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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。
OBJECTIVE: To clarify the subpopulation and anatomical localization of natural keratin-responsive B cells and to analyze their ability to secrete natural anti-keratinutoantibody (AKautoAb). Methods: The splenocytes and peritoneal cells of SPF grade C57BL / 6 mice were stained and stained with fluorescent antibody. The keratin-responsive B lymphocyte subsets were analyzed by flow cytometry. After spleen and peritoneal lymphocytes were cultured in vitro, the titer of AKautoAb was analyzed by ELISA, and the number of B cells secreting AKautoAb was analyzed by ELISPOT. RESULTS: Almost all keratin-binding B cells in the peritoneal cavity were B-1a cells, and keratin-binding B cells in the spleen were predominantly marginal B cells. The number of cells secreting AKautoAb in peritoneal cells was significantly more than that in spleen cells, and the titer of AKautoAb in the culture supernatant was significantly higher than that of splenocytes. CONCLUSION: Keratin-responsive B cells exist in three mature B cell subsets: B-1 cells, follicular B cells and marginal B cells, of which CD5 + B-1a cells have an active ability to secrete AKautoAb.