同种异基因骨髓移植GVHD动物模型的建立及其病理生理学机制初探  

Establishment and Pathophysiology Study of Allogeneic Bone Marrow Transplantation Model for Graft-Versus-Host Disease

作　　者：刘杰[1] 王永安[2] 钱远宇[1] 孟庆义[1] 

机构地区：[1]解放军总医院急诊科,北京100853 [2]军事医学科学院毒物药物研究所,北京100850

出　　处：《生物技术通讯》2009年第4期513-516,共4页Letters in Biotechnology

摘　　要：目的:建立稳定的异基因骨髓移植GVHD(移植物抗宿主病)动物模型,并初步了解其病理生理学机制。方法:以BALB/c(H-2d)雌性小鼠为受者,接受8Gy致死剂量的60Co全身照射后,输注雄性C57BL/6(H-2b)供鼠的脾细胞和骨髓细胞,观察受鼠的体征、造血功能恢复及生存时间的变化,并进行病理学、嵌合体和T细胞亚群及相关细胞因子的检查。结果:模型组小鼠在移植后出现了典型的GVHD症状;肠、肝、脾、皮肤的病理学分析均属于Ⅳ度GVHD;嵌合体植入成功;以7、14和21d为检测时间点,发现模型组鼠体内T细胞亚群移植后较移植前CD3+CD4+T细胞数量减少,CD3+CD8+T细胞显著升高,CD4+和CD8+T细胞比例严重倒置,随着时间变化比值会逐渐升高,但仍然处于较显著的倒置水平;血清中IFN-γ、TNF-α在移植后+7d表达显著增高,尤其是IFN-γ的表达在+7d达峰值;IL-4和IL-10的水平在移植前后几乎没有变化。结论:建立了稳定的GVHD动物模型;此模型发病过程中,CD8+T细胞介导的CTL细胞毒性作用可能要大于CD4+Th介导的细胞因子效应;IFN-γ、TNF-α炎症因子在GVHD的早期发挥重要作用;IL-4、IL-10的低水平分泌与急性GVHD的高发病率有关。Objective： To establish a stable model of graft-versus-host disease （GVHD） after allogeneic bone marrow transplantation, and study its pathophysiology preliminary. Methods： The donor（male C57BL/6） bone marrow cells plus splenocytes were transfused into the recipient （female BALB/c） mice which were conditioned with 8 Gy ^60Co lethal total body irradiation. Then, general signs, hematopoiesis recovery, survival time, and chimera should be observed; and pathology section, T cell subsets, cytokines should also be detected. Results： There are typical signs of GVHD in our model mice, as weight loss, depilation, diarrhea and hunched posture, et al; WBC〉1×10^9/L when they died; median survival time is the day 22; histopathology of the gut, liver, spleen and skin illuminate that the fourth grad GVHD has occurred; chimera were observed in the recipients; CD3^＋CD4^＋ T cells obviously decreased, while CD3^＋CD8^＋ T cells markedly increased, and the ratio between the CD4^＋ and CD8^＋ T cells were severely inverted contrasting to the pretransplant. The levels of serum IFN-γ,TNF-α markedly increased on day 7 post-transplantation. No significantly changes on levels of IL-4 and IL-10 pre-and post-transplantation. Conelution： A stable GVHD animal model has been established. CD8^＋ T cell-mediated toxicity of the CTL have a greater effect on GVHD pathogenesis than CD4^＋ Th-mediated immune response. IFN-γ, TNF-α inflammatory cytokines play a important role in early GVHD. The low level secretion of IL-4, IL-10 related to the high incidence of GVHD.

关 键 词：异基因骨髓移植 移植物抗宿主病 动物模型 T细胞亚群 细胞因子 

分 类 号：R392.1[医药卫生—免疫学]