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目的建立人Jurkat淋巴细胞白血病裸鼠模型,观察Jurkat细胞在小鼠体内存活及增殖情况,研究T细胞活化连接蛋白(LAT)及其棕榈酰化在T细胞活化信号转导中的相关作用。方法将Balb/c纯系雌性裸鼠随机分为正常Jurkat组、正常LAT转染组、突变LAT转染组以及空白对照组,每组6只。连续2 d腹腔定量注射环磷酰胺后,实验组小鼠尾静脉注射人Jurkat细胞株5×106/只,对照组小鼠尾静脉注射等量PBS溶液。观察小鼠发病一般体征、体质量及外周血白细胞数量变化,濒死小鼠处死后取病理组织进行HE染色观察。流式细胞术(FCM)检测肿瘤细胞阳性率。结果接种肿瘤细胞后,实验组小鼠逐渐出现体质量减轻、弓背、精神萎靡等症状,外周血WBC计数逐渐增高,生存时间缩短。骨髓及肝脾组织可见肿瘤细胞浸润。流式细胞仪检测结果显示,正常LAT转染组肿瘤细胞阳性率高于其他各组。结论成功建立人Jurkat细胞裸鼠动物模型,从体内实验进一步证实LAT棕榈酰化促进T细胞活化增殖,而LAT棕榈酰化位点突变后,将阻碍T细胞活化信号的传递。
Objective To establish a human Jurkat lymphoblastic leukemia model in nude mice and to observe the survival and proliferation of Jurkat cells in mice and to investigate the role of T lymphocyte activation-associated protein (LAT) and palmitoylation in T cell activation signal transduction. Methods Balb / c pure female nude mice were randomly divided into normal Jurkat group, normal LAT transfection group, mutant LAT transfection group and blank control group, with 6 mice in each group. After continuous intraperitoneal injection of cyclophosphamide for 2 days, the mice in experimental group were injected with 5 × 106 human Jurkat cells via the tail vein, and the mice in the control group were injected with the same volume of PBS through the tail vein. Observe the general signs of onset of disease, body weight and peripheral blood leukocyte changes in morbidity mice were sacrificed after pathological HE staining. Flow cytometry (FCM) detection of tumor cell positive rate. Results After inoculation of tumor cells, mice in the experimental group gradually developed symptoms of reduced body mass, dorsal bowel, apathetic symptoms, increased WBC counts in peripheral blood and shortened survival time. Bone marrow and liver and spleen tissue visible tumor cell infiltration. The results of flow cytometry showed that the positive rate of tumor cells in normal LAT transfection group was higher than other groups. Conclusion The animal model of human Jurkat cells was established successfully. In vivo experiments further confirmed that LAT palmitoylation can promote the activation and proliferation of T cells. However, mutation of LAT palmitoylation site will block the signal transduction of T cell activation.