輸注人PBMC建立“人-鼠”異種移植物抗宿主病模型1
本文選題:PBMC + 器官移植; 參考:《中國免疫學(xué)雜志》2014年12期
【摘要】:目的:摸索X-GVHD模型的合適小鼠品系,建立穩(wěn)定的"人-鼠"X-GVHD模型。方法: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(xì)胞(PBMC)建立異種X-GVHD模型。通過檢測人T細(xì)胞在模型鼠外周血、組織、器官中的浸潤情況等指標(biāo)(采用流式細(xì)胞術(shù)和免疫組化技術(shù)),比較兩種模型鼠中人免疫細(xì)胞的浸潤率和各組生存時間,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(xì)胞量,以及觀察免疫細(xì)胞表型變化與功能的最佳時間窗口。結(jié)果: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(xì)胞表型動態(tài)變化與功能的最佳時間窗口為7~11 d;模型鼠的平均生存時間為(14.16±1.77)d。結(jié)論:NOD/SCID鼠通過腹腔注射5×107以上人PBMC可以成功建立"人-鼠"X-GVHD模型,7~11 d為監(jiān)測人T細(xì)胞表型動態(tài)變化與功能的最佳時間。
[Abstract]:Objective: to explore the suitable strain of X-GVHD model and to establish a stable human-mouse X-GVHD model. Methods: the xenogeneic X-GVHD model was established in nude mice after whole body irradiation with sublethal dose 緯-ray and intraperitoneal infusion of PBMCs from peripheral blood monocytes of healthy people. The infiltration rate and survival time of human T cells in peripheral blood, tissues and organs of model rats were compared by flow cytometry and immunohistochemical technique. Thus, the appropriate strain of X-GVHD model was established, and the infusion pathway and appropriate cell quantity of human PBMC were explored, and the best time window for observing the phenotypic changes and functions of immune cells was also found. Results the mice with "human-mouse" X-GVHD model were more suitable to be induced by the mice than the control group, and there was no significant difference between the intraperitoneal infusion pathway and the caudal vein infusion pathway in the establishment of the "human-mouse" X-GVHD model, and the "human-mouse" X-GVHD model could be successfully established by intraperitoneal infusion of more than 5 脳 10 ~ 7 human PBMC. The optimal time window for monitoring the phenotypic changes and functions of human T cells was 711 days, and the average survival time of model mice was 14.16 鹵1.77 days. Conclusion the "human-mouse" X-GVHD model can be successfully established by intraperitoneal injection of 5 脳 10 ~ (7) human PBMC into the control group. The best time to monitor the dynamic changes and function of human T cell phenotypes is to establish the "human-mouse" X-GVHD model for 7 ~ 11 days.
【作者單位】: 湖北醫(yī)藥學(xué)院附屬東風(fēng)醫(yī)院;武漢科技大學(xué)醫(yī)學(xué)院;華中科技大學(xué)同濟(jì)醫(yī)學(xué)院免疫學(xué)系;
【基金】:國家自然科學(xué)基金資助項目(30972697;30901320)
【分類號】:R392
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