Center for Immunology and Infectious Diseases, Department of Anatomy, Physiology and Cell Biology, School of Veterinary Medicine, University of California, Davis, California, United States of America.
Center for Companion Animal Health, Department of Surgical and Radiological Sciences, School of Veterinary Medicine, University of California, Davis, California, United States of America.
PLoS One. 2020 Jul 2;15(7):e0235518. doi: 10.1371/journal.pone.0235518. eCollection 2020.
Interruption of the programmed death 1 (PD-1) / programmed death ligand 1 (PD-L1) pathway is an established and effective therapeutic strategy in human oncology and holds promise for veterinary oncology. We report the generation and characterization of monoclonal antibodies specific for canine PD-1 and PD-L1. Antibodies were initially assessed for their capacity to block the binding of recombinant canine PD-1 to recombinant canine PD-L1 and then ranked based on efficiency of binding as judged by flow cytometry. Selected antibodies were capable of detecting PD-1 and PD-L1 on canine tissues by flow cytometry and Western blot. Anti-PD-L1 worked for immunocytochemistry and anti-PD-1 worked for immunohistochemistry on formalin-fixed paraffin embedded canine tissues, suggesting the usage of this antibody with archived tissues. Additionally, anti-PD-L1 (JC071) revealed significantly increased PD-L1 expression on canine monocytes after stimulation with peptidoglycan or lipopolysaccharide. Together, these antibodies display specificity for the natural canine ligand using a variety of potential diagnostic applications. Importantly, multiple PD-L1-specific antibodies amplified IFN-γ production in a canine peripheral blood mononuclear cells (PBMC) concanavlin A (Con A) stimulation assay, demonstrating functional activity.
阻断程序性死亡 1（PD-1）/程序性死亡配体 1（PD-L1）通路是人类肿瘤学中一种已确立且有效的治疗策略，并有望应用于兽医肿瘤学。我们报告了用于检测犬 PD-1 和 PD-L1 的单克隆抗体的产生和特性。首先评估了抗体阻断重组犬 PD-1 与重组犬 PD-L1 结合的能力，然后根据流式细胞术判断的结合效率进行排序。选择的抗体能够通过流式细胞术和 Western blot 检测犬组织中的 PD-1 和 PD-L1。免疫细胞化学使用抗 PD-L1（JC071），福尔马林固定石蜡包埋的犬组织的免疫组织化学使用抗 PD-1，这表明该抗体可用于存档组织。此外，抗 PD-L1（JC071）在肽聚糖或脂多糖刺激后，犬单核细胞上的 PD-L1 表达显著增加。总之，这些抗体使用多种潜在的诊断应用显示出对天然犬配体的特异性。重要的是，在犬外周血单核细胞（PBMC）刀豆蛋白 A（Con A）刺激试验中，多种 PD-L1 特异性抗体可扩增 IFN-γ 的产生，证明了其功能活性。
