天然角蛋白自身反应性B细胞亚群及功能的分析

Studies on subset, location and secretion of natural keratin-reactive B cells

目的:明确天然角蛋白反应性B细胞的亚群及解剖定位,初步分析其分泌天然抗角蛋白自身抗体(anti-keratinautoantibody,AKautoAb)的能力。方法:取SPF级C57BL/6小鼠的脾细胞和腹腔细胞,荧光抗体染色后用流式细胞仪分析角蛋白反应性B细胞亚群。将脾脏和腹腔淋巴细胞体外培养后,用ELISA分析AKautoAb的滴度,用ELISPOT法分析分泌AKautoAb的B细胞数。结果:腹腔中几乎所有结合角蛋白的B细胞均为B-1a细胞,脾脏中结合角蛋白的B细胞以边缘带B细胞为主。腹腔细胞中分泌AKautoAb的细胞数显著多于脾细胞,其培养上清中AKautoAb的滴度显著高于脾细胞。结论:角蛋白反应性B细胞存在于3个成熟B细胞亚群:B-1细胞、滤泡B细胞和边缘带B细胞,其中CD5+的B-1a细胞具有活跃的分泌AKautoAb的能力。

AIM： To clarify the subsets and location of B cells responsible for the production of natural anti-keratin autoantibody （AK auto Ab）, and analyze the function of each subset. METHODS： Cells isolated from spleen and peritoneal cavity of C57BL/6 mice were cultured in vitro. FACS was employed to detect keratin reactive B cell subsets. The anti-keratin IgM level in supernatant was analyzed by ELISA, while ELISPOT was utilized to detect the B cells which can secrete AK auto Ab. RESULTS： Keratin reactive B cells were mainly marginal zone B cells in spleen, while they were mainly B-1 a cells in peritoneal cavity. There were more B cells producing AK auto Ab in peritoneal cavity than in spleen. The anti-keratin IgM level in the supernatant of cultured peritoneal cells was much higher than that of splenocytes, CONCLUSION： There are three subsets of keratin-reactive B cells： B-1 B cells, follicular B cells and marginal zone B cells. B-1 B cells are active in secreting AK auto Ab.