Genomic-Scale Transcriptional Analysis of T-Cell Activation Reveals Novel Genes and Signaling Programs

Min Wang

doi:https://doi.org/10.21985/N27B1H

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Genomic-Scale Transcriptional Analysis of T-Cell Activation Reveals Novel Genes and Signaling Programs

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T-cell activation is an essential step of the immune response, yet the cellular and molecular events underlying this complex process are not fully understood. Significantly, a comparative genome-scale transcriptional analysis of two T-cell subsets and the natural-mixed CD3+ population remains unexplored. Using microarrays, we investigated the temporal global transcriptional profile of human T-cell activation in CD3+, CD4+ and CD8+ T cells. Comparison of the microarray-based gene expression patterns between the three populations revealed largely conserved transcriptional patterns. Transcription patterns of selected genes were confirmed by Q-RT-PCR. We employed a Gene-Ontology-driven transcriptional analysis coupled with protein-abundance/activity assays to identify novel genes and cell-type-specific genes involved in the activation process. Focusing on immune response, we identified potential genes involved in the communication between the two subsets, would-be effector-function-specific genes and novel chemokines in T cells. Increased expression of unexpected cytokines (GPI, OSM and MIF) suggests their involvement in T-cell activation. Differential expression of many receptors, novel in T-cell activation, including CCR5, TNFRSF25 and TNFRSF1A, suggests their role in this process. >We identified significantly regulated apoptotic genes in several protein families and detailed their transcriptional kinetics during the T-cell activation process. Transcription patterns of some selected genes were validated at the protein level. The simultaneous upregulation of NF-κB and IκB family genes at 48-96 hours, supported by the increase of phosphorylated p65 at 48-96 hours, suggests the NF-κB involvement. Examination of significant regulated genes revealed an increase of p38 and ERK1 signalings during T-cell proliferation (48-96 hours), explored using phosphorylation assays for p38 and ERK1.. Gene-Ontology-driven transcriptional analysis of T-cell response to H2O2 stress revealed transcription events, such as response to stimulus, and significant genes, such as BBC3, involved in this stress response process in the context of T-cell activation. Analysis of T-cell activation, which integrates dynamic gene-expression data with protein-abundance and activity assays, has identified numerous novel genes and pathways that may be important players in T-cell activation. This has significantly broadened our understanding of the molecular orchestration of T-cell activation process, and provides a basis for further studies for understanding T-cell activation in health but also in malignancies and autoimmunity.

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