Project description:Purpose: To elucidate the potential immune-regulatory mechanism of TRAIL of activated T cells in EAE, we analyzed gene expression profiles of splenic CD4+ T cells from EAE mice treated with TRAIL by RNA sequencing and transcriptome analysis. Methods: Splenic CD4+ T cell mRNA profiles of control or EAE mice treated with vehicle or TRAIL were generated by deep sequencing using Illumina Solexa. Library construction of all samples were used by Agilent's SureSelect Strand Specific RNA Library Preparation Kit for 75SE (Single-End or Paired-End) sequencing on Solexa platform. The sequence was directly determined using sequencing-by-synthesis technology via the TruSeq SBS Kit. Raw sequences were obtained from the Illumina Pipeline software bcl2fastq v2.0 and expected to generate 20M (million reads or Gb) per sample. Results: By comparing mRNA exression of splenic CD4+ T cells from EAE mice treated with vehicle or TRAIL, there were 244 genes significantly differentially expressed. By analyzing these 244 significant genes categorized by a Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, the most significantly enriched category for CD4+ T cells was “cell cycle” (multiple of enrichment: 3.13, p = 1.64 × 10−5) followed by “TCR signaling pathway” (multiple of enrichment: 2.80, p = 8.25 × 10−4). In addition, significant genes in the“TCR signaling pathway” tended to be downregulated while those in “cell cycle”tended to be upregulated in a volcano plot analysis. Consistent with the volcano plot results, by using unsupervised hierarchical clustering, the heatmap showed that significant TCR signaling pathway-associated genes were downregulated, while significant cell cycle-associated genes were upregulated in splenic CD4+ T cells from EAE mice treated with TRAIL Conclusions: Our study demonstrated the gene transcription pattern from CD4+ T cells of TRAIL-treated EAE mice were involved in distinct TCR signaling and cell cycle pathways.
Project description:EAE is a mouse model of human multiple sclerosis. We used miRNA low density array to screen abnormally expressed miRNAs in autoimmune CD4+T cells.Generally, splenic CD4+T cells were isolated from three groups: healthy C57BL/6 mice, C57BL/6 mice with the induction of EAE for 16 days, and C57BL/6 mice with the induction of EAE for 32 days. Then we did miRNA profilings on these samples. Splenic CD4+T cells were isolated from the above three groups. Each group consist of six mice. Equal amount total RNA from each mouse was pooled prior to miRNA expression analysis.
Project description:EAE is a mouse model of human multiple sclerosis. We used miRNA low density array to screen abnormally expressed miRNAs in autoimmune CD4+T cells.Generally, splenic CD4+T cells were isolated from three groups: healthy C57BL/6 mice, C57BL/6 mice with the induction of EAE for 16 days, and C57BL/6 mice with the induction of EAE for 32 days. Then we did miRNA profilings on these samples.
Project description:We isolated CD4 T cells from EAE mice with or without TRAIL treatment and analyzed by RNA-seq. We also activated naïve CD4 T cells by anti-CD3/CD28 antibodies in the presence or absence of TRAIL and subsequently analyzed using RNA-seq.
Project description:To investigate whether the level of O-GlcNAc could affect CD4+ T cell function after EAE, we performed RNA-sequencing analysis on splenic CD4+ T cells. The downregulation of OGT in CD4+ T cells led to changes in gene expression profile in CD4+ T cells with a 2-fold threshold. Next, Gene Ontology (GO) analysis was performed to compare differentially expressed genes in Ogt-KD and control mice. These results indicated that OGT was involved in the regulation of multiple important cellular events.
Project description:Investigation of the change of the Trail-dependent NK cell transcriptome during short-term (24h) infection with lymphocytic choriomeningitis virus (LCMV). RNA sequencing-based transcriptomics analysis was performed in spleen-isolated (NK1.1+CD3-) NK cells from 3 naïve Trail+/+ mice, 3 naive Trail-/- mice, 4 LCMV-infected Trail+/+ mice, and 4 LCMV-infected Trail-/- mice.
Project description:Assessment of whether endogenous IFN-I exerts genome-wide gene expression changes in splenic CD3 T cells and spinal cord during priming phase of EAE, we performed transcriptome analysis on T cells and spinal cord derived from 3-month old IFNAR1Texcl and WT animals on day 10 upon EAE induction.
Project description:Purpose: To compare gene expression of CD4+ T cells between Type-A and Type-B EAE Method: CD4+ T cells were FACS sorted from spleens 9-day post immunization. Results: We found that Type-B EAE (NLRP3 inflammasome-dependent and IFNb treatment-resistant EAE) express high levels of CXCR2 and other chemokine receptors, which can be targeted to treat the Type-A EAE subtypes, instead of treating with IFNb. Conclusions: Altering intensity of immunization significantly impacts on gene expression patterns in T cells.