Abstract：
Follicular Helper T (TFH) cells are perceived as an independent CD4+ T cell lineage that assists cognate B cells in producing high-affinity antibodies, thus establishing long-term humoral immunity. During acute viral infection, the fate commitment of virus-specific TFH cells is determined in the early infection phase, and investigations of the early-differentiated TFH cells are crucial in understanding T cell-dependent humoral immunity and optimizing vaccine design. In the study, using a mouse model of acute lymphocytic choriomeningitis virus (LCMV) infection and the TCR-transgenic SMARTA (SM) mouse with CD4+ T cells specifically recognizing LCMV glycoprotein epitope I-AbGP66-77, we described procedures to access the early fate commitment of virus-specific TFH cells based on flow cytometry stainings. Furthermore, by exploiting retroviral transduction of SM CD4+ T cells, methods to manipulate gene expression in early-differentiated virus-specific TFH cells are also provided. Hence, these methods will help in studies exploring the mechanism(s) underlying the early commitment of virus-specific TFH cells.
摘要：
卵泡辅助T（TFH）细胞被认为是一个独立的CD4+T细胞谱系，协助同源B细胞产生高亲和力抗体，从而建立长期的体液免疫。在急性病毒感染期间，病毒特异性TFH细胞的命运承诺是在感染早期确定的，早期分化的TFH细胞的研究对于理解T细胞依赖性体液免疫和优化疫苗设计至关重要。在研究中,使用急性淋巴细胞脉络膜脑膜炎病毒(LCMV)感染小鼠模型和具有特异性识别LCMV糖蛋白表位I-AbGP66-77的CD4+T细胞的TCR转基因SMARTA(SM)小鼠,我们描述了基于流式细胞术获得病毒特异性TFH细胞早期命运承诺的程序.此外，通过利用SMCD4+T细胞的逆转录病毒转导，还提供了在早期分化的病毒特异性TFH细胞中操纵基因表达的方法。因此，这些方法将有助于研究探索病毒特异性TFH细胞早期定型的潜在机制.
