波形蛋白的警报素功能鉴定

目的：鉴定胞外可溶性波形蛋白（Vim）促进相关细胞增殖、活化、趋化的能力。方法：体外检测Vim刺激大鼠脾细胞的增殖情况。体内检测Vim免疫小鼠的外周血淋巴细胞数及Vim抗体水平。收集小鼠腹腔巨噬细胞与不同浓度Vim共培养，检测其对巨噬细胞吞噬鸡红细胞能力的影响。以Transwell小室法检测Vim对NIH3T3成纤维细胞的趋化作用。结果：体外实验Vim剂量依赖性地促进大鼠脾细胞增殖，16 μg/ml浓度的Vim刺激预致敏或未致敏脾细胞增殖率分别高达（196.0±9.7）%、（208.9±4.6）%，且该二者无显著性差异。体内实验单用Vim免疫小鼠的外周血淋巴细胞浓度（10.9 L -1 ）激增（5.74±0.51 vs 1.69±0.13），同时激发了Vim特异性抗体水平（OD值2.31±0.06 vs 0.19±0.08）；且与Vim辅以佐剂免疫小鼠的相应指标均无显著性差异。体外实验Vim浓度依赖性地促进巨噬细胞吞噬鸡红细胞的能力，并能够对NIH3T3成纤维细胞产生趋化作用。结论：胞外可溶性Vim非特异性促进炎症反应相关细胞增殖、活化、趋化，具有警报素功能。

Functional verification of vimentin as an alarmin

  Objective: To identify the function of extracellular soluble vimentin that promotes proliferation,activation and chemotaxis of inflammatory cells. Methods: The proliferation of rat splenocytes stimulated with vimentin was evaluated in vitro.The lymphocyte counts and vimentin antibody levels of peripheral blood in mice immunized with vimentin were detected in vivo.Peritoneal macrophages were collected and cultured with different concentrations of vimentin to detect the effect on phagocytosing chicken red blood cells.The chemotaxis of NIH3T3 fibroblast towards vimentin was observed in transwell chamber. Results:  In vitro vimentin dose dependently promoted the proliferation of splenocytes.The proliferation indexes of primed and naive splenocytes cultured with 16 μg/ml vimentin were reached up to(196.0±9.7)% and(208.9±4.6)% respectively without significant difference.In vivo vimentin significantly enhanced the lymphocytes number(10.9 L -1 )of peripheral blood(5.74±0.51 vs 1.69±0.13)and the levels of vimentin specific antibody(OD value 2.31±0.06 vs 0.19±0.08)that shown no significant difference from immunization with vimentin plus CFA. In vitro vimentin dose dependently stimulated phagocytic ability of macrophages and performed the chemotactic effects on NIH3T3 fibroblasts. Conclusion:  Extracellular soluble vimentin promotes the proliferation,activation and chemotaxis of concerned inflammatory cells and possesses the functions as an alarmin.