延迟并连续小鼠异基因骨髓移植减轻移植物抗宿主病的探讨  

Delayed sequential mouse allogeneic-bone marrow transplantation attenuating graft-versus-host disease

作　　者：王春玲[1] 徐开林[1] 潘秀英[1] 杜冰[1] 

机构地区：[1]徐州医学院附属医院血液科,江苏徐州221002

出　　处：《中华器官移植杂志》2008年第8期475-478,共4页Chinese Journal of Organ Transplantation

基　　金：国家自然科学基金（30170389）

摘　　要：目的探讨延迟并连续小鼠异基因骨髓移植减轻急性移植物抗宿主病（aGVHD）的作用及其机制。方法选择BALB／c（H-2^d）小鼠为受者，C57BL／6（H-2^b）小鼠为供者。受者接受^60Coγ射线全身照射（TBI）后，建立骨髓移植模型。实验分为5组。对照组：TBI后4h经受者尾静脉输注RPMI1640液0．4ml；经典移植组：TBI后4h经受者尾静脉输注供者的骨髓细胞（BMC）和脾细胞（SC）各1×10^7个／0．2ml；连续移植组：TBI后4h、1d、2d和3d分4次经受者尾静脉输注供者的BMC和SC，每次输注量各为2．5×10^6个／0．05ml；延迟移植组：TBI后第4天经受者尾静脉输注供者的BMC和SC各1×10^7个／0．2ml；延迟并连续移植组：TBI后第4、5、6和7天分4次经受者尾静脉输注供者的BMC和SC，每次输注量各为2．5×10^6个／0．05ml。移植后观察受者的临床表现、病理学改变及存活时间，采用酶联免疫吸附试验（ELISA）测定受者外周血中白细胞介素（IL）-2、IL-4、IL-10及γ干扰素（IFN-γ）的水平，应用流式细胞仪测定受者H-2^b细胞、T淋巴细胞亚群及自然杀伤（NK）细胞的百分率。结果对照组和经典移植组受者均在TBI后3周内因造血功能衰竭和／或aGVHD死亡；连续移植组和延迟移植组受者TBI后60d存活率分别为30％和50％；延迟并连续移植组受者TBI后60d的存活率为70％，高于其它4组（P〈0．05）。延迟并连续移植组IL-4、IL-10的表达高于经典移植组（P〈0．05）；IL-2和IFN-γ的表达低于经典移植组（P〈0．05）。延迟并连续移植组TBI后20d时外周血白细胞计数恢复正常，30d时外周血T淋巴细胞亚群和NK细胞的表达恢复正常，60d时H-2^b细胞的百分率为（98．13±1．11）％。结论延迟并连续小鼠异基因骨髓移植能减轻移植后的aGVHD。其作用机制主要是避开了TBI后炎症因子表达的高峰期，下调了炎症因子和Th1类细胞因子的表达，Objective To explore the effect and mechanism of delayed sequential mouse allogeneic-bone marrow transplantation （BMT） attenuating acute graft-versus-host disease （aGVHD）. Methods BALB/c （H-2^d） mice were used as recipients, and C57BL/6 （H-2^b） mice as donors. The model of BMT was established after recipients accepting total body irradiation （TBI） of ^60Co γ ray. Five experimental groups were set up： control group, 4 h after TBI, recipients were infused via tail vein with 0. 4 ml RPMI 1640 liquid; glassic group, 4 h after TBI, recipients were infused with bone marrow cells （BMC） and spleen cells （SC） 1 × 10^7 per 0. 2 ml liquid respectively; sequential group, at 4 h, 1 day, 2 day and 3 day after TBI, BMC and SC （2. 5 × 10^6 per 0. 05 ml liquid each） were infused via tail vein respectively; Delayed group： at day 4 after TBI, BMC and SC （2. 5 × 10^6 per 0. 05 ml liquid each） were infused via tail vein respectively; Delayed sequential group, at day 4, 5, 6 and 7 after TBI, BMC and SC （2. 5 × 10^6 per 0. 05 ml liquid each） were infused via tail vein respectively. The clinical manifestation, pathologic changes and survival rate of recipients were observed after transplantation. The serum levels of cytokines of the recipients including IL-2, IL-4, IL-10 and IFN-γ were detected by ELISA, and the percentage of H-2h cells, T lymphocyte subpopulation and NK cells was measured by flow cytometry. Results Recipients in control and classic groups all died of hemopoiesis failure and/or aGVHD within 3 weeks after transplantation. The 60-day survival rate in sequential and delayed groups was 30% and 50% respectively, but in the delayed sequential group, the 60-day survival rate was 70 %, which was significantly higher than in other groups （P〈0. 05）. The expression levels of IL-4 and IL-10 in the delayed sequential group were significantly higher than in the classic group, while the expression levels of IL-2 and IFN-γ were on the contrary （P〈0. 05）. In the delayed se

关 键 词：骨髓移植 移植物抗宿主病 小鼠 

分 类 号：R457.7[医药卫生—治疗学] R471[医药卫生—临床医学]