Alarmin S100A11 initiates a chemokine response to the human pathogen Toxoplasma gondii

Purpose: The goal of this study is to compare NGS-derived transcriptome profiling of human PBMC infected with T. gondii, PRU and RH strains, (RNA-seq) to not stimulated human PBMC transcriptome (RNA-seq). Methods: After 24h post infection PBMC and non-infected PBMC mRNA profiles were generated by deep sequencing. The sequence reads were analyzed within Galaxy platform: FastQC(*) program was used to generate quality control reports followed by employment of Trimmamatic (*) to trim “noisy” ends; TopHat mapped reads to hg38 human genome; Cuffdiff (*) was used to find significant changes in transcript expression between infected with T. gondii human PBMC and not infected PBMC. Results: RNA-seq data for genes with a fold change ≥2.5 and p value <0.05. Conclusions: human innate responses to T. gondii were primarily characterized by the induction of chemokine expression, including CCL2 Human PBMC mRNA profiles of infected cell with T. gondii and non infected cells were generated by deep sequencing,in quintuplicate

研究目的：本研究旨在对比感染刚地弓形虫（Toxoplasma gondii）PRU株与RH株的人外周血单个核细胞（Peripheral Blood Mononuclear Cell, PBMC）的下一代测序（Next-Generation Sequencing, NGS）转录组谱（RNA测序（RNA Sequencing, RNA-seq）），与未受刺激的人PBMC转录组（RNA-seq）数据。
实验方法：感染后24小时，通过深度测序获取感染组与未感染组人PBMC的mRNA表达谱。测序读段在Galaxy平台上开展分析：首先使用FastQC(*)生成质量控制报告，随后通过Trimmamatic(*)修剪测序读段的“噪声”末端；利用TopHat将读段比对至hg38人类参考基因组；使用Cuffdiff(*)筛选刚地弓形虫感染的人PBMC与未感染PBMC之间转录本表达存在显著差异的基因。
研究结果：本研究提供了差异倍数≥2.5且p值<0.05的基因对应的RNA-seq数据。
研究结论：人针对刚地弓形虫的固有免疫应答主要以趋化因子表达的诱导为核心特征，包括CCL2；本研究中感染刚地弓形虫的细胞与未感染细胞的人PBMC mRNA表达谱均通过五重复实验的深度测序生成。