Development of canine PD-1/PD-L1 specific monoclonal antibodies and amplification of canine T cell function
Development of canine PD-1/PD-L1 specific monoclonal antibodies and amplification of canine T cell function
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DOI：
10.1371/journal.pone.0235518
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发表时间：
2020-07-02
期刊：
影响因子：
3.7
通讯作者：
McSorley, Stephen J.
中科院分区：
文献类型：
--
作者：
Choi, Jin Wook;Withers, Sita S.;McSorley, Stephen J.
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既定且有效的治疗策略，并为兽医肿瘤学带来了希望。我们报告了犬 PD-1 和 PD-L1 特异性单克隆抗体的生成和表征。最初评估抗体阻断重组犬 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名。选定的抗体能够通过流式细胞术和蛋白质印迹检测犬组织上的 PD-1 和 PD-L1。抗 PD-L1 适用于福尔马林固定石蜡包埋的犬组织的免疫细胞化学，抗 PD-1 适用于免疫组织化学，表明该抗体可用于存档的组织。此外，抗 PD-L1 (JC071) 显示，在用肽聚糖或脂多糖刺激后，犬单核细胞上的 PD-L1 表达显着增加。这些抗体一起显示出对天然犬配体的特异性，可用于各种潜在的诊断应用。重要的是，多种 PD-L1 特异性抗体在犬外周血单核细胞 (PBMC) 刀豆蛋白 A (Con A) 刺激试验中放大了 IFN-γ 的产生，证明了其功能活性。
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-gamma production in a canine peripheral blood mononuclear cells (PBMC) concanavlin A (Con A) stimulation assay, demonstrating functional activity.