Toll样受体信号通路在小鼠皮肤移植免疫反应中的作用  

Role of toll-like receptor signaling in skin transplantation responses in mice

作　　者：李超[1] 黄霞[1] 杨涛[1] 张利民[1] 李明强[1] 周平[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院器官移植研究所,武汉430030

出　　处：《中华器官移植杂志》2013年第9期550-553,共4页Chinese Journal of Organ Transplantation

基　　金：国家自然科学基金（81072442）

摘　　要：目的研究树突状细胞（DC）中Toll样受体（TLR）信号通路在小鼠皮肤移植排斥反应中的作用。方法体外诱导培养得到BALB／c小鼠骨髓源性的不成熟DC，加入TLR激动剂CpG刺激细胞成熟，实验组同时加入TLR通路关键分子MyD88的抑制剂ST2825，24h后用流式细胞仪检测IX2共刺激分子CD80和CD86的表达；用荧光染料CFSE标记的C57BL／6小鼠T淋巴细胞与BALB／c小鼠的DC在CpG刺激下行混合淋巴细胞培养，实验组同时加入ST2825，培养3d后用流式细胞仪检测T淋巴细胞的增殖情况。建立BALB／c小鼠次要组织相容性抗原HY错配皮肤移植模型，供鼠均为雄性小鼠，受鼠均为雌性小鼠，在此基础之上分为对照组（供受鼠均为野生型BALB／c小鼠）、MyD88组（供受鼠均为MyDS8基因敲除的BALB／c小鼠）、DC组（供受鼠均为MyD88基因敲除的BALB／c小鼠，移植后输注供鼠来源的DC）、实验组（供受鼠均为MyD88基因敲除的BALB／c小鼠，输注经ST2825预处理的供者来源DC）。观察各组移植皮片的存活时间。结果TLR通路抑制剂ST2825可以剂量依赖性地抑制CpG刺激引起的DC共刺激分子CDS0和CD86的高表达。经Cpg刺激的IX；可以诱导同种反应性T淋巴细胞的大量增殖，ST2825可以呈剂量依赖性地抑制此反应。对照组移植物的存活时间为（22．8±2．8）d；MyD88组移植物均未发生排斥反应（存活时间超过100d）；DC组移植物存活时问为（9．7±2）d；实验组移植物亦未发生排斥反应（存活时间超过100d），较DC组显著延长（P〈0．05）。结论DC通过TLR信号通路在移植排斥反应中发挥关键作用，TLR信号通路抑制剂ST2825能够抑制DC的激活和生物学功能，其机理可能与ST2825抑制DC成熟，并间接抑制同种反应性T淋巴细胞的增殖有关。Objective To investigate the essential role of Toll-like receptor （TLR） signaling in dendritic cells in rejection responses of skin transplantation in mice. Method Immature wild type BALB/c bone marrow dentritic cells （BIVIDCs） were stimulated by a TLR agonist, CpG, in the presence of ST2825, an inhibitor of key molecule myeloid differentiation factor 88 （MyD88） in TLR signal transduction, or in the absence for 24 h in vitro. Co-stimulatory molecules CDS0 and CD86 were analyzed by flow cytometry. Wild-type C57BL/6 na ve T cells stained with CFSE, a fluorescent dye, were co-cultured with BALB/c DCs for 3 days in the presence or absence of ST2825. CD3±/CFSE± cells were analyzed by flow cytometry. Minor antige-mismatched （HY-mismatched） skin allograft model was employed in this study in which the donors were all males and the recipients were all females. The experimental groups were divided as follows-, the control group, the donors and the recipients were all wide type BALB/c mice; the MyD88 group, the donors and the recipients were all MyD88-konckout BALB/c mice; the DCs group, the donors and the recipients were all MyD88- knockout BALB/c mice and the recipients were transfused with donor-sourced DCs after transplantation; the experimental group, the donors and the recipients were all MyD88 knockout BALB/c mice and the recipients were transfused with dono〉sourced DCs pre-treated with ST2825 after transplantation. The survival time of skin allografts was observed. Result ST2825 could dose-dependently inhibit the high level expression of co-stimulatory molecules CD80 and CD86 induced with CpG. Similarly, and it could also dose-dependently inhibit the allo-reactive T cell proliferation upon the stimulation with DCs. The survival time of every group was as follows： the control group （22. 8 ± 2.8） days vs. the MyD88 group （〉100 days） vs. the DCs group （9. 7 ± 2） days vs. the experimental group （〉100 days, P〈0.05, vs. the DC group）. Conclusion DCs may play an essential role

关 键 词：小鼠 皮肤移植 树突状细胞 TOLL样受体 

分 类 号：R392[医药卫生—免疫学]