CAJ | 학술논문

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HBV DNA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.
목적 료해HBV감염대제혈래원적조혈간세포활성적영향.방법 장분리순화적건강제혈CD34+세포,재함유간세포생장인자(SCF)、락안산격매수체가족Ⅲ배체(FL)、촉혈소판생성소(TPO)、IL-3화10%FBS적IMDM배양기중확증,동시가입고고패HBV,관찰간세포확증여병독복제규률;간세포확증후,재거세포집락자격인자(GM-CSF)화IL-4작용하유생수돌상세포,대간세포급수돌세포진행형태학관찰,병검측기표면분자적표체.량조간비교채용독립t검험,다조간비교채용방차분석.결과 감염HBV적조혈간세포자연증식능력명현저우정상간세포(P<0.01);가입세포인자후세포증식증가(P<0.01),세포내HBV DNA복제야증가(P<0.01),단기간세포증식잉저우정상간세포가세포인자조(P<0.05).전자현미경관찰발현HBV감염간세포후포질내출현Dane과립;HBV감염적간세포경유도위수돌상세포후,기면역표형CD80、CD86、CD1a、HLA-DR적표체균저우미감염조(P<0.01).결론 HBV가이감염CD34+조혈간세포,병수간세포증식이복제증가,HBV불부억제조혈간세포증식,이차하조간세포분화래원적수돌상세포면역표형적표체.
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 miniMACS. 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 ceils 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.