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RNA-seq analysis of human T CD4+ cells after their interaction with autologous antigen-specific vitamin D3-generated tolerogenic dendritic cells

ABSTRACT: The use of autologous tolerogenic dendritic cells (tolDC) has become a promising alternative for the treatment of autoimmune diseases. Among the different alternatives available, the use of vitamin D3 for the generation of tolDC (vitD3‑tolDC) constitutes one of the most robust approaches due to their immune regulatory properties, which are currently being tested in clinical trials. However, the mechanisms that vitD3‑tolDC are triggering for the induction of tolerance remain elusive. For this reason, we have performed a full transcriptomic characterization of T cells upon their interaction with autologous, antigen-specific vitD3-tolDC. Our results evidenced a strong down-modulation of genes involved in cell cycle and cell response to immune-related stimuli in T CD4+ cells after their interaction with vitD3-tolDC. Consequently, our results show the induction of a strong antigen-specific hyporresponsiveness, which manifests the regulatory properties of these cells and therefore their therapeutic potential in the clinic.

ORGANISM(S): Homo sapiens

PROVIDER: GSE128816 | GEO | 2020/12/21

REPOSITORIES: GEO

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RNA-seq analysis of human T CD4+ cells after their interaction with autologous antigen-specific vitamin D3-generated tolerogenic dendritic cells

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Project description:The present study reports an unbiased analysis of the genetic profile and regulation of NKG2D expressing CD4 T-cells.An Affymetrix microarray analysis was used to explore the genetic profile of NKG2D+ versus NKG2D- CD4 T-cells. The genetic profile was studied by single gene analysis and gene set enrichment analysis. I found that several immune regulatory receptors was regulated differently in NKG2D+ versus NKG2D- CD4 T-cells. Futhermore, I found that NKG2D+ CD4 T-cells display a genetic profile of cytotoxic T-cells. The gene set enrichment analysis revealed a change in 19 processes, including ARF GTPase activator activity; RNA splicing; Signal transduction; Interspecies interaction between organisms; Regulation of ARF GTPase activity; Cell motility; Mitosis; Cell cycle; Anaphase-promoting complex-dependent proteasomal ubiquitin-dependent protein catabolic process; Induction of apoptosis by extracellular signals; Negative regulation of apoptosis; mRNA export from nucleus; Positive regulation of ubiquitin-protein ligase activity involved in mitotic cell cycle; Cell division; Protein polymerization; Spliceosome assembly; Microtubule-based movement; Immune response; mRNA processing. Human CD4 T-cells were isolated from buffy coat by anti-CD4 conjugated magnetic beads. Memory CD4 T-cells were sorted, labeled with CFSE and stimulated with autologous monocytes pulsed with HCMV. HCMV stimulated CD4 T-cells were sorted as CFSE-NKG2D+ or CFSE-NKG2D- and processed for microarray analysis.

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