肝细胞癌患者免疫功能的研究

The study on cell immunologic function of hepatocellular carcinoma

目的探讨肝细胞癌免疫功能的临床意义。方法对54例HCC患者、64例慢性乙型肝炎患者和23位正常献血员采用双抗体夹心法,一抗为生物素标记的抗IFN-γ、TNF-α、IL-2、IL-4、IL-10、IL-12P70抗体,根据样本的OD值及曲线的斜率计算出细胞因子的含量。采用免疫组织化学方法检测外周血CD3+、CD4+和CD8+T淋巴细胞的百分率。银染法检测T淋巴细胞核仁形成区嗜银蛋白,流式细胞仪测定肝细胞癌患者外周血黏附单个核细胞CD83及HLA-DR表达。结果 HCC患者血清IFN-γ含量为(4.92±2.15)pg/mL,低于健康献血员(19.16±2.34)pg/mL和慢性乙型肝炎患者(11.42±6.86)pg/mL(P<0.05)。HCC患者IL-2、IL-12P70含量降低,而TNF-α、TGF-β1、IL-4、IL-10含量增高。根据肝癌体积大小对患者分组比较血清IFN-γ水平,随着肿瘤体增大,血清IFN-γ水平下降,调节IFN-γ分泌的IL-12P70水平与IFN-γ的变化趋势一致,但HCC患者血液中都存在IFN-γ。根据甲胎蛋白的水平将未经治疗的HCC患者分成2组,结果提示甲胎蛋白含量高时血清IFN-γ水平下降,调节IFN-γ分泌的IL-12P70水平与IFN-γ的变化趋势一致。与慢性乙型肝炎患者和健康献血员相比,HCC患者CD4+T淋巴细胞、CD4+/CD8+比值降低,反映T淋巴细胞增殖活性的AgNORs降低。黏附细胞悬液CD83和HLA-DR表达降低。结论 HCC患者的免疫功能低下,与肿瘤细胞的免疫逃逸有关。

Objective To investigate the clinical significance of immunologic function of hepatocellular carcinoma（HCC）.Methods Periperal blood of 54 cases of HCC patients,64 cases of chronic hepatitis B patients and 23 cases of healthy blood donors were collected.Double antibody sandwich ELISA method was used.Primary antibody was biotin-labeled antibody which resisted IFN-γ,TNF-α,IL-2,IL-4,IL-10 and IL-12P70.The cytokine content was calculated according to the OD value of the sample and the slope of the curve.The percentage of the lymphocytes of CD3＋,CD4＋and CD8＋T in the peripheral blood were measured by immunohistochemical method.Argentation was adopted to measure the amount of argyrophilia protein in the nucleolus organizer of the T lymphocytes.The flow cytometry was employed to detect the expression of mononuclear cells of CD83 and HLA-DR adherent to culture dish of the HCC patients＇ peripheral blood,respectively.Results The content of IFN-γ in the serum of HCC patients was（4.92±2.15） pg/mL,lower than that in the healthy blood donors（19.16 pg/mL±2.34 pg/mL） and chronic hepatitis B patients（11.42 pg/mL±6.86 pg/mL）（P0.05）,and contents of IL-2 and IL-12P70 were also lower,but contents of TNF-α,TGF-β1,IL4 and IL-10 were higher.The HCC patients were divided into different groups according to the size of the tumors.The level of serum IFN-γ decreased with the increament of the size of tumors.The untreated HCC patients were divided into 2 groups according to the level of α-fetoprotein.The comparative study showed that the IFN-γ level declined as the α-fetoprotein rised,but the level of IL-12P70 which regulates the level of IFN-γ secretion had the same tendency with the level of IFN-γ.The percentn age of CD4＋ peripheral blood T lymphocytes,CD83＋and HLA-DR＋ in HCC patients were lower than those in the chronic hepatitis B patients and the blood donors.AgNORs which represented the proliferative activity of the T lymphocytes decreased in patients with HCC.Conclusion Immunologic function of HCC patients decreased,and it was related with immunologic escape of tumer cells.