Figure 1

Analysis of immunoinhibitory functions by chicken PD-1/PD-L pathway.(A) The expression of PD-1 mRNA was determined in MSB1, HP1, Lee1 cell, and PBMCs from uninfected chickens by real-time RT-PCR. The degree of the PD-1 mRNA expression was expressed relative to the mean basal value in PBMCs after normalization to that of β-actin mRNA. (B) The expression levels of IFN-γ mRNA in Lee1 cells co-cultured with DF-1 cells were determined by real-time RT-PCR. DF-1 cells were transfected with pCMV-Tag1 or pCMV-PDL1. The degree of the PD-1 and IFN-γ mRNA expression was expressed relative to the mean basal value in the presence of pCMV-Tag1 after normalization to that of β-actin mRNA. Three independent experiments were performed in triplicate. Error bars indicate standard deviations. The significant differences were determined by Student’s paired t-test between types of expression plasmids (*P < 0.05). (C) Transient expression of PD-L1 construct in DF-1 cells was determined by western blotting. Blots were also probed with an anti-actin monoclonal antibody as a loading control.