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目的探讨外周血及肝组织T淋巴细胞亚群、自然杀伤细胞(natural killer cells,NK细胞)在慢性乙型肝炎不同临床转归中的变化及其意义。方法慢性乙型肝炎(chronic hepatitis B,CHB)患者34例;HBV相关肝衰竭患者54例,其中,慢性肝功能衰竭(chronicliverfailure,CLF)31例,慢加急性肝功能衰竭(acute-on-chronic liver failure,ACLF)23例。10例CHB患者经肝脏活检、5例ACLF患者行肝脏移植获得肝组织标本。流式细胞仪检测外周血CD4+CD25+FOXP3+调节性T细胞(regulatory T cells,Treg细胞)、CD3+、CD4+、CD8+T淋巴细胞亚群,免疫组织化学观察肝组织中CD8+T细胞、NK细胞频率。结果 ACLF患者外周血Treg细胞频率(4.79%±2.58%)明显高于CLF患者(3.24%±2.22%)及CHB患者(3.46±1.78%,P<0.05);CLF患者Treg细胞频率低于CHB患者,但无显著性差异。动态观察发现,CLF患者Treg细胞频率持续低水平;而ACLF患者Treg细胞频率先升高,随着肝功能恢复,Treg细胞频率下降。CLF患者外周血CD3+、CD4+、CD8+T淋巴细胞计数分别为939.2±466.2、544.1±300.9、391.8±240.6,明显低于CHB患者(1567.4±556.6、864.2±289.4、563.3±315.1)及ACLF患者(1596.1±1151.6、923.1±701.5、576.8±392.2,P<0.01)。ACLF患者外周血T淋巴细胞亚群计数与CHB患者比较无显著性差异。免疫组化显示,ACLF患者肝组织CD8+T细胞、NK细胞浸润较CHB患者明显增加。结论 CLF患者存在严重的T淋巴细胞“耗竭”。外周血Treg细胞在ACLF发生机制中可能具有重要的免疫调节作用。肝组织CD8+、NK细胞浸润增加与慢性乙型肝炎肝脏病理恶化有关。
Objective To investigate the changes and clinical significance of T lymphocyte subsets and natural killer cells (NK cells) in peripheral blood and liver tissues in different clinical outcomes of chronic hepatitis B patients. Methods Thirty-four patients with chronic hepatitis B (CHB) and 54 patients with HBV-related liver failure (COPD), including 31 patients with chronic liver failure (CLF) and 30 patients with acute-on-chronic Liver failure, ACLF) 23 cases. Liver biopsies were obtained from 10 CHB patients and 5 ACLF patients underwent liver transplantation. Flow cytometry was used to detect the subsets of CD4 + CD25 + FOXP3 + regulatory T cells, CD3 +, CD4 + and CD8 + T lymphocytes in peripheral blood. Immunohistochemistry was used to observe the expression of CD8 + T cells, NK Cell frequency. Results The frequency of Treg cells in peripheral blood of ACLF patients (4.79% ± 2.58%) was significantly higher than that of CLF patients (3.24% ± 2.22%) and CHB patients (3.46 ± 1.78%, P <0.05) , But no significant difference. Dynamic observation found that the frequency of Treg cells in CLF patients continued to be low; however, the frequency of Treg cells in ACLF patients first increased, and the frequency of Treg cells decreased with the recovery of liver function. The counts of CD3 +, CD4 + and CD8 + T lymphocytes in peripheral blood of CLF patients were 939.2 ± 466.2, 544.1 ± 300.9 and 391.8 ± 240.6, respectively, which were significantly lower than those of patients with CHB (1567.4 ± 556.6, 864.2 ± 289.4, 563.3 ± 315.1) and ACLF 1596.1 ± 1151.6, 923.1 ± 701.5, 576.8 ± 392.2, P <0.01). ACLF patients with peripheral blood T lymphocyte subsets count and CHB patients no significant difference. Immunohistochemistry showed that ACLF patients with liver tissue CD8 + T cells, NK cells infiltration significantly increased in patients with CHB. Conclusions Serious T lymphocyte “depletion” exists in CLF patients. Peripheral blood Treg cells may play an important immunoregulatory role in the pathogenesis of ACLF. The increase of CD8 + and NK cell infiltration in liver tissue is related to the pathological deterioration of liver in chronic hepatitis B patients.