Profiling the peripheral immune response to TNF-alpha stimulation in patients with untreated juvenile idiopathic arthritis using scRNAseq. Profiling the peripheral immune response to TNF-alpha stimulation in patients with untreated juvenile idiopathic arthritis using scRNAseq

We report the application of next-generation sequencing tehcnologies to profile the effects of tumor necrosis factor alpha (TNFα) treatment on the transcriptomes of cells derived from juvenile idopathic arthritis (JIA) patients and healthy control subjects. Whole blood from de-identified subects with JIA was collected following children's provided assent and parental signed consent. Controls included healthy adults under the age of 25 years without any medical conditions or family history of autoimmune disease. All study procedures were approved by the Emory IRB. Blood samples from each subject was then separated into two aliquots, with one supplemented with TNFα. After 24hr ex-vivo stimulation, PBMCs were isolated and subject to cDNA library sequencing on a NextSeq 550. Overall design: Peripheral blood single-cell mRNA profiles for 6 JIA patients and 2 healthy controls, with each subject having a corresponding unstimulated and a 24 TNFα ex-vivo stimulated sample.

本研究报道了下一代测序技术（next-generation sequencing）的应用，用于解析肿瘤坏死因子α（tumor necrosis factor alpha, TNFα）处理对幼年特发性关节炎（juvenile idiopathic arthritis, JIA）患者及健康对照受试者来源细胞转录组的影响。收集去标识化的JIA患者全血样本前，已获得儿童受试者的知情同意及家长签署的书面知情同意书。健康对照纳入标准为年龄25岁以下、无任何基础疾病及自身免疫性疾病家族史的健康成人。本研究所有实验流程均已通过埃默里大学机构审查委员会（Institutional Review Board, IRB）批准。随后将每位受试者的血液样本分为两份等分试样，其中一份添加TNFα进行处理。经24小时离体刺激后，分离外周血单个核细胞（Peripheral Blood Mononuclear Cells, PBMCs），并在NextSeq 550测序平台上开展cDNA文库测序。实验设计概述：本研究包含6名JIA患者与2名健康对照的外周血单细胞mRNA测序数据，每位受试者均对应未刺激组与经24小时离体TNFα刺激组的配对样本。