Department of Antibody Drug Development, Tohoku University Graduate School of Medicine, Sendai, Japan.
Monoclon Antib Immunodiagn Immunother. 2023 Aug;42(4):117-124. doi: 10.1089/mab.2023.0004. Epub 2023 Jul 10.
CC chemokine receptor 6 (CCR6) is a member of the G-protein-coupled receptor family that is highly expressed in B lymphocytes, effector and memory T cells, regulatory T cells, and immature dendritic cells. CCR6 has been revealed to have important functions in many pathological conditions, such as cancer, intestinal bowel disease, psoriasis, and autoimmune diseases. The only CCR6 chemokine ligand, CC motif chemokine ligand 20 (CCL20), is also involved in pathogenesis by interacting with CCR6. The CCL20/CCR6 axis is drawing attention as an attractive therapeutic target for various diseases. In this study, we developed novel monoclonal antibodies (mAbs) against human CCR6 (hCCR6) using the peptide immunization method, which are applicable to flow cytometry and immunohistochemistry. The established anti-hCCR6 mAb, clone CMab-19 (mouse IgG, kappa), reacted with hCCR6-overexpressed Chinese hamster ovary-K1 (CHO/hCCR6), human liver carcinoma (HepG2), and human differentiated hepatoma (HuH-7) cells in flow cytometry. The dissociation constant () of CMab-19 was determined as 3.0 × 10 M for CHO/hCCR6, 6.9 × 10 M for HepG2, and 1.8 × 10 M for HuH-7. Thus, CMab-19 could bind to exogenously and endogenously expressed hCCR6 with extremely high affinity. Furthermore, CMab-19 could stain formalin-fixed paraffin-embedded lymph node tissues from a patient with non-Hodgkin lymphoma by immunohistochemistry. Therefore, CMab-19 is suitable for detecting hCCR6-expressing cells and tissues and could be useful for pathological analysis and diagnosis.
CC 趋化因子受体 6（CCR6）是 G 蛋白偶联受体家族的成员，在 B 淋巴细胞、效应和记忆 T 细胞、调节性 T 细胞和未成熟树突状细胞中高度表达。CCR6 在许多病理条件下具有重要功能，如癌症、肠道疾病、银屑病和自身免疫性疾病。唯一的 CCR6 趋化因子配体，CC 基序趋化因子配体 20（CCL20），通过与 CCR6 相互作用也参与发病机制。CCL20/CCR6 轴作为各种疾病有吸引力的治疗靶点引起了关注。在这项研究中，我们使用肽免疫法开发了针对人 CCR6（hCCR6）的新型单克隆抗体（mAb），可用于流式细胞术和免疫组织化学。建立的抗 hCCR6 mAb，克隆 CMab-19（鼠 IgG，kappa），在流式细胞术检测中与人 CCR6 过表达的中国仓鼠卵巢-K1（CHO/hCCR6）、人肝癌（HepG2）和人分化肝癌（HuH-7）细胞反应。CMab-19 的解离常数（）确定为 CHO/hCCR6 为 3.0×10 M，HepG2 为 6.9×10 M，HuH-7 为 1.8×10 M。因此，CMab-19 可以与外源性和内源性表达的 hCCR6 以极高的亲和力结合。此外，CMab-19 可以通过免疫组织化学染色非霍奇金淋巴瘤患者的福尔马林固定石蜡包埋淋巴结组织。因此，CMab-19 适用于检测表达 hCCR6 的细胞和组织，可用于病理分析和诊断。
