Gene expression profile at single-cell level of helped and helpless LCMV-specific CD8+ effector T cells (day 7 post acute LCMV infection)

We used single-cell RNA sequencing (scRNA-seq) to obtain transcriptomic profiles of virus-specific CD8+ effector T cells generated in the presence (helped) or absence (helpless) of CD4+ T cells. Young adult B6 mice were treated with isotype control antibody (helped) or depleted of CD4+ T cells (helpless), transferred intravenuously with 5,000 naïve congenic (CD90.1+) T cell receptor transgenic (TCRtg) CD8+ T cells specific for the LCMV GP33-41 determinant (P14 cells), and challenged with LCMV Armstrong (2e5 pfu intraperitoneally). Seven days later, live P14 effector T cells (P14 TE) were purified from spleens of P14 chimeras by fluorescence activated cell sorting for downstream scRNA-seq processing.

本研究采用单细胞RNA测序（single-cell RNA sequencing, scRNA-seq），获取了两类病毒特异性CD8+效应T细胞的转录组图谱：一类为CD4+ T细胞存在条件下诱导生成的获辅助组（helped）细胞，另一类为CD4+ T细胞缺失条件下诱导生成的无辅助组（helpless）细胞。实验中，我们选取年轻成年B6小鼠分为两组：一组予以同型对照抗体处理，对应获辅助组（helped）；另一组进行CD4+ T细胞耗竭处理，对应无辅助组（helpless）。随后经静脉输注5000个未致敏的同源（CD90.1+）T细胞受体转基因（T cell receptor transgenic, TCRtg）CD8+ T细胞——该细胞为针对淋巴细胞脉络丛脑膜炎病毒（Lymphocytic Choriomeningitis Virus, LCMV）GP33-41表位的特异性T细胞，又称P14细胞；继而以2×10^5 噬斑形成单位（plaque-forming unit, pfu）的LCMV Armstrong毒株经腹腔注射进行感染攻击。造模7天后，通过荧光激活细胞分选术从P14嵌合小鼠的脾脏中纯化得到存活的P14效应T细胞（P14 TE），用于后续的单细胞RNA测序实验流程。