Kobayashi, H.; Suzuki, H.; Tanaka, T.; Kaneko, M.K.; Kato, Y. Epitope Mapping of an Anti-mouse CCR8 Monoclonal Antibody C8Mab-2 Using Flow Cytometry. Preprints2024, 2024011895. https://doi.org/10.20944/preprints202401.1895.v1
APA Style
Kobayashi, H., Suzuki, H., Tanaka, T., Kaneko, M.K., & Kato, Y. (2024). Epitope Mapping of an Anti-mouse CCR8 Monoclonal Antibody C<sub>8</sub>Mab-2 Using Flow Cytometry. Preprints. https://doi.org/10.20944/preprints202401.1895.v1
Chicago/Turabian Style
Kobayashi, H., Mika K. Kaneko and Yukinari Kato. 2024 "Epitope Mapping of an Anti-mouse CCR8 Monoclonal Antibody C<sub>8</sub>Mab-2 Using Flow Cytometry" Preprints. https://doi.org/10.20944/preprints202401.1895.v1
Abstract
The C-C motif chemokine receptor 8 (CCR8) is highly and selectively expressed in regulatory T (Treg) cells and is associated with tumor progression. The massive accumulation of Treg cells into tumors suppresses the effector function of CD8+ cells against tumor cells. Therefore, selective depletion of Treg cells using anti-CCR8 monoclonal antibodies (mAbs) reinvigorates antitumor immune responses and improves responses to cancer immunotherapy. Previously, we developed an anti-mouse CCR8 (mCCR8) mAb, C8Mab-2, using the Cell-Based Immunization and Screening (CBIS) method. In this study, the binding epitope of C8Mab-2 was investigated using flow cytometry. The mCCR8 extracellular domain-substituted mutant analysis showed that C8Mab-2 recognizes the N-terminal region (1–33 amino acids) of mCCR8. Next, 1×alanine (or glycine) scanning and 2×alanine (or glycine) scanning were conducted in the N-terminal region. The results revealed that the 17-DFFTAP-22 sequence is important for the recognition by C8Mab-2, and Thr20 is a central amino acid of the epitope.These results revealed the involvement of the N-terminus of mCCR8 in the recognition by C8Mab-2.
Medicine and Pharmacology, Oncology and Oncogenics
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