移植物化学修饰与阻断OX40一OX40L途径预防异基因骨髓移植小鼠移植物抗宿主病的研究  

Prophylaxis of graft-versus-host disease in mice by chemical modification of graft and OX40-OX40L costimulatory pathway

作　　者：黄一虹[1] 冯洒然[1] 杜冰[1] 徐开林[1] 潘秀英[1] 

机构地区：[1]徐州医学院附属医院血液科,221002

出　　处：《中华血液学杂志》2009年第11期735-740,共6页Chinese Journal of Hematology

基　　金：基金项目：江苏省高校自然科学基金（07KJD320224）

摘　　要：目的观察甲氧基聚乙二醇-琥珀酰亚胺丙酸酸酯（mPEG-SPA）与抗OX40L单抗预处理移植物对小鼠异基因骨髓移植后急性移植物抗宿主病（aGVHD）的影响及其机制。方法供、受鼠的脾淋巴细胞体外混合培养，加入抗OX40L单抗孵育培养后，再用mPEG—SPA化学修饰，与C57BIM6供鼠骨髓细胞混合后移植给经致死量全身照射的BALB／c受鼠。同时设相应的对照组。观察移植后受鼠aGVHD的临床表现、病理学改变、存活时间及生存率等，检测移植鼠T淋巴细胞亚群，细胞因子的变化及异基因嵌合体。结果①单纯骨髓移植对照组小鼠均出现aGVHD临床表现，17d内全部死于aGVHD，存活时间为（12．1±5．5）d；而mPEG—SPA和抗OX40L单抗单独或联合预处理组小鼠一般状态较好，部分出现aGVHD表现，且较单纯骨髓移植对照组症状轻，皮肤、肝脏、小肠病理表现均较单纯骨髓移植对照组明显减轻，mPEG—SPA和抗OX40L单抗单独或联合预处理组小鼠存活时间分别为（36．2±24．9）、（32．0±24．8）和（44．3±23．2）d，均较单纯骨髓移植对照组明显延长（P〈0．05），其中抗OX40LmPEG—SPA联合预处理组小鼠平均存活时间最长（P〈0．05），mPEG—SPA单独预处理组和抗OX40L单独预处理组生存率分别为50．0％、41．7％，明显高于单纯骨髓移植组，OX40LmPEG—SPA联合预处理组生存率最高，为66．7％。②移植后对照组小鼠血清IFN一吖浓度升高，在＋10～＋15d时达高峰；而mPEG—SPA和抗OX40L单抗单独或联合预处理组降低，＋10d时降至最低，OX40LmPEG—SPA联合预处理组降低最明显（P〈0．01）。移植后对照组IL4、IL一10浓度轻度降低，而mPEG-SPA和抗OX40L单抗单独或联合预处理组均明显升高（P〈0．05），＋10～＋15d达高峰，以联合组升高最明显（P〈0．01）。@mPEG—SPA和抗OX40L单抗单独或联合预处理组＋60d时异基因嵌合率�Objective To explore the prophylaxis effect of pretreatment of allograft with methoxypolyethylene glycol-succinimidyl-propionic acid ester （mPEG-SPA） and anti-OX40L monoclonal antibody （McAb） on acute graft-versus-host disease （aGVHD） after allogeneic bone marrow transplantation （allo- BMT） in mice. Methods Responder splenocytes from C57BL/6 donor mice （H-2h） were co-cultured with stimulator splenocytes from BALB/c recipient mice （ H-2d ） for 7 days in the presence or absence of anti- OX40L McAb followed by mPEG-SPA chemical modification. Donor bone marrow cells plus the mixed culture of T-cells were then transplanted into lethally irradiated BALB/c mice. The BALB/c recipient mice were divided into four groups： group A （allo-BMT control group）, group B （mPEG-SPA modification group）, group C （anti-OX40L McAb pretreated group） and group D （mPEG-SPA and anti-OX40L McAb dual-treated group）. Survival time and survival rate of the recipients were observed after allo-BMT. GVHD was assessed by clinical signs and histological changes of skin, liver and small intestines. Enzyme-linked immunosorbent assay （ELISA） was used to detect cytokines （IL4, IL-10 and INF-γ） production. Flow cytometry （FCM） analysis was used to detect allogeneic chimerism. Results （1）The mice in group A developed typical clinical signs of aGVHD and all mice died within 17 days after BMT with an average survival time（AST） of （ 12.1 ± 5.5） days. The signs of aGVHD were less evident in mice of groups B, C and D, and their AST （36.2 ± 24.9, 32.0 ± 24.8 and 44.3 ± 23.2 days, respectively） were all longer than that in group A （ P 〈 O. 05 ）. AST of group D being the longest （P 〈 0.05 ）. The survival rates at day 60 post-BMT in groups B, C and D were 50%, 41.7% and 66.7%, respectively. （2）Serum IFN-γ level was increased after BMT in group A, and peaked in day 10 to day 15 post-BMT, while the level was decreased in groups B, C and D, reached the nadir on the day 10 post

关 键 词：甲氧基聚乙二醇 化学修饰 抗原 抗体 OX40L 移植物抗宿主病 

分 类 号：R686[医药卫生—骨科学]