乙型肝炎患者骨髓造血干细胞来源的树突状细胞表型分析

目的 检测乙型肝炎患者骨髓造血干细胞（HSC）诱导的树突状细胞（DC）的表面分子的表达,评价其相关因素及临床意义.方法 收集慢性乙型肝炎患者的骨髓液9例,健康者7例后用磁珠分离仪分离纯化骨髓液CD34＋细胞,在含有干细胞生长因子（SCF）、酪氨酸激酶受体家族Ⅲ的配体（FLT3）、促血小板生成素（TPO）、IL-3和10%FBS的IMDM培养基中孵育并进行扩增,在干细胞扩增基础上,在GM-CSF和IL-4作用下诱生DC,通过流式细胞仪分析其表面分子的表达并对DC进行形态学观察.结果 乙型肝炎患者HSC经诱导为DC后,其免疫表型CD80、CD86、CD1a的表达均低于正常对照组,差异有统计学意义：CD1a（t=3.94,P＜0.05）,CD80（t=7.08,P＜0.01）,CD86（t=3.65,P＜0.05）,而HLA-DR表达与正常组比较差异无统计学意义（t=0.34,P＞0.05）.结论 慢性乙型肝炎患者HSC来源的部分DC的表面分子表达低下可能与HBV感染HSC后出现病态的免疫细胞分化有关.

The phenotypes of dentritic cells derived from marrow haematopoietic stem cells of patients with hepatitis B

Objective To determine the expression of surface molecules of dentritic cells (DCs) derived from marrow haematopoietic stem cell (HSC) of patients with hepatitis B and its clinical significance.Methods The marrows were collected from 9 patients with hepatitis B and 7 healthy donors. The CD34 + cells were isolated and purified by mini-MACS ,then seeded in 12-well plates and cultured in IMDM complete culture medium with 10% FCS and cytokines (SCF,FL3, TPO, IL-3 ). Following the proliferation, DCs were induced by adding GM-CSF,IL-4,TNF for extra 7 days, and the expression of surface molecules on DCs was examined by flow cytometry and the morpholog of stem cells and DCs were observed by microscopy. Results The expression of CD80, CD86, CD1a on DCs derived from haematopoietic stem cells of patients with hepatitis B was in low level compared with that of DCs from healthy donor: CD1a: t = 3.94, P< 0.05; CD80: t = 7. 08, P < 0. 01;CD86:t = 3. 65 ,P <0.05. But the expression of HLA-DR in both gnoups was not significant different ( t =0. 34,P > 0. 05 ) Conclusion The expression of surface molecules on DCs derived from marrow haematopoietic stem cell of patients with hepatitis B decreaded compared with healthy control, which may be related to unhealthy dif ferentiation of haematopoietic stem cells with infection of HBV.