Defining the molecular basis of dendritic cell mediated tolerance. Mus musculus

Using a TCR transgenic transplantation model a series of differently modulated bmDC (IL10, TGFb and VD3 treated, +/-LPS) were established that could mediate tolerance following adaptive transfer. These modified cells, in contrast to untreated 'immature' bmDC retained their capacity to tolerise following exposure to LPS. The unmodified, non-tolerogenic, LPS-treated cells provided a comparator population to facilitate investigation into molecular expression patterns that correlate with functional phenotype. Keywords: pattern searching, immune tolerance, dendritic cell Overall design: To investigate transcriptional-based explainations for the shared tolerancing-inducing phenotype of bmDC cultured under different growth contexts, pattern searching was performed across 29 SAGE libraries, using bmDC+LPS (one long and one short SAGE library) as a non-tolerant comparator (GSM580, GSM767, GSM1681-2, GSM3677-79, GSM3681-6, GSM3824-29, GSM3832-7, GSM132450 and GSM160043, two independant LPS+ comparator libraries were included while all the other libraries were single biological/technical replicates) . Pattern searching of the SAGE data was combined with equivalent analysis of single channel micro-array data (E-MEXP-845) to select genes to be included on Taqman low density microfluidic card. QPCR was used to verify expression patterns using three biological replicates of the complete cohort of tolerogenic and the non-tolerogenic comparator DC populations.