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.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
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）
