Unknown,Transcriptomics,Genomics,Proteomics

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A combined omics approach to generate the surface protein atlas of human naive CD4+ T cells during early TCR activaion

ABSTRACT: To increase the dataset of our proteomic naive CD4+ T cell surface atlas, we applied whole genome microarray expression analysis. The NCBI RefSeq accession number of all transcripts which were detected in naive and stimulated CD4+ T cells (aCD3/aCD28 for 3h) of four different donors, was mapped to the corresponding UniProtKB accession number. For these UniProtKB ACs, we extracted the subcellular localization (UniProt_SL) annotation from the UniProtKB/Swiss-Prot database or if not available the subcellular localization was predicted unsing LocTree3 and PolyPhobius. This led to the identification of 908 genes coding for proteins located on the surface of human naive and/or activated CD4+ T cells. Gene expression in human naive T cells and naive T cells which were stimulated with aCD3/aCD28 for 3h was measured. Naive T cells from four different human blood donors were analyzed.

ORGANISM(S): Homo sapiens

SUBMITTER: Kathrin Suttner

PROVIDER: E-GEOD-61983 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Naive CD4+ T cells are the common precursors of multiple effector and memory T cell subsets and possess a high plasticity in terms of differentiation potential. This stem-cell like character is important for cell therapies aiming at regeneration of specific immunity. Cell surface proteins are crucial for recognition and response to signals mediated by other cells or environmental changes. Knowledge of cell surface proteins of human naive CD4+ T cells and their changes during the early phase of T cell activation is urgently needed for a guided differentiation of naive T cells and may support the selection of pluripotent cells for cell therapy.<br>Periodate oxidation and aniline-catalyzed oxime ligation (PAL) technology was applied with subsequent quantitative LC-MS/MS (PAL-qLC-MS/MS) to generate a dataset describing the surface proteome of human naive CD4+ T cells and to monitor dynamic changes during the early phase of activation. This led to the identification of 173 N-glycosylated surface proteins, of which 24 were previously not known to be expressed on human naive CD4+ T cells or have no defined role within T cell activation. To independently confirm the proteomic dataset and to analyse the cell surface by an alternative technique a systematic phenotypic expression analysis of surface antigens via flow cytometry was performed. This screening expanded the previous dataset, resulting in 229 surface proteins which are expressed on naive unstimulated and activated CD4+ T cells. Furthermore, we generated a surface expression atlas based on transcriptome data, experimental annotation and predicted subcellular localization, and correlated the proteomics result with this transcriptional dataset.<br>This extensive surface atlas provides an overall naive CD4+ T cell surface resource and will enable future studies aiming at a deeper understanding of mechanisms of T cell biology allowing the identification of novel immune targets usable for the development of therapeutic treatments.

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A combined omics approach to generate the surface protein atlas of human naive CD4+ T cells during early TCR activaion

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