Single cell analysis of CD1a-restricted Group A streptococcus (GAS)-reactive peripheral and cutaneous T cells

Skin and blood single cell RNA sequencing datasets of IL-22- and IFNγ-secreting CD1a-reactive GAS-responsive T cells were generated from four healthy skins, five healthy PBMCs and three psoriatic PBMCs. Using cellular indexing of transcriptomes and epitopes by sequencing (CITE-seq), we measured T cell specific functional states at both protein and transcriptome levels. We deployed CD1a-expressing K562 cells as antigen presenting cells. K562-CD1a were infected with GAS, and were recognized by ex vivo cutaneous and blood T cells in a CD1a-dependent manner leading to production of IL-22 or IFNγ, detected using Cytokine Secretion assays. FACS-isolated CD3+ T cells with or without prior K562 co-culture were included to establish the dataset. CITE-seq antibodies against the fluorochromes PE or APC on the detection antibodies were included to further characterise cytokine-producing T cells.

本数据集针对分泌白介素22（IL-22）与干扰素γ（IFNγ）的CD1a反应性A组链球菌（Group A Streptococcus，GAS）应答性T细胞，从4份健康皮肤样本、5份健康外周血单个核细胞（Peripheral Blood Mononuclear Cell，PBMC）样本及3份银屑病患者外周血单个核细胞样本中，构建了皮肤与血液来源的单细胞RNA测序（single cell RNA sequencing）数据集。本研究采用测序转录组与表位的细胞索引技术（Cellular Indexing of Transcriptomes and Epitopes by Sequencing，CITE-seq），在蛋白质与转录组两个层面同步检测T细胞的特异性功能状态。以表达CD1a的K562细胞作为抗原呈递细胞（antigen presenting cells），将K562-CD1a细胞感染A组链球菌后，其可通过CD1a依赖的方式被离体皮肤与血液T细胞识别，进而诱导IL-22或IFNγ的分泌，该过程通过细胞因子分泌检测实验得以验证。为完成本数据集的构建，纳入了经荧光激活细胞分选（Fluorescence-Activated Cell Sorting，FACS）分离的CD3阳性T细胞，包含预先与K562细胞共培养及未共培养的两组样本。此外，本研究使用针对检测抗体上藻红蛋白（Phycoerythrin，PE）与别藻蓝蛋白（Allophycocyanin，APC）荧光基团的CITE-seq抗体，进一步对分泌细胞因子的T细胞进行表型鉴定。