Development of a Novel Anti-Mouse CCR6 Monoclonal Antibody (C 6 Mab-13) by N-Terminal Peptide Immunization.

The CC chemokine receptor 6 (CCR6) is a G protein-coupled receptor family member that is highly expressed in B lymphocytes, certain subsets of effector and memory T cells, and immature dendritic cells. CCR6 has only one chemokine ligand, CCL20. The CCL20-CCR6 axis has been recognized as a therapeutic target for autoimmune diseases and tumor. This study developed specific monoclonal antibodies (mAbs) against mouse CCR6 (mCCR6) using the peptide immunization method. The established anti-mCCR6 mAb, C 6 Mab-13 (rat IgG 1 , kappa), reacted with mCCR6-overexpressed Chinese hamster ovary-K1 (CHO/mCCR6), and mCCR6-endogenously expressed P388 (mouse lymphoid neoplasma) and J774-1 (mouse macrophage-like) cells in flow cytometry. The dissociation constant ( K D ) of C 6 Mab-13 for CHO/mCCR6 cells was determined to be 2.8 × 10 -9 M, indicating that C 6 Mab-13 binds to mCCR6 with high affinity. In summary, C 6 Mab-13 is useful for detecting mCCR6-expressing cells through flow cytometry.