骨髓间充质干细胞对异体外周血B淋巴细胞的免疫调节作用  

The immunoregulatory effects of bone marrow mesenchymal stem cells on allogeneic peripheral B lymphocyte

作　　者：武令启[1] 白海[1] 王存邦[1] 杨小亮[1] 赵强[1] 杨义武[1] 林梅[1] 

机构地区：[1]兰州军区总医院血液科暨全军血液学中心,730050

出　　处：《中华微生物学和免疫学杂志》2008年第6期486-491,共6页Chinese Journal of Microbiology and Immunology

基　　金：全军“十一五”杰出人才基金资助项目（06J005）

摘　　要：目的探讨骨髓间充质干细胞（mesenchymal stem cells，MSC）在体外对异体外周血B淋巴细胞的免疫调节作用。方法用密度梯度离心法从骨髓中分离、培养MSC，从外周血中分离单个核细胞，L-亮氨酸甲酯去除单核细胞，以2-氨乙基硫脲溴化物（AET）处理绵羊红细胞（SRBC）的花环形成法，去除T淋巴细胞获得纯化的B淋巴细胞。用羊抗人IgM单克隆抗体（anti-IgM）刺激与或未与MSC或其培养上清共培养3d的B淋巴细胞，应用MTT法测B淋巴细胞的增殖，ELISA法测培养上清中免疫球蛋白IgG、IgM的产生，应用流式细胞术分别检测与MSC共培养24、48h后B淋巴细胞的凋亡。结果MSC及其培养上清抑制由anti-IgM诱导的B淋巴细胞的增殖和分泌IgG、IgM，且随着MSC细胞数量及其培养上清浓度的增加，这种抑制越明显。流式细胞术结果显示，与MSC共培养不同时间的B淋巴细胞的凋亡变化无统计学意义，MSC抑制B淋巴细胞的增殖存在暂时性和可逆性。结论MSC对异体外周血B淋巴细胞存在免疫调节作用，并且这种调控机制是复杂的，不仅与MSC细胞数量有关，还与细胞间的相互作用和MSC分泌的细胞因子有关。Objective To study the immunoregulatory effects of bone marrow mesenchymal stem ceils （MSC） on allogeneic peripheral B lymphocytes in vitro. Methods MSCs were isolated and cultured from bone marrow by gradient centrifugation. Mononuclear ceils were isolated routinely from peripheral blood, then monocytes were eliminated by L-leucine methy ester method. Remained T lymphocytes were eliminated by AET-SRBC rosette method. The action of MSCs and its supernatant on B lymphocytes proliferation in the presence of anti-human IgM goat antibodies （anti-IgM） was investigated by MTT. The IgG, IgM in the supernatant were detected by ELISA. The percent of apoptosis B lymphocytes, co-cultured with MSCs for 24 or 48 h, was assayed by FACS. Results MSCs and its supernatant inhibited B lymphocytes proliferation and Ig secretion. The inhibitory effect depended on the amount of MSCs and condition of its supernatant. The date of FACS indicated that the apoptosis ratio of B lymphocytes, co-cultured with MSCs for different times, were non-significant. The inhibitory effect of MSCs on B lymphocytes was temporary and reversible. Conclusion MSCs have immunoregulatory effects on B lymphocytes, and its mechanisms are complex, not only correlating with the concentration of MSCs but also the action between ceils and the secretory cytokine of MSCs.

关 键 词：间充质干细胞 B淋巴细胞 免疫球蛋白 免疫调节 

分 类 号：R392[医药卫生—免疫学]