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目的 研究慢性乙型肝炎患者外周血T淋巴细胞 (PBL T)激活诱导细胞死亡 (AICD)的发生情况 ,探讨AICD在慢性乙型肝炎发病机制中的作用。方法 采用免疫磁珠技术负性分离 14例慢性乙型肝炎患者和 9例正常人PBL T ,应用抗CD3单克隆抗体、佛波醇 (PMA)及离子霉素 (iono mycin)体外诱导细胞活化 ;采用脱氧核苷酸转移酶介导的dUTP切口末端标记 (TUNEL)染色观察凋亡情况并用流式细胞仪检测凋亡率。结果 慢性乙型肝炎患者诱导组PBL T凋亡率为 (16.73±0 .99) % ,明显高于未诱导组 (9.74± 1.14 ) % ,P <0 .0 1。无论是否诱导 ,患者PBL T的凋亡率均高于健康人。培养上清液中IFN γ浓度与T细胞凋亡率成负相关 (P <0 .0 1) ,而各组IL 10的浓度差异无显著性 (P >0 .0 5 )。结论 慢性乙型肝炎患者PBL T存在AICD现象 ,并且可能是造成HBV持续感染的重要原因
Objective To study the occurrence of peripheral blood mononuclear cells (PBMC) T-lymphocyte-induced death (AICD) in patients with chronic hepatitis B and to explore the role of AICD in the pathogenesis of chronic hepatitis B. Methods PBL T was negatively isolated from 14 patients with chronic hepatitis B and 9 normal individuals by immunomagnetic beads technique. The cells were induced by anti - CD3 monoclonal antibody, phorbol myristate (PMA) and ionomycin in vitro. Apoptosis was observed by dUTP nick end labeling (TUNEL) staining and the apoptosis rate was detected by flow cytometry. Results The apoptosis rate of PBL T in induced group was (16.73 ± 0.99)%, which was significantly higher than that in non-induced group (9.74 ± 1.14)%, P <0.01. Patients with or without induction of PBL T apoptosis rate was higher than healthy people. IFNγ concentration in culture supernatant was negatively correlated with T cell apoptosis rate (P <0.01), while the concentration of IL 10 in each group had no significant difference (P> 0.05). Conclusions There is AICD phenomenon in PBL T of patients with chronic hepatitis B, and it may be the important reason of persistent HBV infection