Kobayashi Hiyori, Asano Teizo, Suzuki Hiroyuki, Tanaka Tomohiro, Yoshikawa Takeo, Kaneko Mika K, Kato Yukinari
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. 2023 Feb;42(1):15-21. doi: 10.1089/mab.2022.0032. Epub 2022 Dec 13.
The CC chemokine receptor 9 (CCR9), also known as CD199, is one of chemokine receptors. The CC chemokine ligand 25 (CCL25) is known to be the only ligand for CCR9. The CCR9-CCL25 interaction plays important roles in chemotaxis of lymphocytes and tumor cell migration. Therefore, CCR9-CCL25 axis is a promising target for tumor therapy and diagnosis. In this study, we established a sensitive and specific monoclonal antibody (mAb) against mouse CCR9 (mCCR9) using N-terminal peptide immunization method. The established anti-mCCR9 mAb, CMab-24 (rat immunoglobulin [IgG], kappa), reacted with mCCR9-overexpressed Chinese hamster ovary-K1 (CHO/mCCR9) and mCCR9-endogenously expressed cell line, RL2, through flow cytometry. Kinetic analyses using flow cytometry showed that the dissociation constants () of CMab-24 for CHO/mCCR9 and RL2 cell lines were 6.0 × 10 M and 4.7 × 10 M, respectively. Results indicated that CMab-24 is useful for detecting mCCR9 through flow cytometry, thereby providing a possibility for targeting mCCR9-expressing cells experiments.
CC趋化因子受体9（CCR9），也称为CD199，是趋化因子受体之一。已知CC趋化因子配体25（CCL25）是CCR9的唯一配体。CCR9-CCL25相互作用在淋巴细胞的趋化作用和肿瘤细胞迁移中起重要作用。因此，CCR9-CCL25轴是肿瘤治疗和诊断的一个有前景的靶点。在本研究中，我们使用N端肽免疫法建立了一种针对小鼠CCR9（mCCR9）的灵敏且特异的单克隆抗体（mAb）。所建立的抗mCCR9单克隆抗体CMab-24（大鼠免疫球蛋白[IgG]，κ），通过流式细胞术与过表达mCCR9的中国仓鼠卵巢-K1（CHO/mCCR9）细胞和内源性表达mCCR9的细胞系RL2发生反应。使用流式细胞术进行的动力学分析表明，CMab-24对CHO/mCCR9和RL2细胞系的解离常数（）分别为6.0×10 M和4.7×10 M。结果表明，CMab-24可用于通过流式细胞术检测mCCR9，从而为靶向表达mCCR9的细胞实验提供了可能性。
