Increased programmed death ligand (PD-L1) and cytotoxic T-lymphocyte antigen-4 (CTLA-4) expression is associated with metastasis and poor prognosis in malignant canine mammary gland tumours

Aberrant expression of immune check point molecules, programmed death ligand (PD-L1) and cytotoxic T-lymphocyte antigen-4 (CTLA-4) has been reported in many human cancers with increased protein and gene expression correlated with an aggressive behaviour in some neoplasms. Additionally, PD-L1 blockade has been shown to be an effective therapy for some human cancers. Canine mammary gland tumours have previously been shown to produce PD-L1 protein, but there are no previous studies investigating CTLA-4 in these common canine neoplasms. The present study investigated protein and gene expression of PD-L1 and CTLA-4 using immunohistochemistry and RT-PCR in 41 histologically-malignant, outcome-known CMGTs. The PD-L1 and CTLA-4 immunostaining scores of the mammary gland tumours that subsequently metastasised were significantly higher than those of tumours which did not metastasise (PD-L1: p = 0.005, CTLA-4: p = 0.003). Gene expression of PD-L1 and CTLA-4 was also significantly higher in tumours which subsequently metastasised (PD-L1: p = 0.023, CTLA-4: p = 0.022). Further, higher PD-L1 or CTLA-4 immunostaining scores correlated with shorter survival times of dogs (PD-L1: rs = - 0.42, p = 0.008, CTLA-4: rs = - 0.4, p = 0.01) while PD-L1 immunostaining was independently prognostic of survival time (Δ F = 4.9, p = 0.035). These findings suggest that higher protein and gene expression of PD-L1 and CTLA-4 by tumour cells increases the chances of metastasis and measuring these proteins may predict likely neoplasm behaviour. Additionally, if increased expression of these proteins promotes metastasis, blocking PD-L1 or CTLA-4 may be beneficial to treat canine mammary gland tumours.