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Development of a Novel Anti-Mouse CCR4 Monoclonal Antibody (CMab-1) by N-Terminal Peptide Immunization.

The CC chemokine receptor type-4 (CCR4) belongs to the G-protein-coupled receptor superfamily, expressed on the cell surface of T cells and its malignancy. Two CCR4 ligands (CCL17 and CCL22) bind to CCR4 that mediate physiological and pathological functions of T cell immune responses. Anti-CCR4 monoclonal antibody (mAb) mogamulizumab is approved for adult T cell leukemia/lymphoma and cutaneous T cell lymphomas. In addition, mogamulizumab can deplete regulatory T cells, implying the application to solid tumors as an immunomodulator. Therefore, the development of sensitive mAbs for CCR4 has been desired for basic research, diagnosis, and therapy. In this study, a specific, and sensitive anti-mouse CCR4 (mCCR4) mAb, CMab-1 (rat IgG, kappa), was established using N-terminal peptide immunization. CMab-1 reacted with mCCR4-overexpressed Chinese hamster ovary (CHO)-K1 cells, P388 (mouse lymphoid neoplasm), and J774-1 (mouse macrophage-like) cells in flow cytometry. Kinetic analyses using flow cytometry showed that s of CMab-1 for CHO/mCCR4, P388, and J774-1 cells were 4.2 × 10 M, 5.4 × 10 M, and 1.1 × 10 M, respectively. CMab-1 could be a valuable tool for elucidating mCCR4-related biological responses.
Junko Takei, Hiroyuki Suzuki, Teizo Asano, Tomohiro Tanaka, Mika K Kaneko, Yukinari Kato

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