Expression of FOXJ1 and ITGB4 is Activated upon KLH and LTA Stimulation in the DT40 Cell Line

The immune response is regulated by multiple genes. In chickens, a panel of genes associated with humoral immune responses against Keyhole limpet hemocyanin (KLH), lipopolysaccharide (LPS) and lipoteichoic acid (LTA) was detected on chromosomes 9, 14, 18 and Z. In this study, we analyzed the transcriptional activity of these genes in the DT40 cell line stimulated with KLH, LPS and LTA antigens in vitro. mRNA synthesis of the genes of interest was analyzed at 3, 6, 9, and 24 hours post-treatment, using reverse transcription quantitative PCR (RT-qPCR). The results indicate that KLH and LTA antigens were the strongest stimuli of gene expression, whereas LPS treatment did not activate mRNA synthesis of any of the studied genes. Relative analysis of gene expression showed that KLH and LTA treatment triggered up-regulation of forkhead box J1 (FOXJ1) and integrin beta 4 (ITGB4) genes at 24 hours post-treatment (P<0.05). Western blot analysis confirmed expression of FOXJ1 and ITGB4 genes by the DT40 cell line also at the protein level. Finally, the functional relations between LTA stimulation and FOXJ1 and ITGB4 genes suggests activation of the TLR signaling pathway, but also involvement of scavenger receptors in LTA recognition. This approach provided additional information on transcriptional activity of the genes selected in the genetic association studies.