Huang Yi-Hong, Feng Sa-Ran, DU Bing, Xu Kai-Lin, Pan Xiu-Ying
Department of Hematology, The Affiliated Hospital of Xuzhou Medical College, Xuzhou 221002, China.
Zhonghua Xue Ye Xue Za Zhi. 2009 Nov;30(11):735-40.
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.
Responder splenocytes from C57BL/6 donor mice (H-2(b)) were co-cultured with stimulator splenocytes from BALB/c recipient mice (H-2(d)) 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 (IL-4, IL-10 and INF-gamma) production. Flow cytometry (FCM) analysis was used to detect allogeneic chimerism.
(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 < 0.05). 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-gamma 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-BMT, with the lowest in group D (P < 0.01). After BMT, IL-4 and IL-10 levels were slightly decreased in group A, their levels were elevated in groups B and C (P < 0.05) and even more significantly increased in group D (P < 0.01). IL-4 and IL-10 levels peaked between day 10 and 15 post-BMT. (3) The average proportion of H-2(b) positive cells in recipient mice was 95% - 100% on day 60 post-BMT, with complete donor-type implantation.
Combination of mPEG-SPA and anti-OX40L McAb can block T-cell activated antigens and co-stimulatory pathway, regulate the T cells differentiation and induce the immune shift of Th0 cells toward Th2 cells. The immune tolerance induced by this method can significantly relieve aGVHD after allo-BMT.
探讨用甲氧基聚乙二醇-琥珀酰亚胺丙酸酯（mPEG-SPA）和抗OX40L单克隆抗体（McAb）对小鼠同种异体骨髓移植（allo-BMT）后急性移植物抗宿主病（aGVHD）进行预处理的预防效果。
将C57BL/6供体小鼠（H-2(b)）的反应性脾细胞与BALB/c受体小鼠（H-2(d)）的刺激脾细胞在有或无抗OX40L McAb存在的情况下共培养7天，随后进行mPEG-SPA化学修饰。然后将供体骨髓细胞加T细胞混合培养物移植到经致死剂量照射的BALB/c小鼠体内。将BALB/c受体小鼠分为四组：A组（allo-BMT对照组）、B组（mPEG-SPA修饰组）、C组（抗OX40L McAb预处理组）和D组（mPEG-SPA与抗OX40L McAb双处理组）。allo-BMT后观察受体的生存时间和生存率。通过皮肤、肝脏和小肠的临床体征及组织学变化评估GVHD。采用酶联免疫吸附测定（ELISA）检测细胞因子（IL-4、IL-10和INF-γ）的产生。采用流式细胞术（FCM）分析检测同种异体嵌合情况。
（1）A组小鼠出现典型的aGVHD临床体征，所有小鼠在BMT后17天内死亡，平均生存时间（AST）为（12.1±5.5）天。B、C和D组小鼠的aGVHD体征较轻，其AST（分别为36.2±24.9、32.0±24.8和44.3±23.2天）均长于A组（P<0.05）。D组AST最长（P<0.05）。BMT后60天B、C和D组的生存率分别为50%、41.7%和66.7%。（2）A组BMT后血清IFN-γ水平升高，在BMT后10至15天达到峰值，而B、C和D组该水平降低，在BMT后10天达到最低点，D组最低（P<0.01）。BMT后，A组IL-4和IL-10水平略有降低，B和C组水平升高（P<0.05），D组升高更显著（P<0.01）。IL-4和IL-10水平在BMT后10至15天达到峰值。（3）BMT后60天受体小鼠中H-2(b)阳性细胞的平均比例为95% - 100%，实现完全供体型植入。
mPEG-SPA与抗OX40L McAb联合应用可阻断T细胞活化抗原和共刺激途径，调节T细胞分化，诱导Th0细胞向Th2细胞免疫偏移。该方法诱导的免疫耐受可显著减轻allo-BMT后的aGVHD。
