Engineering a High-Affinity PD-1 Peptide for Optimized Immune Cell-Mediated Tumor Therapy

Abstract

The purpose of this study was to optimize a peptide (nABP284) that binds to PD-1 by a computer-based protocol in order to increase its affinity. Then, this study aimed to determine the inhibitory effects of this peptide on cancer immune escape by coculturing improving cytokine-induced killer (ICIK) cells with cancer cells. Materials and MethodsnABP284 that binds to PD-1 was identified by phage display technology in our previous study.AutoDock and PyMOL were used to optimize the sequence of nABP284 to design a new peptide (nABPD1). Immunofluorescence was used to demonstrate that the peptides bound to PD-1. Surface plasmon resonance (SPR) was used to measure the binding affinity of the peptides. The blocking effect of the peptides on PD-1 was evaluated by a neutralization experiment with human recombinant PD-L1 protein. The inhibition of activated lymphocytes by cancer cells was simulated by coculturing of human acute T lymphocytic leukemia cells (Jurkat T cells) with human tongue squamous cell carcinoma cells (Cal27 cells). The anticancer activities were determined by coculturing ICIK cells with Cal27 cells in vitro. ResultsA high-affinity peptide (nABPD1, KD=11.9 nM) for PD-1 was obtained by optimizing the nABP284 peptide (KD=11.8 µM). nABPD1 showed better efficacy than nABP284 in terms of increasing the secretion of IL-2 by Jurkat T cells and enhancing the in vitro antitumor activity of ICIK cells. ConclusionnABPD1 possesses higher affinity for PD-1 than nABP284, which significantly enhances its ability to block the PD-1/PD-L1 interaction and to increase ICIK cell-mediated antitumor activity by armoring ICIK cells.

Progress of research on PD-1/PD-L1 in leukemia

“…A new peptide, nABPD1, is designed to specifically bind PD-1. It enhances cytokine-induced killer (ICIK) cell-mediated antitumor activity by protecting ICIK cells through blockade of PD-1 signaling ( 44 ). There is a lack of clinical studies on the use of PD-1/PD-L1 mAbs in ALL, especially in young people, and as far as the current studies are concerned, they show unsatisfactory results for MRD in adults(median age is 52.5y) with ALL ( 132 ).…”