摘要
目的探讨脂肪来源的间充质干细胞(ADSCs)对原发免疫性血小板减少症(ITP)患者CD4^(+)T细胞分化及Th1/Th2因子表达的影响,并探讨相关机制。方法分离培养及鉴定健康人来源的ADSCs,观察干细胞形态及增殖分化情况,选取增殖稳定的ADSCs;选取华中科技大学协和深圳医院血液科ITP患者及招募健康对照者各6例,分离两组外周血CD4^(+)T细胞,并与ADSCs共培养。观察ADSCs对CD4^(+)T分化的影响,Th1/Th2细胞亚群比及特异性转录因子表达变化,并检测Th1/Th2因子表达水平。结果(1)收集的ADSCs生长状态良好,表达抗原CD29、CD44、CD105,而低表达或不表达CD14、CD31、CD34、CD45、CD71、CD106及CD117,经鉴定证明其具有多向分化能力。(2)ITP患者Th1/Th2亚群在CD4^(+)T细胞中所占的比例较健康对照组高(P<0.05);ADSCs可直接抑制ITP患者CD4^(+)T细胞向Th1分化,促进向Th2分化,降低Th1/Th2比值(均P<0.05),而对健康对照者CD4^(+)T细胞分化影响不明显(P>0.05)。(3)ITP患者CD4^(+)T细胞与ADSCs共培养前后比较发现,Th1特异性转录因子GATA-3表达减少,Th2特异性转录因子T-bet表达升高,同时Th1因子IL-2、IFN-γ的表达下降,而Th2因子IL-4、IL-10等表达上升,差异均具有统计学意义(均P<0.05)。结论ITP患者存在CD4^(+)T细胞亚群免疫失衡,主要表现为Th1偏移,ADSCs可能通过对特异性转录因子GATA-3/T-bet的转录调控,抑制Th1细胞分化、促进Th2细胞分化,调节Th1/Th2细胞因子表达,继而改善免疫失衡。
Objective To explore the effect of adipose-derived mesenchymal stem cells(ADSCs)on CD4^(+)T cell differentiation and Th1/Th2 expression in idiopathic thrombocytopenic purpura(ITP)patients and the related mechanism.Methods After isolating,culturing and identifying ADSCs from healthy human,the morphology,proliferation and differentiation of stem cells were observed.ADSCs with stable proliferation were selected.In all,6 ITP patients in the hematology department of our hospital and 6 healthy controls were recruited,and CD4^(+)T cells were isolated from their peripheral blood samples.CD4^(+)T cells were collected and co-cultured with ADSCs to observe the effect of ADSCs on CD4^(+)T cell differentiation.Ratio of Th1/Th2 cell subsets,expression of specific transcription factors and expression level of Th1/Th2 factors were detected.Results(1)The collected ADSCs grew well.CD29,CD44,and CD105 were expressed in the collected ADSCs,while CD14,CD31,CD34,CD45,CD71,CD106,and CD117 were low or not expressed.And they were identified to have the ability of multidirectional differentiation.(2)The ratio of Th1/Th2 subsets in ITP patients was higher than that in the healthy controls(P<0.05).ADSCs directly inhibited the differentiation of CD4^(+)T cell into Th1,but yet promoted the differentiation of CD4^(+)T cell into Th2,therefore reduced Th1/Th2 ratio(all P<0.05).However,the effect on CD4^(+)T cell differentiation in healthy subjects was not significant(P>0.05).(3)After co-culture with ADSCs,the expression of Th1-specific transcription factor GATA-3 was decreased,while the Th2-specific transcription factor T-bet expression was increased.The expression levels of Th1 factors IL-2 and IFN-γdecreased,while the expression levels of Th2 factors IL-4 and IL-10 increased.The differences were statistically significant(all P<0.05).ConclusionThe immune imbalance of CD4^(+)T cell subsets in ITP patients is mainly manifested as Th1 deviation.Through the transcriptional regulation of specific transcription factor GATA-3/T-bet,ADSCs would inhibit Th1 cell differentiation and promote Th2 cell differentiation,then regulate Th1/Th2 cytokine expression,therefore reverse immune imbalance.
作者
肖建红
张阳春
刘泽林
Xiao Jianhong;Zhang Yangchun;Liu Zelin(Department of Hematology,Union Shenzhen Hospital,Huazhong University of Science and Technology,Shenzhen518052,China;Department of Hematology,The First Affiliated Hospital of South China University,Shenzhen518101,China;Department of Orthopedics,People’s Hospital of Shenzhen Baoan District,Hengyang421001,China)
出处
《华中科技大学学报（医学版）》
CAS
CSCD
北大核心
2022年第3期375-382,共8页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
基金
国家自然科学基金资助项目(No.81500091)
湖南省自然科学基金青年基金项目(No.2018JJ3472)
广东省医学科研基金资助项目(No.B2020020)。
关键词
脂肪间充质干细胞
血小板减少症
CD4
T细胞
免疫调节
adipose-derived mesenchymal stem cells
idiopathic thrombocytopenic purpura
CD4+T cell
immunoregulation
