【作者】 陶红；

【导师】 李秋；

【作者基本信息】 重庆医科大学 ， 儿科学， 2010， 硕士

【摘要】 第一部分过敏性紫癜发病中树突状细胞与T辅助细胞分化失衡的关系目的:探讨树突状细胞(DC)调控信号的异常与过敏性紫癜(HSP)及过敏性紫癜肾炎(HSPN)病人发病中T辅助细胞分化失衡的关系。方法:分别采用散射比浊法、ELISA和流式细胞技术,检测HSP及HSPN患儿血浆中免疫球蛋白、血浆IL-4、INF-γ和DC培养上清IL-12、IL-10水平及DC表面CD86、CD80、CD83、HLA-DR、CD-209的表达情况。结果:与对照组比较,HSP及HSPN组患儿:血浆免疫球蛋白IgA及IgE水平升高;血浆中Th2类细胞因子IL-4升高,Th1类细胞因子INF-γ降低;DC培养上清中IL-10升高,IL-12降低;DC表达CD86、CD83、HLA-DR水平升高,表达CD80水平降低。CD-209在对照组、HSP及HSPN之间差异无统计学意义。结论:DC的抗原特异性信号、协同刺激信号和分化或极化信号三种调控信号的异常,与HSP及HSPN患儿Th1/Th2失衡、免疫球蛋白合成异常密切相关。第二部分NF-κB—DC—Th细胞信号通路在过敏性紫癜免疫发病中的作用目的:了解NF-ΚB—DC—Th细胞信号通路失衡在HSP及HSPN发病中的作用。方法:分别采用ELISA、流式细胞技术、RT-PCR和细胞免疫化学方法,检测对照组、HSP和HSPN患儿未加入NF-ΚB抑制剂BAY-11-7082和加入抑制剂后, DC培养上清IL-12、IL-10水平;DC表面CD86、CD80、CD83、HLA-DR、CD-209的表达;DC内NF-κB亚型p50、p65 mRNA表达量和DC内NF-κB活化情况。结果:与对照组比较,HSP组和HSPN组: DC培养上清中IL-10升高,IL-12降低;CD86、CD83、HLA-DR升高,CD80降低;细胞内NF-κB p50、p65 mRNA表达量也升高; NF-κB活化表达数量增多,但HSP与HSPN以上检测指标无显著差异。加入NF-κB抑制剂后以上各项异常指标可逆转,且逆转后与对照组比较无显著差异。结论:HSP发病与DC异常调控Th1/Th2细胞失衡密切相关,NF-κB是DC免疫功能的关键性调控基因,抑制NF-κB的活化是阻断DC功能和依赖于T细胞免疫反应的一个有效靶点,将成为早期治疗HSP的一条有效途径。更多还原

【Abstract】 PARTⅠTHE RELATION OF DENDRITIC CELL TO THE IMBALANCE DIFFERENTIATION FO T HELPER CELL IN PATHOGENESIS OF HENOCH-SCHONLEIN PURPURAObjective: Discussion the relation between the abnormal regulation signals of dendritic cells(DC) and the dysimmunity of pathogenesis of Henoch-Schonlein purpura (HSP).Methods: Immunoglobulin Levels were detected by nephelom etry assay in children with HSP . Inflammatory cytokines IL-4, INF-γin blood plasma and IL-12, IL-10 in the supernatants of cultured DC were detected by ELISA. The count of CD86, CD80, CD83, HLA-DR, CD-209 expressed on DC were detected by cell flow cytometry.Results: HSP group showed: The levels of plasma IgA and IgE were higher than the control group, IL-12 levels in the supernatants of cultured DC and INF-γlevels in the plasma were lower than the control group,IL-10 levels in the supernatants of cultured DC and IL-4 levels in the plasma were significantly increased than the control group. CD86, CD83, HLA-DR expressed on DC were significantly higher and CD80 was significantly lower than the control group, P<0.05. CD-209 of Two groups had no statistical significance, P>0.05.Conciusion: The anomalies of antigenspecific signals, costimulatory signals and differentiation or polarization signal of DC might be involved in Th1/Th2 imbalance and synthesis of abnormal immunoglobulins in pathogenesis HSP.PARTⅡNF-κB- DC–Th CELL SIGNALING PATHWAY IN THE ROLEOF IN PATHOGENESIS OF HENOCH-SCHONLEIN PURPURAObjective :To investigate The relation between nuclear transcription factor-κB on regulation of dendritic cell and imbalance of T helper cell differentiation in the pathogenesis of Henoch-Schonlein purpura.Methods :Inflammatory cytokines IL-12,IL-10 in the supernatants of cultured DC were detected by ELISA. The cell count of CD86,CD80,CD83,HLA-DR,CD-209 expression on DC were determined by cell flow cytometry. RT-PCR detection of NF-κB isoforms p50,p65mRNA expression.cells immunohistochemical detection the abnormal NF-κB expression in DC. Detection of the control group, HSP and HSPN patients did not join the NF-ΚB inhibitor BAY-11-7082 and added inhibitors, the above indicators of change.Results: Compared with the control group, DC cultured in HSP and HSPN group showed: IL-12 levels in the supernatants of cultured DC lower than the control group,IL-10 levels in the supernatants of cultured DC was significantly increased than the control group;CD86,CD83, HLA-DR was significantly higher but CD80 was significantly lower than the control group.NF-κB p50,p65 mRNA expression intracellular also increased,and had a large number of NF-κB activation. HSP and HSPN group detected had no statistical relationship between the above, When NF-κB inhibitors to join most of the above indicators are back to normal.Conclusion :The function of Th1 was decreased while the function of Th2 was increased in HSP,which were related to DC abnormal discussion.NF-κB is a key regulatory genes of DC immune function , inhibit activation of NF-κB is a valid target to blocking DC function and depends on the T-cell immune response,It will be an effective way to treatment HSP earlier.更多还原