Regulation of programmed death ligand 1 expression by interferon‐γ and tumour necrosis factor‐α in canine tumour cell lines

Abstract

AbstractExpression of programmed death ligand 1 (PD‐L1) on tumour cells provides an immune evasion mechanism by inducing suppression of cytotoxic T cells. Various regulatory mechanisms of PD‐L1 expression have been described in human tumours, however, little is known in canine tumours. To investigate whether inflammatory signalling is involved in PD‐L1 regulation in canine tumours, the effects of interferon (IFN)‐γ and tumour necrosis factor (TNF)‐α treatment were examined in canine malignant melanoma cell lines (CMeC and LMeC) and an osteosarcoma cell line (HMPOS). The protein level of PD‐L1 expression was upregulated by IFN‐γ and TNF‐α stimulation. Upon IFN‐γ stimulation, all cell lines showed an increase in expression of PD‐L1, signal transducer and activator of transcription (STAT)1, STAT3 and genes regulated by STAT activation. Upregulated expression of these genes was suppressed by the addition of a JAK inhibitor, oclacitinib. Contrastingly, upon TNF‐α stimulation, all cell lines exhibited higher gene expression of the nuclear factor kappa B (NF‐κB) gene RELA and genes regulated by NF‐κB activation, whereas expression of PD‐L1 was upregulated in LMeC only. Upregulated expression of these genes was suppressed by the addition of an NF‐κB inhibitor, BAY 11‐7082. The expression level of cell surface PD‐L1 induced by IFN‐γ and TNF‐α treatment was reduced by oclacitinib and BAY 11‐7082, respectively, indicating that upregulation of PD‐L1 expression by IFN‐γ and TNF‐α stimulation is regulated via the JAK‐STAT and NF‐κB signalling pathways, respectively. These results provide insights into the role of inflammatory signalling in PD‐L1 regulation in canine tumours.