人脐带间充质干细胞对难治性免疫性血小板减少症患者B淋巴细胞分化及分泌功能的影响  

Impacts of hUC-MSCs on the Secretion and Immune Function of B Lymphocytes in Refractory Immune Thrombocytopenia Patients

作　　者：曾玉晓[1,2] 徐玲珑[1] 吴迪炯[1] 叶宝东[1] 高雁婷[1] 刘文宾[1] 周郁鸿[1] 

机构地区：[1]浙江中医药大学第一临床医学院血液科,杭州310006 [2]浙江省丽水市人民医院血液科,丽水323000

出　　处：《中华临床免疫和变态反应杂志》2016年第3期207-212,共6页Chinese Journal of Allergy & Clinical Immunology

基　　金：浙江省医药卫生科学研究项目(2010KYB077);国家自然科学基金(81373634);浙江省中医药优秀青年人才基金计划(2015ZQ012)

摘　　要：目的探讨人脐带间充质干细胞(human umbilical cord mesenchymal stem cells,h UC-MSCs)对难治性免疫性血小板减少症(refractory immune thrombocytopenia,r ITP)患者B淋巴细胞分化及分泌功能的影响。方法酶消化法提取健康胎儿h UC-MSCs;密度梯度离心法分离2012年6月至2013年10月浙江省中医院26例r ITP患者和6名健康供者的外周血单个核细胞;美洲商陆(Pokeweed,PWM)刺激外周血B细胞分化;酶联免疫吸附测定法(enzyme-linked immuno sorbent assay,ELISA)检测r ITP患者和健康供者外周血单个核细胞和h UC-MSCs共培养时IgG、IgM的分泌量;En Vision免疫组化法测B细胞诱导的成熟蛋白-1(B lymphocyte-induced maturation protein1,Blimp-1)表达。结果 (1)流式细胞技术测健康胎儿脐带提取的h UC-MSCs表面CD44表达为99.8%。(2)PWM浓度为30 mg/L,作用4 d时r ITP患者组IgG、IgM分泌量分别为(7.39±1.42)和(5.44±0.17)μg/L,明显高于健康供者组(P=0.014、0.039)。(3)h UC-MSCs和r ITP患者外周血单个核细胞以2∶10共培养时IgG、IgM分泌量分别为(4.98±1.63)μg/L和(3.78±0.82)μg/L,明显低于无h UC-MSCs时IgG、IgM分泌量(P=0.005、0.003),且和健康供者组差异无统计学意义(P=0.266、0.543);此比例下rITP患者组Blimp-1蛋白表达率为33.51%,较无h UC-MSCs时明显下降(P=0.026),与健康供者组比差异无统计学意义(P=0.076)。结论 hUC-MSCs和r ITP患者外周血单个核细胞以2∶10比例共培养,可明显减少rITP患者IgM、IgG的分泌量,抑制Blimp-1蛋白的表达,提示hUC-MSCs对r ITP患者B细胞分化为浆细胞分泌抗体有一定的抑制作用,为临床hUC-MSCs治疗rITP提供一定的实验依据。Objective To explore the impacts of hUC-MSCs on the secretion and immune function of B lymphocytes in refractory idiopathic thrombocytopenic purpura patients. Methods Enzyme digestion was used to extract healthy fetus umbilical cord mesenchymal stem cells; The 26 rlTP patients who were in the Zhejiang Provincial Hospital of TCM from June 2012 to October 2013 and six healthy donors were included into this study, The peripheral blood mononuclear cells （PBMCs） of them were extracted by density gradient centrifugation; Pokeweed （PWM） was used to stimulate B lymphocyte transferring to plasmocytes for secretion of IgG and IgM. The concentration of IgG and IgM of the rITP patients weretested by enzyme-linked immuno sorbent assay （ELISA） when their which PBMCs were co-cultured with hUC-MSCs, so did for healthy donors. The epression of Blimp-1 was measured by the EnVision immunohistochemistry. Results （1） The cells extracted from healthy fetus umbilical cord tested by flow cytometry labeled for CD44 was 99. 8%. （2） The antibody IgG and IgM secreted from the rITP patients was （ 7.39 ± 1.42 ） μg/L and （ 5.44 ± 0. 17 ） μg/L respectively, that was significantly increased than healthy doners, when the concentration of PWM was 30 mg/L and the cells were stimulated for 4 days （P =0. 014, 0. 039） . （3） When the hUC-MSCs and the PBMCs was at a 2：10 ratio, the IgG and IgM concentration of rlTP patients was （4. 98 ± 1.63） μg/L and （3.78 ±0. 82） μg/L respectively, which was declined sharply than non-hUC-MSCs＇ （P = 0. 005, 0. 003 ）, and not much difference with healthy donors （P =0. 848）. The rate of Blimp-1 protein of rITP patients was 33.51%, that apparently declined at the ratio 2：10 of hUC-MSCs/ peripheral blood mononuclear ceils （P = 0. 026）, but no significant difference when compared to healthy donors （P = 0. 076）. Conclusions The hUC-MSCs co-cultured with rITP patiens＇ peripheral blood mononuclear cells at 2：10 ratio could decline IgG and IgM secreti

关 键 词：血小板减少 间质干细胞 B淋巴细胞 

分 类 号：R558.2[医药卫生—血液循环系统疾病]