腹腔输注耐受性树突状细胞诱导小鼠免疫耐受研究  

Established the model by transfused tolerogenic dendritic cells in abdominal cavity to induce immunological tolerance in the mouse

作　　者：钟志宏[1] 鄢俊[2] 黄先豹[3] 陈国安[3] 袁利亚[4] 刘振优[1] 

机构地区：[1]赣南医学院,江西赣州341000 [2]赣南医学院第一附属医院,江西赣州341000 [3]南昌大学第一附属医院,江西南昌330006 [4]江西省医学科学研究所,江西南昌330006

出　　处：《中国现代医学杂志》2009年第15期2289-2293,共5页China Journal of Modern Medicine

基　　金：国家自然科学基金(30260039);江西省自然科学基金(2007GZY2599)资助项目

摘　　要：目的通过腹腔输注体外培养的耐受性树突状细胞(tolerogenic dendritic cells,tDCs),在小鼠体内建立tDCs诱导特异性免疫耐受方法。方法采用雄性Balb/c小鼠为供体,雌性昆明小鼠为受体。取供体小鼠骨髓细胞,分离单个核细胞。在含GM-CSF、IL-4、VIP、DXM和LPS等因子的RPMI1640培养基中按不同的组合分为两组进行体外诱导培养:常规DC组(GM-CSF+IL-4)和tDC组(GM-CSF+VIP+DXM)。实验分4组:空白对照组(不经任何处理)、骨髓细胞组(输骨髓细胞)、常规DC组(输常规培养DC)和tDC组(输tDC)。腹腔输注细胞3d后移植供体脾片,移植后20d取移植脾片病理组织学检查,移植后1个月流式细胞术检测受体脾单个核细胞CD80、CD86、CD40、CD11c和CD4+CD25+的表达。结果小鼠体内tDCs输注实验:tDC组的移植脾片大体形态呈灰红色,与移植部位周围组织建立血液供应;病理学观察组织结构完整,未见明显变性坏死,与正常脾组织结构相似。而空白对照组、骨髓细胞组、常规DC组移植脾片大体呈灰白色、浅灰红色和灰褐色,与周围组织未建立血液供应;病理学观察组织结构受破坏,有的呈大片坏死。tDC组小鼠脾单个核细胞CD4+CD25+双表达细胞为(18.15±0.66)%,比正常小鼠的(6.5±0.55)%高近3倍,差异有显著性(P<0.05);CD40、CD11c、CD80和CD86的表达比正常小鼠低,差异有显著性(P<0.05)。结论在移植脾片小鼠模型,经腹腔注射tDCs能诱导受体对移植器官产生特异性免疫耐受。小鼠体内输注tDCs,其脾脏CD4+CD25+Treg细胞增高,CD11c、CD40、CD80和CD86表达降低。腹腔注射tDCs安全性较好,并能形成稳定的嵌合,且维持1个月以上。[Objective] Transfuse tolerogenic dendritic cells （tDCs） cultured in vitro in abdominal cavity, in the mouse established the method of tDCs to induce specific immunological tolerance in vivo. [Methods] Male Balb/c mice are used as donor and female Kunming mice as receptor. Tolerogenic dendritic cells were generated from bone marrow precursors cultured with GM-CSF, IL-4, VIP, DXM and LPS. The cells were divied into two groups： common-DC group （GM-CSF＋IL-4）, tDC group （GM-CSF＋VIP＋DXM）. The animal transfusion experiment were divided into four groups： Blank group （not with any treatment）, bone marrow group （transfusion bone marrow）, common-DC group （transfusion common-DC）, tolerogenic-DC group （transfusion tolerogenic-DC）. Three days after transfusion transplant the pieces of donor＇s spleen to receptor. Twenty days after transplantation take the pieces of transplanted spleen into pathological section. A month after transplantation detect CD80, CD86, CD40, CD11c and CD4^＋CD25^＋ of spleen single caryoceils of receptor through FCM. [Results] The ceils possessed morphological character of typical DC. In the transfusion experiment of mouse, the tDC group present grayish red, and establish blood supply to surrounding structure; in the histopathologic examination it didn＇t see obviously denaturation and necrosis. But the Blank group, bone marrow group, common-DC group present grayish white, light grayish red, grayish brown, and didn＇t establish blood supply to surrounding structure; in the histopathologic examination it see obviously necrosis. The CD4^＋CD25^＋ cell of tDC group is （18.15±0.66）%,and the normal mouse is （6.5±0.55）% （P 〈0.05）. Compared to the common-DC group, the tDC group lowly express CDsc, CDs6, CD4o and CDnc （P 〈0.05）. [Conclusions] Transfused tDC to the mouse induce specific immunological tolerance in vivo. The CD,~CD~~ cell of tDC group is high,and lowly express CD80, CD86, CD40 and CD11c. The safety of transfused tDCs in abdominal

关 键 词：树突状细胞 免疫耐受 小鼠 调节性T细胞 

分 类 号：R392.4[医药卫生—免疫学]