肝病患者的肝组织单细胞悬液与外周血中淋巴细胞亚群的分析比较

目的探讨肝脏组织单细胞悬液的制备方法,了解慢性肝病患者的肝组织和外周血的淋巴细胞亚群差异。方法分别选择慢性乙型肝炎(CHB)14例、自身免疫性肝炎(AIH)4例和原发性肝细胞癌(HCC)3例,在超声引导下经皮作肝脏穿刺活检,用单细胞制备仪或物理研磨方法将肝组织制备成单细胞悬液,肝组织中淋巴细胞经4种特异性荧光抗体标记,在流式细胞仪(FCM)上作四色荧光分析,并与外周血测定结果比较。结果单细胞制备仪和物理研磨方法制备的肝组织单细胞悬液,细胞成活率分别为92.4%和91.5%;检测肝组织单细胞悬液中淋巴细胞亚群,正常对照组和慢性肝病各组间差异均有显著性(P<0.05),AIH组和CHB组间差异有显著性(P<0.05)。而外周血中淋巴细胞亚群检测,HCC组和CHB组间差异有显著性(P<0.05)。肝组织和外周血相应项目比较差异均有显著性(P<0.05)。结论用2种方法制备单细胞悬液能满足FCM检测的特殊需要;肝脏组织中淋巴细胞与外周血的淋巴细胞亚群比例差异有显著性。

Study on the difference between the lymphocyte subsets of liver tissue and peripheral blood in liver disease patients

Objective To study the approaches for obtaining the cell suspension from liver biopsy specimens and compare the difference of lymphocyte subsets between peripheral blood and liver tissue in the patient with chronic liver disease. Methods Fourteen chronic hepatitis B (CHB),four autoimmune hepatitis(AIH) and three hepatic cell cancer (HCC) patients were studied.The liver tissue was obtained by ultrasound guided liver biopsy.The cell suspension was prepared by medimachine and mechanical dissociation from liver biopsy specimen.Then the final cell suspension was analysed by 4-color flow cytometry. Results The mean viability of medimachine method and mechanical dissociation method was 92.4% and 91.5% respectively.The liver lymphocyte subsets proportions showed significant difference between each chronic liver disease group and normal control group,and also between the group of AIH and CHB (P<(0.05)).The peripheral lymphocyte subsets proportions showed significant difference between the group of HCC and CHB (P<0.05).The proportions of lymphocyte subsets of peripheral blood and liver tissue showed significant difference (P<0.05). Conclusions The two approaches of preparing the hepatic cell suspension are both suitable to make flow cytometric analysis.There are difference in the proportion of lymphocyte subsets between in peripheral blood and in liver tissue.