乙型肝炎慢加急性肝功能衰竭患者免疫失衡状态下单核细胞的研究

Study of peripheral blood monocytes in patients with hepatitis B virus-related acute-on-chronic liver failure under immune dysfunction state

目的研究免疫失衡状态下乙型肝炎慢加急性肝功能衰竭（ACLF）患者单核细胞在凋亡和抗原提呈方面的改变。方法收集26例乙型肝炎ACLF患者、20例活动性慢性乙型肝炎（CHB）患者和18名健康对照者的外周血标本。应用流式细胞术检测单核细胞凋亡和增殖（Ki67）的改变，以及单核细胞表面抗原提呈分子人类白细胞抗原DR（HLA-DR）和B7分子（CD86）的表达水平。结果ACLF组患者单核细胞膜联蛋白V的表达（64％）明显高于CHB组（28％）和健康对照组（20％），差异均有统计学意义（χ2=11．75和27．23，P值均〈O．01），提示单核细胞凋亡增加。ACLF组单核细胞Ki67的表达（32％）低于健康对照组（61％）和CHB组（64％），差异均有统计学意义（χ2=4．71和4．83，P值均〈O．05），提示活化的单核细胞减少。ACLF组患者单核细胞HLA—DR和CD86的平均荧光强度为22．85和11．63，明显低于健康对照组和CHB组，提示单核细胞抗原提呈能力受损。ACLF组死亡患者单核细胞膜联蛋白V阳性表达（63％）明显高于存活者（46％）。结论处于免疫失衡状态下的乙型肝炎ACLF患者外周血单核细胞的凋亡增加，活化细胞数量减少，从而导致单核细胞抗原提旱能力的下降。

Objective To study apoptosis and antigen presentation changes of monocytes in HBV-related acute-on-chronic liver failure （ACLF） patients under immune dysfunction state. Methods Peripheral blood samples of 26 HBV-related ACLF patients （ACLF group）, 20 active chronic hepatitis B patients （CHB group） and 18 healthy individuals （control group） were collected. The changes of apoptosis and proliferation （Ki67） in monocytes and the expression of surface markers including human leukocyte antigen （HLA）-DR and ]37 molecules （CD86） of monocytes were analyzed by flow cytometry. Results The percentage of Annexin V expressed monocytes of ACLF group （64%） was significantly higher than that of CHB group （28%） and control group （20%）, and the difference was statistically significant （χ2= value was 11.75 and 27.23 ; both P〈0.01）, which indicated that monocytes apoptosis increased. The Ki67 expression in monocytes of ACLF group was lower than that of CHB group and control group, and the difference was statistically significant （χ2 value was 4.71 and 4. 83） both P〈2 0. 05）, which indicated that activated monocytes reduced. The mean fluorescence intensity （MFI） of HLA-DR and CD86 of monocytes in ACLF group was 22. 85 and 11.63, which was significantly lower than that of CHB group and control group, indicating the antigen presentation ability of monocytes injured. The percentage of Annexin V positive monocytes in survivals （62% ） was significantly higher than that of dead patients （46%） in ACLF group. Conclusion In HBV related ACLF patients under immune dysfunction state, the apoptosis of peripheral blood monocytes increased, and the quantity of activated cells reduced, resulting in the decline of the antigen presentation ability of monocytes.