Transcriptomic analysis of TCF-7 cKO CD4 T cells

The transcription factor T Cell Factor-1 encoded by (TCF-7) is critical for T cell development. However, the role of TCF-7 on peripheral CD4 T cells mediated allo-immunity was not known. In this report using a clinically relevant model we presented novel finding of how TCF-7 regulates CD4 T cells functions, including CD4 T cells activation, proliferation, differentiation and impacts effector and central memory formation. We uncovered how TCF-7 regulates chemokine receptor which are essential for CD4 T cells migration to the site of inflammation. Our data uncovered how TCF-7 plays critical role in CD4 T cells survival, apoptosis. Using in vivo allo-immunity models and in-vitro studies we demonstrated how TCF-7 plays central role in CD4 T cells mediated damaged to organs like liver, skin and small intestine. We provided both molecular and biochemical and transcriptomic evidence how TCF-7 functionally regulates CD4 T cells mediated apoptosis and cell death, T cell mediated processes, pro-inflammatory and anti-inflammatory cytokines productions in both basal level and after allo-activation. These findings novel findings represent a stem forward to designing target specific approach for CD4 T cells mediated diseases while understand molecular mechanism the role of CD4 T cells in allo-immunity. We analyzed FACS purified CD8 T cells of 3 different TCF-7 cKO and WT mouse labeled this group of samples as pre-transplanted samples. For post-transplanted samples 1 x10^6 CD3 T Cells from TCF-7 cKO and WT mice were initially transplanted into lethally irradiated recipent Balb/c mouse and at day 7 post-transplant H2kb+ donor CD4 and CD8 T cells FACS sorted from the spleen of recipients into the Trizol.

由TCF-7编码的转录因子T细胞因子-1（T Cell Factor-1，TCF-1）在T细胞发育过程中发挥关键作用。然而，此前学界对TCF-7在外周CD4 T细胞介导的同种异体免疫中的作用尚不明确。本研究采用临床相关模型，揭示了TCF-1调控CD4 T细胞功能的全新机制，涵盖CD4 T细胞活化、增殖、分化，以及对效应记忆与中枢记忆形成的影响。我们阐明了TCF-1如何调控趋化因子受体——这类受体是CD4 T细胞迁移至炎症位点的关键分子。本研究数据揭示了TCF-7在CD4 T细胞存活与凋亡过程中的核心作用。通过体内同种异体免疫模型与体外实验，我们证实了TCF-7在CD4 T细胞介导的肝、皮肤、小肠等器官损伤中扮演核心调控角色。我们从分子、生化与转录组层面提供了证据，阐明TCF-7如何功能性调控CD4 T细胞介导的凋亡与细胞死亡、T细胞相关生物学过程，以及基础状态与同种异体活化后促炎与抗炎细胞因子的产生。这些全新发现为设计针对CD4 T细胞介导疾病的靶向特异性疗法迈出了重要一步，同时也加深了我们对CD4 T细胞在同种异体免疫中作用的分子机制的理解。我们分析了3组TCF-7条件性敲除（conditional knockout，cKO）与野生型（Wild Type，WT）小鼠的荧光激活细胞分选（Fluorescence Activated Cell Sorting，FACS）纯化CD8 T细胞，并将该组样本标记为移植前样本。对于移植后样本，我们将1×10^6个来自TCF-7 cKO与WT小鼠的CD3 T细胞移植至经致死剂量辐照的Balb/c受体小鼠体内，并在移植后第7天，从受体小鼠脾脏中FACS分选得到H2kb阳性供体CD4与CD8 T细胞，将其置于Trizol试剂中保存。