Supporting data for: Caspase-8 expression in CD8+ T cells promotes pathogen restriction in the brain during Toxoplasma gondii infection (MERFISH naive)

Cell death is an integral restriction mechanism against intracellular pathogens. We have previously reported extensive cell death in the brain during infection with the intracellular parasite, Toxoplasma gondii. Here we focus on the role of caspase-8, a regulator of extrinsic apoptosis, during T. gondii infection. We find that Casp8-/-Ripk3-/- mice have increased brain parasite burden in comparison to controls and succumb to infection, despite the generation of robust immune responses. We observed that neurons, astrocytes, and CD8+ T cells had high rates of parasite interactions in Casp8-/-Ripk3-/- mice compared to WT mice. While Casp8 deficiency in neurons and astrocytes did not impact control of infection, deletion of Casp8 in CD8+ T cells led to impaired survival, increased parasite burden and direct infection of CD8+ T cells in the brain. We conclude that in addition to well-characterized effector functions, CD8+ T cells utilize caspase-8 to control T. gondii in the brain.This item contains data for MERFISH (Multiplexed Error-Robust Fluorescence In Situ Hybridization), a single-molecule imaging technique that allows for spatially mapping of single nuclear transcripts. This item contains data for a naive brain including raw data, cell clustering script, cell by gene identification, and detected transcripts.

细胞死亡是抵御细胞内病原体的不可或缺的限制机制。我们此前曾报道，在感染细胞内寄生虫刚地弓形虫（Toxoplasma gondii）期间，大脑中会发生广泛的细胞死亡。本研究聚焦于半胱天冬酶-8（caspase-8）——一种外源性细胞凋亡调控因子——在刚地弓形虫感染过程中的作用。我们发现，与对照组相比，半胱天冬酶-8敲除/受体相互作用蛋白激酶3双敲除（Casp8-/-Ripk3-/-）小鼠的脑部寄生虫负荷更高，尽管其能产生强劲的免疫应答，却仍死于感染。我们观察到，与野生型（WT）小鼠相比，半胱天冬酶-8敲除/受体相互作用蛋白激酶3双敲除小鼠的神经元、星形胶质细胞以及CD8+ T细胞与寄生虫的相互作用频率更高。尽管神经元和星形胶质细胞中的半胱天冬酶-8缺失并不影响感染控制，但CD8+ T细胞中的半胱天冬酶-8缺失会导致小鼠存活能力受损、脑部寄生虫负荷升高，且大脑内的CD8+ T细胞会被直接感染。综上，除了已被充分阐明的效应功能外，CD8+ T细胞还可通过半胱天冬酶-8来调控脑部的刚地弓形虫感染。本数据集包含MERFISH（多重误差鲁棒荧光原位杂交，Multiplexed Error-Robust Fluorescence In Situ Hybridization）相关数据，该技术是一种可实现单个核转录本空间定位的单分子成像技术。本数据集包含未致敏脑组织相关数据，具体涵盖原始数据、细胞聚类脚本、细胞-基因识别结果以及已检测到的转录本。