间充质干细胞对异体B淋巴细胞增殖和凋亡的影响  

Effects of mesenchymal stem cells on proliferation and apoptosis of allogeneic B lymphocytes

作　　者：武令启[1] 白海[1] 王存帮[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：《中华医学杂志》2008年第36期2562-2565,共4页National Medical Journal of China

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（MSC）在体外对异体外周血B淋巴细胞增殖和凋亡的影响。方法从骨髓中分离、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，终浓度10μg／ml）刺激与不同比例的MSC（B：MSC 50：1、10：1、1：1）及不同浓度的MSC培养上清（12．5％、25％、50％）共培养的B淋巴细胞，72h后四甲基偶氮唑蓝（MTY）比色分析测B淋巴细胞的增殖。B淋巴细胞与MSC等比例，加或不加Anti—IgM，共培养24、48h后，流式细胞术检测B淋巴细胞的凋亡。结果10：1、1：1组B淋巴细胞A值分别为0．418±0．103、0．365±0．114，显著低于对照组的0．679±0．049（P〈0．01），1：1组显著低于50：1组（P〈0．05）；50％MSC上清组B淋巴细胞A值（0．504±0．099），同对照组相比差异有统计学意义（P〈0．05），并且显著低于12．5％组（0．669±1．023，P〈0．05）。MSC不诱导B淋巴细胞的凋亡；B淋巴细胞与MSC共培养24、48h，加或不加Anti—IgM，各组细胞凋亡率为（1．90±0．75）％、（2．33±1．01）％、（2．33±0．75）％、（1．39±0．63）％，组间比较差异无统计学意义（P〉0．05）。结论MSC及其上清抑制B淋巴细胞增殖，作用机制与MSC细胞数量和MSC分泌的细胞因子有关，但是在体外MSC不诱导B淋巴细胞的凋亡。Objective To study the influence of bone marrow mesenchymal stem cells （MSC） on the proliferation and apoptosis of allogeneic peripheral B lymphocytes in vitro. Methods MSCs were isolated from the peripheral blood of healthy volunteer and cultured. B lymphocytes were isolated from another healthy volunteer and co-cultured with the MSCs at the B ： MSC ratios of 50 ： 1,10 ： 1, and 1 ： 1 or with different concentrations of MSC supernatant （ 12. 5% , 25% , and 50% ） for 72 h, in the presence of antihuman IgM immunoglobubin goat antibodies （ Anti-IgM ） at a final concenrtration of 10 μg/mlL. The proliferation of B lymphocytes was analyzed with MTr assay. B lymphocytes and MSC were co-cultured at the ratio of 1 ： 1 for 24 h or 48 h, with or without addition of Anti-IgM. Flow cytometric was used to detect the apoptosis of B lymphocytes. Results The A value of B lymphocytes co-cultured with MSCs at different ratios were 0.521 ±0.093, 0.418 ±0. 103, and 0.365 ±0. 114 respectively. The A values of Group10：1 and Ggroupl： 1 were both significantly lower than that of the control group （0. 679 ±0. 049, both P 〈0. 01 ） ,and the A value of Ggroupl ： 1 was significantly lower than that of Group10：1 （ P 〈 0. 05 ）. The A value of B lymphocytes co-cultured with 50% MSC supernatant was 0. 504 ± 0. 099, significantly lower than those of the control group and Group 12. 5% （both P 〈0. 05）. MSCs didn＇t induce apoptosis of B lymphocytes. The apoptosis rates of B lymphocytes co-cultured with MSCs for 24 h or 48 h, in presence or absence of Anti-IgM were 1.90% ±0. 75%, 2. 33% ± 1.01%, 2. 33%± 0. 75%, and 1.39% ± 0. 63% respectively, without significant difference between any 2 of the four groups. Conclusion MSC and its supernatant inhibit B lymphocyte proliferation with the mechanism correlated with the MSC concentration and the MSC-secreted cytokine, but MSCs does not induce B lymphocytes apoptosis in vitro.

关 键 词：间充质干细胞 B淋巴细胞 免疫调节 凋亡 

分 类 号：R686[医药卫生—骨科学]