Expression data from mouse T cells after CLIP (Crosslinking immunoprecipitation) procedures (microRNA)

Molecular networks based on genome-wide mRNA and miRNA expression in activated T cells are integrated by co-regulation with miRNA and protein-coding genes during immune responses, however, accurate prediction of miRs and their targets remains a challenge. Furthermore, the specific changes in the expression of miRNAs and/or mRNAs in allogeneic activated T cells during HCT would be distinct from those responding to non-specific stimulation. Recently, Ago-CLIP has been demonstrated to provide a robust platform for identification of precise miRNA-mRNA interactions by definitively distinguishing direct from indirect miRNA-target interactions. We utilized CLIP procedure followed by screening with standard microarray platforms for miRNA and mRNA transcripts to demonstrate the miR-mRNA landscape and identify novel molecular targets for modulating T cell responses. Mixed cell culture was performed with T cells in the presence of syngeneic 57BL/6, or allogeneic BALB/c Dendritic cells or cultivated with anti-CD3e and anti-CD28 mAB for 24 h. Purified T cells were processd for CLIP procedures and followed RNA isolation (miReasy Kit and TRIzol LS) and hybridization on Exiqon 6th generation miRCURY LNA microRNA Arrays and Affymetrix microarrays. We sought to obtain that the specific changes in the expression of miRNAs and/or mRNAs from those responding to non-specific stimulation and in allogeneic activated T cells.