Profiling of CD4+ T cells responding to transient or persistent antigen presented by dendritic cells in vivo

These experiments were done to compare the gene expression profiles in CD4+ T cells responding to antigen presented by dendritic cells transiently or persistently. Some treatments include the activation of the dendritic cells by CD40 engagement. Keywords: immune response 2-4x106 lymph node cells from AND TCR-transgenic animals that expressed the congenic marker CD90.1 were transferred into double-transgenic (Ii-rTA/TIM) recipient animals that expressed the corresponding peptide from moth cytochrome c under a doxycycline-controllable promoter and were FACS-sorted 60 hours later. Recipients expressed the antigen not at all (“ctrl”), transiently (“short”) or throughout the experiment (“long”) due to different doxycycline treatment protocols. A second variable tested was the activation status of the dendritic cells: recipients were either injected by the stimulatory antibody FGK45.5 targeting CD40 (“FGK”) or PBS (“PBS”).

本实验旨在比较CD4+ T细胞在识别由瞬时呈递或持续性呈递抗原的树突状细胞（dendritic cells）所呈递的抗原后的基因表达谱差异。部分实验组设置了通过CD40结合激活树突状细胞的处理方案。
关键词：免疫应答
将表达同类系标记CD90.1的AND TCR转基因动物的2×10^6至4×10^6个淋巴结细胞，移植至双转基因（Ii-rTA/TIM）受体动物体内；该受体动物可在多西环素（doxycycline）可控启动子的调控下，表达天蛾细胞色素c的对应肽段。细胞移植60小时后，通过流式细胞术（FACS）进行分选。
根据不同的多西环素处理方案，受体动物的抗原表达状态分为三类：完全不表达抗原（"ctrl"）、瞬时表达抗原（"short"），以及整个实验期间持续表达抗原（"long"）。
本实验测试的第二个变量为树突状细胞的激活状态：向受体动物分别注射靶向CD40的刺激性抗体FGK45.5（"FGK"组）或磷酸盐缓冲液（PBS，"PBS"组）。