降低成人胰岛免疫原性的研究  

Investigation on lowering immunogenesis of islet grafts from human adult

作　　者：董维平[1] 陈向峰[2] 彭永德[3] 李永翔[2] 王煜非[1] 王鉴波[1] 胡远峰[1] 

机构地区：[1]上海交通大学附属第一人民医院糖尿病研究室,200080 [2]上海交通大学附属第一人民医院肾移植泌尿科,200080 [3]上海交通大学附属第一人民医院内分泌科,200080

出　　处：《中华器官移植杂志》2006年第3期145-148,共4页Chinese Journal of Organ Transplantation

基　　金："上海市临床医学器官移植中心"基金资助(ZX01A06)

摘　　要：目的探讨体外预处理对成人胰岛移植物免疫原性的影响。方法实验分三组进行,分离并纯化成人胰岛,对照组胰岛在37℃下培养2 d;24℃组胰岛在24℃下培养2 d;抗体组胰岛在37℃下培养1 d后,加抗人主要组织相容性复合物Ⅱ类单克隆抗体(抗MHCⅡ类单抗)和补体再培养1 d。采用胰岛-淋巴细胞混合培养(M ILC)、抗HLA-DR单克隆抗体和抗人白细胞共同抗原(LCA)单克隆抗体比较胰岛的免疫原性,经氚(3H)-亮氨酸掺入、胰岛素释放试验和胰岛细胞凋亡检测观察胰岛的功能和活力。结果抗体组M ILC刺激指数较对照组明显降低(P<0.05),而24℃组虽低于对照组,但差异无统计学意义(P>0.05)。抗体组和24℃组HLA-DR阳性细胞、LCA阳性细胞较对照组明显降低(P<0.01)。抗体组和24℃组低糖刺激下胰岛素分泌量、高糖刺激下胰岛素分泌量、胰岛素释放指数3、H-亮氨酸掺入率(闪烁计数值)及细胞凋亡率与对照组相比,差异均无统计学意义。结论成人胰岛经抗MHCⅡ类单抗预处理或短期24℃培养,能明显降低其免疫原性,同时对胰岛的功能无明显影响。Objective To lower irnmunogenesis of human adult islets, reduce the dose of immunosuppressors and promote wide development of transplantation of adult islets, isolated adult islets were in vitro pretreated. Methods Isolated adult islets were cultured at 24 ℃ for 2 days （24 ℃ group） or pretreated using MHC-Ⅱ monoclonal antibody （monoclonal antibody and complements were added and cultivated for another day following one-day-24 ℃-culture, antibody group） as observation groups. Adult islets were cultured at 37 ℃ in CMRL 1066 medium plus 20 % fetal bovine serum for 2 days as control. The immunogenics of the islets was detected using lymphocyte-islet mixed culture （MILC） and immunohistochemical staining of HLA-DR and lymphocyte common antigen （LCA）. The function and activity of the islets were identified using 3 H-leucine incorporation assay, insulin release test and in situ apoptosis assay. Results Compared with control, MILC stimulation index was remarkably lower in the antibody group （P〈0. 05）, and low in the 24 ℃ group, but without statistic difference （P〉0. 05）. The percentage of HLA-DR and LCA positive cells in the antibody groupand in the 24 ℃ group was much lower than that in control （All P〈0. 01 ）. However, there were no significant difference in 3 H-leucine incorporation, insulin release level and number of apoptotic cells among three groups. Conclusions HLA-DR positive cells and lymphocytes are decreased, immunogenics of islet grafts reduced, and the function of the islets keeps in good condition after pretreatment of MHC-Ⅱ monoclonal antibody and short-time culture at 24℃.

关 键 词：胰岛移植 移植预处理 免疫 

分 类 号：R657.5[医药卫生—外科学]