本文選題：器官移植　切入點：異種移植　出處：《華中科技大學(xué)》2011年碩士論文

【摘要】：在人體內(nèi)的實驗由于道德、倫理、法律等諸方面的制約而受到嚴格限制,涉及人體細胞的研究一般只能進行體外實驗,而體外實驗的結(jié)果往往與體內(nèi)情況存在一定的差別;“人-鼠”模型的建立,為在體內(nèi)研究提供了可能。然而目前對人-鼠模型的報道也較多,但行之有效且較穩(wěn)定的方法報道甚少�？晒┙梃b的異種移植物抗宿主病模型(graft-versus-host disease,GVHD)以“人-鼠”模型應(yīng)用最廣泛,但建立的方法也有較多的報道。本研究主要探討選擇合適小鼠品系,優(yōu)化相關(guān)實驗條件,建立穩(wěn)定的“人-鼠”異種移植物抗宿主病(X-GVHD)模型,為研究誘導(dǎo)人T細胞耐受提供動物模型。主要內(nèi)容與結(jié)果如下: 本研究選取了裸鼠、NOD/SCID經(jīng)亞致死劑量γ射線全身照射后,腹腔輸注健康人外周血單核細胞(PBMC)建立異種急性X-GVHD模型。通過檢測人T細胞在尾靜脈血、組織、器官中的浸潤情況等指標(采用流式細胞術(shù)和免疫組化技術(shù)),比較兩種模型鼠中人免疫細胞的浸潤率,各組生存時間,從而確定建立X-GVHD模型的合適小鼠品系,并摸索人PBMC的輸注途徑和合適細胞量,以及觀察免疫細胞表型變化與功能的最佳時間窗口。 結(jié)果顯示,NOD/SCID鼠更適合誘導(dǎo)“人鼠”X-GVHD模型的小鼠;腹腔輸注途徑和尾靜脈輸注途徑對建立“人鼠”X-GVHD模型無明顯差異;腹腔輸注5×107以上人PBMC可以成功建立“人鼠”X-GVHD模型。在采用優(yōu)化后的實驗條件建立模型中,監(jiān)測人T細胞表型動態(tài)變化與功能的最佳時間窗口為7-11天;模型鼠的平均生存時間為14.16±1.77天。 本研究的創(chuàng)新點和意義: 本研究使用不同的小鼠建立模型,經(jīng)檢測后,建立了“人-NOD/SCID異種急性移植物抗宿主病模型”,進一步優(yōu)化各項條件,建立了穩(wěn)定的“人-鼠”X-GVHD模型,該模型為誘導(dǎo)人T細胞耐受的研究提供了實驗依據(jù)。同時該模型可以用于體內(nèi)藥敏實驗研究,預(yù)測指導(dǎo)臨床治療方案的選擇和設(shè)計,以及探索放療、內(nèi)分泌治療、免疫治療、基因治療及分子靶向治療等新型的治療手段。
[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 緯 -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 鹵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.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2011
【分類號】：R392

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