related to immunological regulation of DC cell cycle and apoptosis,as well as immunological amplification mechanism mediated by DC.