Modified epitope peptides targeting the porcine PD-1/PD-L1 interaction enhance cellular and humoral immune responses.

[IMPORTANCE] The programmed death-1 (PD-1)/programmed death ligand-1 (PD-L1) pathway transmits negative immunoregulatory signals. Blocking this pathway using peptides or antibodies can restore immunity.

[OBJECTIVE] To evaluate the immune function of epitope peptides interacting with porcine PD-1 or PD-L1.

[METHODS] We optimized and synthesized peptides (PD-L14QN-GF and PD-L14QN-AF) using the solid-phase method and assessed their effects on peripheral blood mononuclear cell (PBMC) proliferation and PD-1 and cytokine expression after porcine reproductive and respiratory syndrome virus (PRRSV) infection and on antibody responses to a porcine circovirus type 2 (PCV2) vaccine .

[RESULTS] The optimized peptides PD-L14QN-GF and PD-L14QN-AF exhibited lower binding free energy and higher stability when interacting with the PD-1 target protein. Under both PRRSV-infected and non-infected conditions , both peptides enhanced the proliferation of PBMCs, inhibited PRRSV RNA replication, and downregulated PD-1 transcription levels. Additionally, PD-L14QN-GF and PD-L14QN-AF upregulated the mRNA transcription and protein secretion of interleukin (IL)-2, IL-10, and interferon-γ to varying degrees. experiments demonstrated that PD-L14QN-GF significantly increased the antibody titer and seroconversion rate of the PCV2 vaccine.

[CONCLUSIONS AND RELEVANCE] PD-L14QN-GF and PD-L14QN-AF induced stronger immune responses than PD-L14. PD-L14QN-GF has potential as an immune-enhancing adjuvant.