Department of Molecular Pharmacology, Tohoku University Graduate School of Medicine, Sendai, Japan.
Department of Antibody Drug Development, Tohoku University Graduate School of Medicine, Sendai, Japan.
Monoclon Antib Immunodiagn Immunother. 2022 Apr;41(2):101-109. doi: 10.1089/mab.2021.0067.
Ferrets () have been used as small animal models to investigate severe acute respiratory syndrome coronaviruses (SARS-CoV and SARS-CoV-2) infections. Pathological analyses of these tissue samples, including those of the lung, are, therefore, essential to understand the pathogenesis of SARS-CoVs and evaluate the action of therapeutic monoclonal antibodies (mAbs) against this disease. However, mAbs that recognize ferret-derived proteins and distinguish between specific cell types, such as lung epithelial cells, are limited. Podoplanin (PDPN) has been identified as an essential marker in lung type I alveolar epithelial cells, kidney podocytes, and lymphatic endothelial cells. In this study, an anti-ferret PDPN (ferPDPN) mAb PMab-292 (mouse IgG, kappa) was established using the Cell-Based Immunization and Screening (CBIS) method. PMab-292 recognized ferPDPN-overexpressed Chinese hamster ovary-K1 (CHO/ferPDPN) cells by flow cytometry and Western blotting. The kinetic analysis using flow cytometry showed that the of PMab-292 for CHO/ferPDPN was 3.4 × 10 M. Furthermore, PMab-292 detected lung type I alveolar epithelial cells, lymphatic endothelial cells, and glomerular/Bowman's capsule in the kidney using immunohistochemistry. Hence, these results propose the usefulness of PMab-292 in analyzing ferret-derived tissues for SARS-CoV-2 research.
雪貂（ferrets）已被用作研究严重急性呼吸系统综合征冠状病毒（SARS-CoV 和 SARS-CoV-2）感染的小动物模型。因此，对这些组织样本（包括肺组织样本）进行病理学分析对于了解 SARS-CoV 的发病机制以及评估针对这种疾病的治疗性单克隆抗体（mAbs）的作用至关重要。然而，能够识别雪貂来源的蛋白并区分特定细胞类型（如肺上皮细胞）的 mAbs 十分有限。Podoplanin（PDPN）已被鉴定为肺 I 型肺泡上皮细胞、肾脏足细胞和淋巴管内皮细胞中的重要标志物。在这项研究中，使用基于细胞的免疫接种和筛选（CBIS）方法建立了针对雪貂 PDPN（ferPDPN）的 mAb PMab-292（鼠 IgG，kappa）。PMab-292 通过流式细胞术和 Western blot 识别过表达 ferPDPN 的中国仓鼠卵巢-K1（CHO/ferPDPN）细胞。通过流式细胞术进行的动力学分析表明，PMab-292 与 CHO/ferPDPN 的亲和力为 3.4×10-8M。此外，PMab-292 通过免疫组织化学检测到肺 I 型肺泡上皮细胞、淋巴管内皮细胞和肾脏中的肾小球/鲍曼氏囊。因此，这些结果表明 PMab-292 可用于分析 SARS-CoV-2 研究中的雪貂来源组织。
