输注人PBMC建立“人—鼠”异种移植物抗宿主病模型

发布时间：2018-04-01 21:34

本文选题：器官移植　切入点：异种移植　出处：《华中科技大学》2011年硕士论文

【摘要】：在人体内的实验由于道德、伦理、法律等诸方面的制约而受到严格限制,涉及人体细胞的研究一般只能进行体外实验,而体外实验的结果往往与体内情况存在一定的差别;“人-鼠”模型的建立,为在体内研究提供了可能。然而目前对人-鼠模型的报道也较多,但行之有效且较稳定的方法报道甚少。可供借鉴的异种移植物抗宿主病模型(graft-versus-host disease,GVHD)以“人-鼠”模型应用最广泛,但建立的方法也有较多的报道。本研究主要探讨选择合适小鼠品系,优化相关实验条件,建立稳定的“人-鼠”异种移植物抗宿主病(X-GVHD)模型,为研究诱导人T细胞耐受提供动物模型。主要内容与结果如下: 本研究选取了裸鼠、NOD/SCID经亚致死剂量γ射线全身照射后,腹腔输注健康人外周血单核细胞(PBMC)建立异种急性X-GVHD模型。通过检测人T细胞在尾静脉血、组织、器官中的浸润情况等指标(采用流式细胞术和免疫组化技术),比较两种模型鼠中人免疫细胞的浸润率,各组生存时间,从而确定建立X-GVHD模型的合适小鼠品系,并摸索人PBMC的输注途径和合适细胞量,以及观察免疫细胞表型变化与功能的最佳时间窗口。结果显示,NOD/SCID鼠更适合诱导“人鼠”X-GVHD模型的小鼠;腹腔输注途径和尾静脉输注途径对建立“人鼠”X-GVHD模型无明显差异;腹腔输注5×107以上人PBMC可以成功建立“人鼠”X-GVHD模型。在采用优化后的实验条件建立模型中,监测人T细胞表型动态变化与功能的最佳时间窗口为7-11天;模型鼠的平均生存时间为14.16±1.77天。本研究的创新点和意义: 本研究使用不同的小鼠建立模型,经检测后,建立了“人-NOD/SCID异种急性移植物抗宿主病模型”,进一步优化各项条件,建立了稳定的“人-鼠”X-GVHD模型,该模型为诱导人T细胞耐受的研究提供了实验依据。同时该模型可以用于体内药敏实验研究,预测指导临床治疗方案的选择和设计,以及探索放疗、内分泌治疗、免疫治疗、基因治疗及分子靶向治疗等新型的治疗手段。
[Abstract]:Experiments in the human body are severely restricted by moral, ethical, legal and other constraints. Studies involving human cells are generally confined to in vitro experiments. However, the results of in vitro experiments are often different from those in vivo, and the establishment of the "human-mouse" model makes it possible to study in vivo. However, there are many reports on the human-mouse model at present. However, there are few reports of effective and stable methods. Graft-versus-host disease (GVHD), a model of xenograft-versus-host disease (GVHD), is the most widely used in the "human-mouse" model. However, there are many reports on the established methods. In this study, the suitable strain of mouse was selected, the experimental conditions were optimized, and a stable "human-mouse" xenograft versus host disease X-GVHD model was established. To provide an animal model for the study of inducing human T cell tolerance. The main contents and results are as follows:. In this study, a xenogeneic acute X-GVHD model was established by intraperitoneal infusion of nod / SCID into healthy human peripheral blood monocytes after whole-body irradiation with sublethal dose 纬 -ray. Human T cells were detected in caudal vein blood and tissue. The infiltration rate of human immune cells and survival time of each group were compared by flow cytometry and immunohistochemical technique, so as to determine the appropriate strain of X-GVHD model. The optimal time window for observation of phenotypic changes and function of immune cells was found. The results showed that Nod / SCID mice were more suitable for inducing "human mouse" X-GVHD model, and there was no significant difference between intraperitoneal infusion pathway and caudal vein infusion pathway in the establishment of "human mouse" X-GVHD model. The "human mouse" X-GVHD model could be successfully established by intraperitoneal infusion of human PBMC. The optimal time window for monitoring the phenotypic changes and functions of human T cells was 7-11 days. The average survival time of the model rats was 14.16 卤1.77 days. The innovation and significance of this study:. In this study, different mouse models were used to establish a human NOD / SCID xenograft-versus-host disease model. The conditions were optimized and a stable human-mouse X-GVHD model was established. The model provides experimental basis for the study of inducing human T cell tolerance. It can also be used to study drug sensitivity in vivo, predict and guide the selection and design of clinical treatment schemes, and explore radiotherapy, endocrine therapy and immunotherapy. Gene therapy and molecular targeting therapy.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R392

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