HIT analysis of T cells activated in the presence of TGF-β

We have developed a multi-analyte fluid-phase protein array technology termed high-throughput immunophenotyping using transcription (HIT). This method employs a panel of monoclonal antibodies, each tagged with a unique oligonucleotide sequence that serves as a molecular barcode. After staining a sample, T7 polymerase amplifies the tags which are then hybridized to a DNA microarray for indirect measurement of each analyte. Here we screened 90 antibodies directed against a panel of cell surface markers as well as 4 isotype controls to compare human naive CD4+ T cells activated in the presence or absence of TGF-β. Keywords: response to stimulus Two-condition experiment, with or without TGF-β. Biological replicates: 3 normal human donors, each donor served as an internal control. One replicate per array. One of the biological replicates was performed as a dye-swap to monitor dye-bias.