生长方案:	CCR6+ memory CD4+ T cells from HLA-DR4+ healthy controls and HLA-DR4+ MS patients were amplified by PHA and IL-2 and stimulated by irradiated autologous monocytes and DR4 myelin peptides (MOG97-109 and PLP180-199). Cell proliferation was measured and two of the highest proliferated wells were picked for DR4 tetramers staining (MOG97-109-tetramers and PLP180-199-tetramers). Myelin tetramer+ and tetramer− cells were sorted into RNA lysis buffer for RNA sequencing.
处理方案:	Amplified T cell libraries assay were carried out as previous described (Geiger, 2009). Naïve, CCR6− and CCR6+ memory CD4+ T from paired MS patients and healthy controls were pre-sorted and cultured in 96-well round-bottom plates (Costar) at 2 × 103 cells per well in complete RPMI 1640 medium, and stimulated with 1 μg/ml PHA (Roche) and 20 U/ml IL-2 in the presence of irradiated (45Gy) allogeneic feeder cells (2×104/well). IL-2 was added on day 4, 7 and 10. Cultures were washed and split into two 96-well plates after 2-weeks of stimulation and expansion. Library screening was performed by culturing ~ 106 T cells/well with autologous monocytes (~105), which were either unpulsed or pulsed for 3 h with 10 μg/ml myelin peptide pools (MBP85-99, MOG222-241, PLP30-49, PLP129-148, MOG97-109, PLP180-199), or C. albicans (GREER). [3H] (Perkin Elmer) was added into the cultures 16 h before harvest. On day 5, cell proliferation was measured by 3H-thymidine incorporation on a scintillation beta-counter (Perkin Elmer). Culture supernatants were taken on day 7 for cytokine profiling as described below.
提取方案:	RNA was extracted using the NucleoSpin RNA XS Kit (Macherey-Nagel, Bethlehem, PA) according to the manufacturers instructions.
建库方案:	cDNA synthesis and amplification were performed using SMARTer Ultra Low Input RNA for Illumina Sequencing High Volume Kit (Clontech, Mountain View, CA) according to the manufacturers instructions. The average number of cells used to isolate RNA was 3,000, which yielded >1ng input RNA. Paired-end sequencing libraries were prepared using the Nextera XT DNA sample Prep Kit (Illumina, SanDiego, CA) according to the manufacturers instructions.

分子类型:	rRNA- RNA
库的片段类型:	PAIRED
库的链类型:	-
测序平台:	ILLUMINA
测序仪型号:	Illumina HiSeq 2500
链特异性:	Unspecific

数据来源	GEN样本编号	GEN数据集编号	系列编号	项目编号	样本编号	样本名称	生物样本编号	样本访问号	实验访问号	释放时间	提交时间	最后更新时间	物种	种族	族裔	年龄	年龄单位	性别	来源名称	组织	细胞类型	细胞亚型	细胞系	疾病	疾病状态	发育阶段	突变/变异	表型	Condition Detail	生长方案	处理方案	提取方案	建库方案	分子类型	库的片段类型	链特异性	库的链类型	加标（Spike-In）	测序方法	测序平台	测序仪型号	细胞数	测序片段数	碱基数	平均测序片段长度_1	平均测序片段长度_2	唯一比对率	多重比对率	覆盖度