摘要
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。
The study designed to observe the proliferation and survival of leukemia cells in vivo, and investigate the effect of linker for activated T cells （LAT） and LAT palmitoylation in T cell activation and signal transduction by establishing lymphocytic leukemia animal model with human Jurkat cell in node mice. We cultivated human Jurkat cell lines at first. According to the inoculation cell type, the Balb/c female mice were randomly divided into 4 groups, including 3 experimental groups： normal Jurkat group, normal LAT transfection group, mutation LAT transfection group, and 1 blank control group, with 6 mice in each group. After two consecutive days of intraperitoneal injection of cyclophosphamide, experimental mice were intravenously injected with the prepared Jurkat cell lines 5×10^6, while control group mice were intravenously injected with equivalent PBS solution. Then the general symptoms, weight and the number of leukocyte in peripheral blood of mice were observed. Dying mice were executed, and pathological tissues were taken for HE staining; FCM was applied to detect the positive rate of tumor cells. Data showed that after inoculation with tumor cells, experimental mice gradually appeared weight loss, hunchback, listlessness, WBC increase and survival time shortening. And tumor cell invasion could be found in the tissues of marrow and liver-spleen. FCM test results showed that the normal LAT transfection group had the highest rate of tumor cells. Taken together, we established an animal model of human Jurkat cells in nude mice successfully and further confirm that LAT palmitoylation could promote the proliferation and activation of T cell, while mutation of LAT palmitoylation site would block the activation signal delivery of T cell.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2016年第2期109-113,共5页
Immunological Journal
基金
国家自然科学基金(81273206)
