C <sub>8</sub> Mab-2: An Anti-Mouse C–C Motif Chemokine Receptor 8 Monoclonal Antibody for Immunocytochemistry
Masaki Saito, Tomohiro Tanaka, Teizo Asano, Takuro Nakamura, Miyuki Yanaka, Saori Handa, Yu Komatsu, Yasuhiro Harigae, Nami Tateyama, Ren Nanamiya, Guanjie Li, Hiroyuki Suzuki, Mika K. Kaneko, Yukinari Kato
Abstract
C–C motif chemokine receptor 8 (CCR8) is a G protein-coupled receptor predominantly expressed in regulatory T (Treg) and T helper 2 cells. The evidence that CCR8 expression in Treg is increased in cancers, CCR8 increases migration activity of Treg, and CCR8 induces the anti-apoptotic activity in T cell leukemia and lymphoma suggests that CCR8 is associated with cancer development. Thus, developing a specific monoclonal antibody (mAb) for CCR8 is useful for diagnostic and therapeutic purposes and the anti-CCR8 mAb becomes a remarkable experimental tool for basic research. We previously developed an anti-mouse CCR8 (mCCR8) mAb called C 8 Mab-2 (rat IgG 2b , kappa) that was applicable to flow cytometric analysis for both endogenous and exogenous mCCR8. This study showed that C 8 Mab-2 and recombinant C 8 Mab-2 (recC 8 Mab-2) were specifically bound to exogenously expressed mCCR8 in mCCR8-overexpressed Chinese hamster ovary-K1 cells. In addition, we found that C 8 Mab-2 and recC 8 Mab-2 recognized endogenous mCCR8 in P388 (a mouse lymphocyte-like cell line) and J774-1 cells (a mouse macrophage-like cell line). These data demonstrate that C 8 Mab-2 and recC 8 Mab-2 are useful for immunocytochemical analysis.