A molecular network regulating the pro-inflammatory phenotype of human memory T lymphocytes (ChIP-Seq)

Understanding the mechanisms that modulate T helper lymphocyte functions is crucial to decipher normal and pathogenic immune responses in humans. To identify molecular determinants influencing the pathogenicity of T cells, we separated ex vivo-isolated primary human memory T lymphocytes based on their ability to produce high levels of inflammatory cytokines. We found that the inflammatory, cytokine-producing phenotype of memory T lymphocytes was defined by a specific core gene signature and was mechanistically regulated by the constitutive activation of the NF-kB pathway and by the expression of the transcriptional repressor BHLHE40. BHLHE40 attenuated the expression of anti-inflammatory factors, including miR-146a, a negative regulator of NF-kB activation, and ZC3H12D, an RNase of the Regnase-1 family able to degrade inflammatory transcripts. Our data reveal a molecular network regulating the pro-inflammatory phenotype of human memory T lymphocytes, with the potential to contribute to disease. Overall design: Jurkat T cells were lentivirally transduced either with a construct harboring Bhlhe40 or an empty vector (pLvx-EF1a-FlagHA-IRES-ZsGreen backbone)

阐明调控辅助性T淋巴细胞功能的机制，对于解析人类正常与病理性免疫应答至关重要。为鉴定影响T细胞致病性的分子决定因素，我们依据体外分离的原代人记忆T淋巴细胞产生高水平炎性细胞因子的能力对其进行分选。研究发现，记忆T淋巴细胞的炎性细胞因子产生表型由特定核心基因特征所定义，其机制受核因子κB（NF-kB）通路的组成型激活，以及转录抑制因子BHLHE40的表达调控。BHLHE40可抑制多种抗炎因子的表达，其中包括核因子κB激活的负调控因子微小RNA-146a（miR-146a），以及能够降解炎性转录本的Regnase-1家族核糖核酸酶ZC3H12D。本研究揭示了调控人记忆T淋巴细胞促炎表型的分子网络，该网络或可参与疾病的发生发展。实验整体设计：将Jurkat T细胞以携带Bhlhe40的构建体或空载体（pLvx-EF1a-FlagHA-IRES-ZsGreen骨架载体）进行慢病毒转导。