【作者】 孙文学；

【导师】 戴勇；

【作者基本信息】 暨南大学 ， 内科学， 2004， 硕士

【摘要】 目的 探讨慢性肾功能衰竭(CRF)患者外周血树突状细胞(DC)数量及活性，为监测细胞免疫功能及评估腹膜透析、血液透析对机体免疫状态的影响提供直接敏感的新指标，为开展CRF免疫治疗奠定基础。 方法 CRF患者41例和正常人23例，患者按病情分期及治疗与否分为：氮质血症组、尿毒症组(未透析)、腹膜透析组(PDG)、血液透析组(HDG)。进行以下方面的研究： (1)人DC分类试剂盒检测外周血DC亚群百分率。 (2)分离外周血单个核细胞，细胞因子(GM-CSF、IL-4、TNFet)体外诱导培养，流式细胞仪检测DC表型分子表达水平。 (3)ELISA法检测第9天DC培养上清液中IL-12的含量。 (4)异体混合淋巴细胞反应(AMLR)评估DC刺激T细胞增殖的能力。 结果 (1)DC亚群状况：与正常组比较，氮质血症组总DC、髓系树突状细胞(MDCl)增加(P＜0.05)；尿毒症组总DC、淋巴系树突状细胞(PDC)减少(P＜0.01)。HD组MDCl减少，PDC增加，MDCl／PDC降低(P＜0.01)。 (2)DC表型分析：与正常组比较，尿毒症组DC表型分子表达均减少(P＜0.01)：PD组DC表达CD40、CD80、CD83减少(P＜0.05)；HD组DC表达CD1α、CD11c、CD40较低,CD123较高(P＜0.05)，CD80、CD83接近正常。 (3)IL－12含量检测：氮质血症组DC分泌IL－12明显增加，与其它各组差异显著，P均为0.000；尿毒症组、PD组和HD组DC分泌IL-12明显减少(P＜0.05)。 (4)AMLR：与正常组比较，DC刺激T细胞增殖的能力在尿毒症组和PD组减弱，氮质血症组增强(P＜0.01)，HD组接近正常(P＞0.05)。 结论 CRF患者存在外周血DC数量及功能紊乱，氮质血症期代偿性数量增多且活化，暨南大学硕士学位论文尿毒症期数量减少且功能缺陷:维持性腹膜透析患者DC数量改善，但功能仍低下;而维持性血液透析患者MDCI减少，PDC增加，MDCI/P DC倒置，DC功能接近正常。肾脏替代治疗只能部分改善DC活性。更多还原

【Abstract】 ObjectiveTo demonstrate the cellular immune function, evaluate the influnce of peritoneal dialysis and hemodialysis on the immune status of patients with chronic renal failure (CRF) and establish the foundation for the development of immunotherapy on CRF, the quantity and activity of dendritic cell (DC) of peripheral blood in patients with CRF were counted and analysed in this experiment.MethodsForty-one patients with CRF and 23 healthy individuals were used in this study, According to the patients’ condition and intervention, patients were divided into four groups: azotemia group and uremic group (CRF patients on conservative treatment), peritoneal dialysis group (PDG) (uremic patients on regular peritoneal dialysis), hemodialysis group (HDG) (uremic patients on regular hemodialysis).(1) The frequency of subsets of DC was detected by human Blood DC enumeration Kit.(2) The peripheral blood mononuclear cells (PBMCs) of T cell-depleted populations were incubated and induced into mature DCs in RPMI-1640 medium in the presence of cytokines GM-CSF, IL-4, TNF-a and 100ml.L(-1 ) of FBS for a total of 9 days in vitro experiment. The expressions of surface marker on DCs were detected by flow cytometry.(3) ELISA was used to detect the cytokine level of IL-12 in the supernatant produced by DCs on the 9th day.(4) The stimulatory capacity of DCs to T cell proliferation was evaluated by allogenic mixed lymphocyte reaction (AMLR).Results(1) Analysis of the quantity of subsets of DC: The quantities of DC and meyloiddendritic cell (MDC)l from azotemia group increased, while the quantities of DC and plasmacytoid dendritic cell (PDC) from uremic group decreased (P<0.05) ; the quantity of MDC1 from HDG significantly decreased, while PDC increased leading to the proportion of them inverse compared with healthy controls(P<0.01).(2) Analysis of phenotype of DC: Compared with healthy controls, the expression levels of phenotype markers on DC surface from uremic group significantly decreased (P<0.01); the expression levels of CD40, CD80,CD83 on DC surface from PDG decreased (PO.05); the expression levels of CDla,CDllc,CD40 on DC surface form HDG decreased(P<0.05),while that of CD 123 increased except for the normal expression levels of CD80,CD83.(3) Detection of IL-12: The production of IL-12 of DCs from uremic patients was decreased, so did in PDG and HDG patients, while that of DCs from azotemia patients was increased compared with healthy controls (P < 0.05).(4) AMLR: The stimulatory capacity of DCs to T cell proliferation from uremic patients and PDG patients significantly decreased, while that of DCs from azotemia patients significantly increased in AMLR (P<0.01). The stimulatory capacity of DCs to T cell proliferation from HDG patients was approach to normal compared with healthy control (P>0.05).ConclusionThe patients with CRF had the immature phenotype of DC that was defect in fuction and decreased in quantity. The quantity and activity of DC increased in azotemia stage but decreased in uremic stage; DC from PD patients had defective function, while ameliorated in quantity; The quantity of MDC from HD patients remain to be reduced but PDC increased leading to the proportion of them inverse, while they had normal function. Our results indicated that kidney replacement therapy could partly ameliorate the metabolism of DC.更多还原