慢性乙型重型肝炎患者肝组织中浸润淋巴细胞中免疫活性细胞的研究

目的 明确慢性乙型重型肝炎（简称慢重肝）患者肝内浸润的淋巴细胞（LILs）及外周血中免疫活性细胞的频率,并对其进行对比分析,以明确它们的异同以及在慢重肝发病机理中的作用。方法 通过毛玻璃研磨加自然沉降肝细胞的方法,对11例慢重肝接受肝移植患者的LILs进行提取、分离。通过流式细胞仪测定慢重肝患者LILs及外周血中免疫活性细胞的频率并行对比分析,同时与正常人LILs及外周血中免疫活性细胞的频率进行对比分析。结果 （1）慢重肝患者LILs中CD4^＋T淋巴细胞及B细胞的频率分别为17％±6％及3．0％±1．0％,明显低于外周血中各自的频率,分别为32％±8％及21．4％±12．2％（均P〈0．01）,而CD8^＋T淋巴细胞、NK及NKT细胞的频率分别为38％±13％、34％±18％及10％±4％,明显高于外周血中各自的频率,分别为26％±6％、15％±9％及6％±4％（均P〈0．05）;（2）与正常对照相比,LILs中CD3^＋及CD4^＋T淋巴细胞明显要高（P〈0、05或P〈0、01）,而NK细胞及NKT细胞要低些;（3）与正常对照相比,慢重肝患者uIJs与外周血淋巴细胞之比,CD3^＋、CD4^＋、CD8^＋T淋巴细胞有更高的比值（CD4^＋T淋巴细胞P〈0、01）,B细胞及NKT细胞有更低的比值（P〈0．01或P〈0、05）。结论 肝脏内大量免疫活性细胞的浸润,尤其是CD4^＋、CD8^＋T淋巴细胞、NK细胞的大量浸润对于慢重肝的发病可能起重要的作用。

Immunologically-competent cells in liver infiltrating lymphocytes in patients with chronic severe hepatitis B

OBJECTIVE: To investigate the frequencies of immunologically competent cells (ICCs) in the liver-infiltrating lymphocytes (LILs) and peripheral blood and their possible role in pathogenesis in patients with chronic severe hepatitis B (CSHB). METHODS: LILs were isolated from the liver tissue samples from 11 CSHB patients and 5 normal controls (NCs) by the method of combined grinding with semi-frosted microscopic slides and sedimentation of hepatic cells. The frequency of isolated ICCs, including CD3(+), CD4(+), and CD8(+) T-cells, NK cells, NKT cells, and B cells was examined and compared with that of the circulating ICCs in the CSHB patients. Comparison was conducted between the CSHB patients and the controls. RESULTS: (1) In the CSHB patients, the frequencies of CD4(+) T cells and B cells in LILs were 17% +/- 6% and 3.0% +/- 1.0% respectively, both significantly lower than those in the circulating blood (32% +/- 8% and 21.4% +/- 12.2% respectively, both P < 0.01); however, the frequencies of CD8(+) T cells, NK cells, and NKT cells in LILs were 38% +/- 13%, 34% +/- 18%, and 10% +/- 4% respectively, all significantly lower than those in the circulating blood (26% +/- 6%, 15% +/- 9%, and 6% +/- 4%, all P < 0.05). (2) The frequencies of infiltrating CD3(+) T cells and CD4(+) T cells of the CSHB patients were both significantly higher than those of the NCs (P = 0.042 and P = 0.001); and the frequency of infiltrating CD8(+) T cells of the CSHB patients was higher than that of the NCs, and the and the frequencies of infiltrating NK cells and NKT cells in LILs were lower than those of the NCs, however, not significantly. (3) Compared with the liver tissues from the NCs, the liver tissues from the CSHB patients exhibited a significantly higher ratio of liver-infiltrating CD4(+) T cells to peripheral blood CD4(+) T-cell subsets (P = 0.001), and significantly lower ratios of liver-infiltrating NKT cells and B cells to the peripheral blood NKT-cells and B cells (P = 0.029 and P = 0.001 respectively). CONCLUSION: The abundant infiltrating immune active cells, especially the CD4(+) T cells, CD8(+) T cells, and NK cells, may be the causal factors that drive the progressive development of CSHB.