Bulk RNA-Seq to study the response of cDC2 and Axl+DC to HIV-1 infection. Bulk RNA-Seq to study the response of cDC2 and Axl+DC to HIV-1 infection

Axl+DC have been initially described through their ontogeny. Our lab reported for the first time an Axl+ DC functional specificity in the context of HV-1 capture, infection and in vitro transmission as a consequence of their constitutive expression of Siglec-1. However, Axl+DC immune responses to viral exposure remained to be studied. This study aim to decipher innate immune response of Axl+DC when exposed to HIV-1 in comparison to other DC subtypes. Overall design: cDC2 and Axl+DC cells were cultured with HIV-1 NL-AD8 complemented with Vpx for 24h. Cells cultured in mock medium were used as a control, as well as cells exposed to HIV-1 in the presence of azidothymidine (Azt) and neverapin (Nvp) to inhibit retrotranscription. Cells stimulated with TLR-L were also included .

AxL阳性树突状细胞（Axl+DC）的特征最初通过其个体发育过程被阐明。本课题组首次报道了Axl+DC在捕获、感染人类免疫缺陷病毒1型（HIV-1）以及体外传播过程中的功能特异性，该特性源于其组成性表达Siglec-1。然而，Axl+DC针对病毒暴露的免疫应答仍有待深入研究。本研究旨在解析Axl+DC在暴露于HIV-1时的固有免疫应答，并与其他树突状细胞亚型进行对比分析。
整体实验设计：将经典树突状细胞2型（cDC2）与Axl+DC与搭载Vpx蛋白的HIV-1 NL-AD8共培养24小时。以空白培养基培养的细胞作为空白对照，同时设置暴露于HIV-1但添加叠氮胸苷（Azt）与奈韦拉平（Nvp）以抑制逆转录过程的实验组作为对照。此外还纳入了经Toll样受体配体（TLR-L）刺激的实验组。