PDF(Pubmed)
Abstract：
Individual T cell responses vary significantly based on the microenvironment present at the time of immune response and on prior induced T cell memory. While the cecal ligation and puncture (CLP) model is the most commonly used murine sepsis model, the contribution of diverse T cell responses has not been explored. We defined T cell subset responses to CLP using single-cell RNA sequencing and examined the effects of prior induced T cell memory (Immune Education) on these responses. We hypothesized that Immune Education prior to CLP would alter T cell responses at the single cell level at a single, early post-CLP time point.
Splenic T cells were isolated from C57BL/6 mice. Four cohorts were studied: Control, Immune-Educated, CLP, and Immune-Educated CLP. At age 8 weeks, Immune-Educated and Immune-Educated CLP mice received anti-CD3ϵ antibody; Control and CLP mice were administered an isotype control. CLP (two punctures with a 22-gauge needle) was performed at 12-13 weeks of life. Mice were sacrificed at baseline or 24-hours post-CLP. Unsupervised clustering of the transcriptome library identified six distinct T cell subsets: quiescent naïve CD4+, primed naïve CD4+, memory CD4+, naïve CD8+, activated CD8+, and CD8+ cytotoxic T cell subsets. T cell subset specific gene set enrichment analysis and Hurdle analysis for differentially expressed genes (DEGs) were performed.
T cell responses to CLP were not uniform - subsets of activated and suppressed T cells were identified. Immune Education augmented specific T cell subsets and led to genomic signatures favoring T cell survival in unoperated and CLP mice. Additionally, the combination of Immune Education and CLP effected the expression of genes related to T cell activity in ways that differed from CLP alone. Validating our finding that IL7R pathway markers were upregulated in Immune-Educated CLP mice, we found that Immune Education increased T cell surface IL7R expression in post-CLP mice.
Immune Education enhanced the expression of genes associated with T cell survival in unoperated and CLP mice. Induction of memory T cell compartments via Immune Education combined with CLP may increase the model\'s concordance to human sepsis.
Splenic T cells were isolated from C57BL/6 mice. Four cohorts were studied: Control, Immune-Educated, CLP, and Immune-Educated CLP. At age 8 weeks, Immune-Educated and Immune-Educated CLP mice received anti-CD3ϵ antibody; Control and CLP mice were administered an isotype control. CLP (two punctures with a 22-gauge needle) was performed at 12-13 weeks of life. Mice were sacrificed at baseline or 24-hours post-CLP. Unsupervised clustering of the transcriptome library identified six distinct T cell subsets: quiescent naïve CD4+, primed naïve CD4+, memory CD4+, naïve CD8+, activated CD8+, and CD8+ cytotoxic T cell subsets. T cell subset specific gene set enrichment analysis and Hurdle analysis for differentially expressed genes (DEGs) were performed.
T cell responses to CLP were not uniform - subsets of activated and suppressed T cells were identified. Immune Education augmented specific T cell subsets and led to genomic signatures favoring T cell survival in unoperated and CLP mice. Additionally, the combination of Immune Education and CLP effected the expression of genes related to T cell activity in ways that differed from CLP alone. Validating our finding that IL7R pathway markers were upregulated in Immune-Educated CLP mice, we found that Immune Education increased T cell surface IL7R expression in post-CLP mice.
Immune Education enhanced the expression of genes associated with T cell survival in unoperated and CLP mice. Induction of memory T cell compartments via Immune Education combined with CLP may increase the model\'s concordance to human sepsis.
摘要：
个体T细胞应答基于免疫应答时存在的微环境和先前诱导的T细胞记忆而显著变化。虽然盲肠结扎和穿孔（CLP）模型是最常用的小鼠败血症模型，不同T细胞反应的贡献尚未被探索。我们使用单细胞RNA测序定义了对CLP的T细胞亚群反应，并检查了先前诱导的T细胞记忆（免疫教育）对这些反应的影响。我们假设在CLP之前的免疫教育会在单个细胞水平上改变T细胞反应，CLP后的早期时间点。
■从C57BL/6小鼠中分离脾T细胞。研究了四个队列：对照，免疫教育，CLP,和免疫教育CLP。在8周大的时候，免疫教育和免疫教育的CLP小鼠接受抗CD3抗体;对照和CLP小鼠施用同种型对照。CLP（用22号针穿刺两次）在12-13周时进行。在基线或CLP后24小时处死小鼠。转录组文库的无监督聚类确定了六个不同的T细胞亚群：静止的幼稚CD4+，初免CD4+，记忆CD4+，幼稚CD8+，激活的CD8+，和CD8+细胞毒性T细胞亚群。进行T细胞亚群特异性基因集富集分析和差异表达基因(DEGs)的跨栏分析。
■T细胞对CLP的反应不一致-鉴定了活化和抑制的T细胞的亚群。免疫教育增强了特定的T细胞亚群，并导致基因组特征有利于未手术和CLP小鼠中的T细胞存活。此外,免疫教育和CLP的组合以不同于单独的CLP的方式影响与T细胞活性相关的基因的表达。验证我们的发现，IL7R途径标记在免疫教育的CLP小鼠中上调，我们发现免疫教育增加了CLP后小鼠T细胞表面IL7R的表达。
■免疫教育增强了未手术和CLP小鼠中与T细胞存活相关的基因的表达。通过免疫教育联合CLP诱导记忆T细胞区室可能会增加模型与人败血症的一致性。
■从C57BL/6小鼠中分离脾T细胞。研究了四个队列：对照，免疫教育，CLP,和免疫教育CLP。在8周大的时候，免疫教育和免疫教育的CLP小鼠接受抗CD3抗体;对照和CLP小鼠施用同种型对照。CLP（用22号针穿刺两次）在12-13周时进行。在基线或CLP后24小时处死小鼠。转录组文库的无监督聚类确定了六个不同的T细胞亚群：静止的幼稚CD4+，初免CD4+，记忆CD4+，幼稚CD8+，激活的CD8+，和CD8+细胞毒性T细胞亚群。进行T细胞亚群特异性基因集富集分析和差异表达基因(DEGs)的跨栏分析。
■T细胞对CLP的反应不一致-鉴定了活化和抑制的T细胞的亚群。免疫教育增强了特定的T细胞亚群，并导致基因组特征有利于未手术和CLP小鼠中的T细胞存活。此外,免疫教育和CLP的组合以不同于单独的CLP的方式影响与T细胞活性相关的基因的表达。验证我们的发现，IL7R途径标记在免疫教育的CLP小鼠中上调，我们发现免疫教育增加了CLP后小鼠T细胞表面IL7R的表达。
■免疫教育增强了未手术和CLP小鼠中与T细胞存活相关的基因的表达。通过免疫教育联合CLP诱导记忆T细胞区室可能会增加模型与人败血症的一致性。
