Differential activation of OT-1 CD8+ T cells by CD40-activated B cells and dendritic cells

To determine if CD40-activated B or dendritic cells can induce a similar or different genetic profile in OT-1 CD8+ T cells in early time points following activation CD40-activated B and dendritic cells were cultured in vitro, matured with LPS and loaded with the SIINFEKL peptide (OVA 257-264). OT-1 T cells (CD8+ specifically recognizing the SIINFEKL peptide in the context of H2-Kb) were cocultured either with peptide pulsed CD40-activated B cells or dendritic cells for 24h. Following coculture, activated OT-1 T cells were sorted based on high expression of CD69 (CD8+CD69+). RNA from sorted cells was extracted using TRIZOL followed Qiagen RNA Cleanup and processed for hybridization to the Illumina mouse-ref8-v2.0 expression beadchip according to the manufacturers protocol.