【作者】 于海华；

【导师】 汤钊猷； 邱双健； 王鲁； 刘彬彬；

【作者基本信息】 复旦大学 ， 肝肿瘤外科， 2008， 博士

【摘要】 目的:探讨肝癌患者系统性细胞免疫状态及临床意义;分析肝癌切除对细胞免疫的影响。方法:利用流式细胞术检测肝癌患者和健康对照组外周血T淋巴细胞亚群的频数表达;并分析肝癌切除前后外周血淋巴细胞亚群频数表达的变化;利用酶联免疫吸附实验检测肝癌患者和健康对照组外周血细胞因子的表达。同时分析肝癌T淋巴细胞亚群和细胞因子表达与肝癌生物学特征的关系。结果:与健康组相比,肝癌患者外周血T淋巴细胞亚群频数分布异常;T淋巴细胞分布和肝癌生物学及TNM分期相关。CD4+/CD8+比值与CD3+T细胞比例呈明显负相关;CD4+CD25+T细胞频数表达与CD4+/CD8+比值亦呈明显负相关,与CD3+CD45R0+T细胞表达呈明显正相关。Foxp3+T细胞可能下调了CD8+GranzymeB+T细胞的表达。用CD4+CD25+Foxp3+T细胞的频数表达来反映调节性T细胞的表达状态较CD4+CD25+T细胞更为合理和科学。肝癌患者外周血Th1/Th2型细胞因子表达失衡;细胞因子表达谱和肝癌生物学及TNM分期相关。肝癌外周IL—10表达和IFN—γ呈正相关,提示二者在作用上可能存在协同性,一定条件下IL—10可能发挥出免疫正调节作用。肝癌切除可引起暂时性的免疫抑制加重;肝癌切除通过消减肿瘤负荷减少了肿瘤相关性调节性T细胞的表达。肝癌切除术后1月左右机体细胞免疫功能有改善倾向。结论:肝癌患者细胞免疫处于抑制状态,T淋巴细胞亚群表达的检测可作为评价肝癌细胞免疫状态的指标;肝细胞肝癌外周血淋巴细胞转录因子Foxp3表达的异常增加可能在肝癌细胞免疫抑制中发挥着重要作用。肝癌患者细胞免疫状态和肝癌侵袭及演进相关;异常增加的外周调节性T细胞在肝癌较早期阶段发挥调节作用。肝癌切除可能有助于抗肿瘤免疫的恢复;肝癌患者围手术期很可作为免疫干预的重要窗口期;调节性T细胞可作为免疫干预的靶点。更多还原

【Abstract】 Objectives:The present study aims to evaluate the systemic cell-mediated immunity and its clinical significance in patients with hepatocellular carcinoma.The influence of liver cancer resection on cell-mediated immunity was also analyzed.Methods:By flow cytometric ananlysis the presence of peripheral blood T lymphocyte subsets in HCC patients and healthy donors were evaluated. The dynamic presence of peripheral T lymphocyte subsets before and after liver cancer resection was also studied.With enzyme-linked immunosorbent assay,the serum cytokines expression of HCC patients and healthy donors were measured.The relationship between the presence of T lymphocyte subsets,cytokine expression and HCC biologic characteristics and TNM stages were analyzed.Results:Compared with the healthy donors,there is significantly abnormal presence of peripheral T lymphocyte subsets,which is related with the HCC biology and TNM stages.CD4+/CD8+ ratio was negatively correlated with the frequency of CD4+CD25+ regulatory T lymphocyte and CD3+T lymphocyte.There was a significant positive correlation between the frequency of CD4+CD25+ regulatory T lymphocyte and CD3+CD45RO+T lymphocyte.Foxp3 positive T lymphocyte may downregulate the expression of CD8+GranzymeB+T cells.It is more specific and reasonable to denote regulatory T cells by CD4+CD25+Foxp3+ T lymphocytes than CD4+CD25+T lymphocytes.There is also a Th1/Th2 serum cytokine imbalance profile in HCC patients and the imbalance is related with HCC biology.There was a significant positive correlation between the expression of peripheral interleukin-10 and interferon-gamma which may imply that interleukin -10 can function as a immunostimulative factor in some conditions and there may be a synergy between the biologic function of peripheral interleukin-10 and interferon-gamma.Our results suggest that liver cancer resection may lead to a temporal enhancement of immunosuppression.However liver cancer resection may reduce the tumour-related regulatory T cells by abating the tumour load.There is a tendency to improve in cell-mediated immunity about one month after sugery.Conclusions:The cell-mediated immunity in HCC patients is in a immunosuppressive condition.The expression of peripheral T lymphocyte subpopulations could be served as the index to evaluate cellular immunity in hepatocellular carcinoma.The abnormal increase of transcription factor Foxp3 expression in peripheral lymphocyte may play an important role in regulate the immunosuppression in hepatocellular carcinoma.The immunosuppressive condition is related with the invasion and progression of HCC.Aberrantly increased peripheral regulatory T lymphocytes may play an important role in the early stage of hepatocellular carcinoma progression.Liver cancer resection may be helpful for the recovery of anti-tumour immunity.The perioperative period of HCC patients may be a beneficial window-phase for immune intervention and regulatory T cells may be served as a target cell.更多还原