慢性乙型肝炎病毒携带者外周血单个核细胞凋亡及细胞因子检测的意义

Detection of peripheral blood mononuclear cell and cytokine of supernatant induced by PHA in chronic hepatitis B virus carriers

目的探讨外周血单个核细胞凋亡（PBMC）和Th1／Th2型细胞因子表达在乙型肝炎病毒慢性化中的作用。方法分离21例健康人、45例慢性乙型肝炎病毒携带者PBMC，在植物血凝素（PHA-P）刺激培养72h后，以酶联免疫吸附测定法检测细胞培养上清液中的γ-干扰素（IFN-γ）浓度，流式细胞仪以膜联蛋白V-异硫氰酸荧光素／碘化丙锭（AnnexinV-FITC／PI）法检测PBMC凋亡率。结果慢性乙型肝炎病毒携带者PBMC凋亡率明显高于对照组（P〈0．01），培养上清液IFN-γ水平低于对照组（P〈0．01），且与PBMC凋亡率呈负相关（r=-0．647，P〈0.01）；IL4水平高于正常对照组（P〈0.01），且与PBMC凋亡率呈正相关（r=0．598，P〈0．01）。结论严重活化诱导的淋巴细胞凋亡（AICD）和Th1／Th2失衡在乙型肝炎病毒慢性化中起一定的作用。

Objective To discuss the significance of peripheral blood mononuclear cell（PBMC） apoptosis and Thl/Th2 cytokine in chronic hepatitis B virus carriers. Methods PBMC was extracted from healthy people （21 subjects） and chronic hepatitis B virus carriers （45 cases）. PBMC apoptosis was stained with Annexin V-FITC./PI and assessed by flow cytometry after PBMC was cultured for 72 hours in vitro with phytohemagglutinin P （PHA-P）. The level of interferon-γ/（IFN-γ） and IL-4 from the upper culture fluid were measured by speclfic-enzyme linked immunosorbent assay （ELISA）. Results The apoptosis of PBMC was higher in chronic hepatitis B virus carriers than in control group（P 〈 0.01 ）. The level of IFN-γ was lower in chronic hepatitis B virus carriers than that in control group（ P 〈 0.01 ） and negatively correlated with apoptosis of PBMC（ r = -0. 647 ,P 〈 0.01 ）. The level of IL-4 was higher in chronic hepatitis B virus carriers than that in control group（ P 〈 0.01 ） and positively correlated with apoptosis of PBMC （ r = 0. 598, P 〈 0.01 ）. Conclusion More severity activation-induced cell death （AICD） and imbalance of Thl/Th2 had negative effect on chronic hepatitis B virus carriers.