供者肝内单个核细胞与肝移植后急性排斥反应的相关性研究

摘要

目的 研究供者肝移植物内的单个核细胞(intrahepatic mononuclear cell,HMC)的表型分析和产生的细胞因子,及其对移植物急性细胞排斥反应(acute cellular rejiection,ACR)的影响.方法 采集2006年4月-2010年8月伦敦国王学院医院行原位肝移植的60例供者的HMC,采用流式细胞分析技术检测其表面免疫分子标志物和细胞因子,将其与上述移植物受者ACR的发生率进行相关性分析.结果 HMC内CD4+/CD8+比值(均值0.5)明显低于健康者外周血比例(均值1.4),证实细胞标本的肝源性.23.33%的患者在随访期发生了ACR.发生反应患者HMC内的自然杀伤(nature killer,NK)细胞频率[(24.65±4.83)%]明显低于无排斥反应患者[(39.46±2.93)%],P=0.020,呈负相关(r=-0.321).发生ACR患者HMC内产白细胞介素(interleukin,IL)-2的CD4+T淋巴细胞水平[(20.55±2.29)%]明显高于无排斥反应患者[(14.95±1.29)%],P=0.048;产IL-4的CD4+T淋巴细胞水平[(6.5±0.84)%]高于无排斥反应患者[(3.85±0.41)%],P=0.006;产IL-17的CD8+T淋巴细胞水平[(5.39±1.17)%]高于无排斥反应患者[(2.25±0.31)%],P=0.025;产IL-4的CD8+T淋巴细胞[(7.05±0.73)%]高于无排斥反应患者[(4.51±0.56)%],P=0.041;产IL-23的CD8+T淋巴细胞[(3.0±0.64)%]高于无排斥反应患者[(1.74±0.24)%],P=0.031.结论 供肝内的NK细胞、产IL-2、IL-4的CD4+T淋巴细胞、产IL-17、IL-4和IL-23的CD8+T淋巴细胞与肝移植患者发生ACR相关,上述细胞可能参与ACR的发生.%Objective To investigate the phenotypic analysis and cytokines from hepatic mononuclear cells (HMCs) in liver grafts and their influence on acute cellular rejection (ACR) after liver transplant. Methods HMCs from liver grafts of 60 patients undergoing primary liver transplant in King's College Hospital in London from Apr. 2006 to Aug. 2010 were collected. Flow cytometry was used to detect the surface markers and cytokines. Correlation analysis was done between ACR and the surface markers and cytokines. Results CD4＋/CD8＋ T lymphocyte ratio in HMCs (mean: 0.5) was lower than that in peripheral blood monuclear cells in healthy controls (mean: 1.4), continning the hepatic origin of cell samples. ACR occurred in 23.33％ of the patients during the follow-up. The frequency of natural killer (NK) cells in the patients with ACR (24.65％±4.83％) was lower than that in the patients without ACR (39.46％±2.93％) (P＝0.020). The correlation coefficient between the incidence of ACR and NK cell frequency in HMCs was-0.321.The frequencies of CD4＋ T lymphocytes producing interleukin (IL)-2 and IL-4 in HMCs of the patients with ACR were 20.55％e2.29％ and 6.5％±0.84％, higher than those in the patients without ACR (14.95％±1.29％ and 3.85％±0.41％, respectively). The frequencies of CD8＋ T lymphocytes producing IL-17, IL--4 and IL-23 in HMCs of the patients with ACR were 5.39％±1.17％, 7.05％±0.73％ and 3.0％±0.64％, higher than those in the patients without ACR (2.25％±0.31％, 4.51％±0.56％ and 1.74％±0.24％, respectively).Conclusions NK cells, CD4+ T lymphocytes producing IL-2 and IL-4, and CD8+ T lymphocytes producing IL-17, IL-4 and IL-23 in liver grafts are associated with the incidence of ACR after liver transplant. These cells may participate in the incidence of ACR.