权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Investigation of novel therapeutic approach through regulatory T cells (Tr) and dendritic cells (DCs) for the patients with autoimmune hepatitis (AIH)

研究通过调节性 T 细胞 (Tr) 和树突状细胞 (DC) 治疗自身免疫性肝炎 (AIH) 患者的新方法

基本信息

批准号：
18590754
负责人：
OKUMURA Akihiko
金额：
$ 2.5万
依托单位：
Aichi Medical University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

项目摘要

(1) Expression of toll-like receptor (TLR) family on Dendritic cells (DCs) in patients with autoimmune hepatitis (AIH). Total RNA was extracted from isolated T-reg and non-T-reg fractions and the amount of mRNA for TLR2, 3, 4, 6, 7, 8, 9were evaluated by real time PCR. TLR3, 6, 7, 9were expressed on circulating myeloid DC (MDC) and plasmacytoid DC (PDC) in controls, while only TLR6was expressed on those in patients with AIH. Thus different expression pattern of TLRs on DCs might have some influence on the pathogenesis of AIH.(2) Expression of apoptosis-related molecules on DCs, and susceptibility of DCs to apoptosis induced through TNF-R1 in patients with AIH.Total RNA was extracted from circulating myeloid DC (MDC) and plasmacytoid DC (PDC), the amount of mRNA for TNF-R1, TNF-RS18, Fas, Bcl-2, Bax, Bad, FAST, PKR, Apaf-1, and c-FLIP were evaluated by real time PCR. Further, after incubation of MDC+PDC in the presence of TNFα, susceptibility to apoptosis induced through TNF-R1 was evaluated by counting the number of apoptotic DCs. TNF-RS18, TNF-R1, Bad, and FAST were expressed on freshly isolated MDC and PDC both in AIH and control. However, expression of TNF-R1 and Bad was significantly suppressed in AIH compared with controls. After 72 hour incubation in the presence of TNFα, the amount of apoptotic MDC and PDC in controls increased from 30.0% to 45.8%, whereas that in AIH changed from 33.3% to 34.5%, suggesting the possibility that apoptosis of MDC and PDC induced by signals through TNF-R1 is suppressed in AIH.

(1)自身免疫性肝炎（AIH）患者树突状细胞（DCs）上TLR家族的表达从分离的T-reg和non-T-reg组分中提取总RNA，并通过真实的时间PCR来评估TLR 2、3、4、6、7、8、9的mRNA的量。对照组髓系DC（MDC）和浆细胞样DC（PDC）表达TLR 3、6、7、9，AIH组仅表达TLR 6。因此，DCs上TLRs的不同表达模式可能对AIH的发病机制有一定的影响。(2)从循环髓样DC（MDC）和浆细胞样DC（PDC）中提取总RNA，采用真实的时间PCR检测DC中TNF-R1、TNF-RS 18、Fas、Bcl-2、Bax、Bad、FAST、PKR、Apaf-1和c-FLIP的mRNA表达。此外，在存在TNFα的情况下孵育MDC+PDC后，通过计数凋亡DC的数量来评估对通过TNF-R1诱导的凋亡的敏感性。TNF-RS 18、TNF-R1、Bad和FAST在AIH和对照组的新鲜分离的MDC和PDC上表达。然而，TNF-R1和Bad的表达在AIH中与对照组相比显著抑制。TNFα作用72 h后，对照组MDC和PDC凋亡率由30.0%上升到45.8%，而AIH组MDC和PDC凋亡率由33.3%上升到34.5%，提示AIH中TNF R1信号诱导的MDC和PDC凋亡可能受到抑制。