乙型肝炎病毒感染对造血干细胞活性的影响  

Impact of hepatitis B virus infection on the activity of hematopoietic stem cell

作　　者：石雁梅[1] 兰英华[1] 单蕾[1] 蔡华峰[1] 孙静霞[1] 王永晨[1] 周晋[1] 李用国[1] 

机构地区：[1]哈尔滨医科大学附属第一医院感染科,哈尔滨150001

出　　处：《中华传染病杂志》2008年第4期197-201,共5页Chinese Journal of Infectious Diseases

基　　金：基金项目：国家自然科学基金资助项目（30571638）

摘　　要：目的了解HBV感染对脐血来源的造血干细胞活性的影响。方法将分离纯化的健康脐血CD34^＋细胞，在含有干细胞生长因子（SCF）、酪氨酸激酶受体家族Ⅲ配体（FL）、促血小板生成素（TPO）、IL-3和10％FBS的IMDM培养基中扩增，同时加入高拷贝HBV，观察干细胞扩增与病毒复制规律；干细胞扩增后，在巨细胞集落刺激因子（GM-CSF）和IL-4作用下诱生树突状细胞，对干细胞及树突细胞进行形态学观察，并检测其表面分子的表达。两组间比较采用独立t检验，多组间比较采用方差分析。结果感染HBV的造血干细胞自然增殖能力明显低于正常干细胞（P〈0．01）；加入细胞因子后细胞增殖增加（P〈0．01），细胞内HBVDNA复制也增加（P〈0．01），但其干细胞增殖仍低于正常干细胞加细胞因子组（P〈0．05）。电子显微镜观察发现HBV感染干细胞后胞质内出现Dane颗粒；HBV感染的干细胞经诱导为树突状细胞后，其免疫表型CD80、CD86、CD1a、HLA-DR的表达均低于未感染组（P〈0．01）。结论HBV可以感染CD34^＋造血干细胞，并随干细胞增殖而复制增加，HBV不仅抑制造血干细胞增殖，而且下调干细胞分化来源的树突状细胞免疫表型的表达。Objective To study the impact of hepatitis B virus （HBV） infection on the activity of cord hematopoietic stem cells. Methods CD34^＋ cells were isolated from healthy human cord blood by mini MACS. Cells were cultured in IMDM complete culture medium containing stem cell factor （SCF）, fms-like tyrosine kinase 3 ligand （FL）, thrombopoietin （TPO）, interleukin-3 （IL-3） and 10% fetal bovine serum. High copies HBV were added to the culture system. The proliferation of stem cells and virus replication were observed. Following the proliferation, dendritic cells （DCs） were induced by adding granulocyte-macrophage colony-stimulating factor and IL-4. Morphous of stem cells and DCs were observed by microscope and the cell surface molecules were detected. Results The proliferation of stem cells infected with HBV was significantly lower than that of healthy stem cells （P〈0.01）, and enhanced after adding cytokines （P〈0.01）. At the same time, HBV replication was increased after adding cytokines in the culture system （P〈0.01）, but the proliferation was still lower than that of healthy stem cells with cytokines in the culture medium （P〈0.05）. Dane particles were found in the cytoplasma of stem cells infected with HBV by electron microscope. The expression of CD80, CD86,CD1a and HLA-DR on DCs derived from HBV infected stem cells were all lower than those on DCs from non-infected stem cells （P〈0.01）. Conclusions HBV could infect CD34^＋ stem cell and the proliferation of the stem cell could enhance the virus replication. HBV could not only inhibit the proliferation of stem cells, but also down-regulate the immuno-phenotype expression of DCs derived from CD34^＋ stem cells.

关 键 词：造血干细胞 肝炎病毒 乙型 树突细胞 免疫表型分型 

分 类 号：R512.62[医药卫生—内科学]