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

Molecular networks based on genome-wide mRNA and miRNA expression in activated T cells are interated 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 mAB9 for 24 h. Purified T cells were processde for CLIP procedures and followed by RNA isolation (TRIzol LS and miReasy Kit) and hybridization on Affymetrix microarrays and Exiqon 6th generation miRCURY LNA microRNA Arrays . We sought to obtain that the specific changes in the expression of miRNAs and/or mRNAs in allogeneic activated T cells during hematopoietic cells transplantation and that would be distinct from those responding to non-specific stimulation.