RecombiMAb anti-mouse CCR8
(switched from Rat IgG2b, kappa)
Product Details
The C8Mab-2-CP080 monoclonal antibody is a recombinant, chimeric version of the original C8Mab-2 antibody. The variable domain sequences are identical but the constant region sequences have been switched from Rat IgG2b, Īŗ to Mouse IgG2a, Īŗ for use in murine models. Species-matched chimeric antibodies exhibit regulated effector functionsāincluding Fc receptor binding and complement activationāand result in less immunogenicity and formation of anti-drug antibodies (ADAs) than xenogenic antibodies in animal models. The anti-tumor activity of anti-CCR8 antibodies has been demonstrated to require Fc-mediated effector function with studies confirming the superior mouse IgG2a antibody binding to mouse FcĪ³RIII and mouse FcĪ³RIV is crucial for NK cell-mediated ADCC and macrophage-mediated ADCP in mice. The highly controlled sequence and lack of genetic drift in recombinant antibodies also provides more reliable and reproducible results over hybridoma derived antibodies.The C8Mab-2 monoclonal antibody recognizes the N-terminal region (1ā33 amino acids) of mouse C-C chemokine receptor type 8 (CCR8), also known as CKR-8, CDw198, CMKBRL2, CMKBR8, and GPRCY6. CCR8 is a seven-pass transmembrane chemokine receptor and a member of the G protein-coupled receptor (GPCR) family. CCR8 ligands include CCL1, CCL16, and CCL8 (mCCL8) or CCL18 (hCCL18, a functional analog of mouse CCL8). Human and mouse CCR8 as well as its primary ligand CCL1 are structurally related, and this ligand is critical for skin homing of T cells and the survival of the regulatory T cells (Tregs) as well as their chemotaxis into tumors. CCR8 is predominantly expressed on activated Tregs marking the most suppressive and proliferative Treg population residing in the TME. Regulatory T cells (Tregs) are immunosuppressive cells essential for maintaining peripheral immune tolerance and preventing harmful autoimmune responses. A deficiency in their number or function can lead to the development of autoimmune disorders. Conversely, an abundance of Tregs, particularly a high Treg-to-CD8+āT effector cell ratio, can hinder anti-tumor immune surveillance and promote cancer progression. CCR8, a surface receptor selectively expressed on activated Tregs within tumors, has emerged as a promising therapeutic target. Its selective expression offers the potential to enhance anticancer responses while minimizing the safety risks associated with earlier systemic Treg-targeting strategies. Recent in-vivo studies have documented the involvement of CCR8 in type 2 inflammatory diseases, including atopic dermatitis (AD) and allergic enteritis (AE). In the tumor microenvironment, CCR8+ Treg numbers directly correlate with an advanced state of cancer, and therapeutic depletion of CCR8+ tumor-infiltrating Tregs (ti-Tregs) is shown to exert antitumor immunity and synergism with anti-PD-1 therapy.
Specifications
| Isotype | Mouse IgG2a, Īŗ |
|---|---|
| Recommended Isotype Control(s) | RecombiMAb mouse IgG2a isotype control, unknown specificity |
| Recommended Dilution Buffer | InVivoPure pH 7.0 Dilution Buffer |
| Conjugation | This product is unconjugated. Conjugation is available via our Antibody Conjugation Services. |
| Reported Applications |
Flow Cytometry Western blot Immunofluorescence For information on in vivo applications, please contact (technicalservice@bioxcell.com) |
| Formulation |
PBS, pH 7.0 Contains no stabilizers or preservatives |
| Endotoxin |
<1EU/mg (<0.001EU/Ī¼g) Determined by LAL gel clotting assay |
| Aggregation |
<5% Determined by SEC |
| Purity |
>95% Determined by SDS-PAGE |
| Sterility | 0.2 Āµm filtration |
| Production | Purified from mammalian cell supernatant in an animal-free facility |
| Purification | Protein G |
| Molecular Weight | 150 kDa |
| Murine Pathogen Tests |
Ectromelia/Mousepox Virus: Negative Hantavirus: Negative K Virus: Negative Lactate Dehydrogenase-Elevating Virus: Negative Lymphocytic Choriomeningitis virus: Negative Mouse Adenovirus: Negative Mouse Cytomegalovirus: Negative Mouse Hepatitis Virus: Negative Mouse Minute Virus: Negative Mouse Norovirus: Negative Mouse Parvovirus: Negative Mouse Rotavirus: Negative Mycoplasma Pulmonis: Negative Pneumonia Virus of Mice: Negative Polyoma Virus: Negative Reovirus Screen: Negative Sendai Virus: Negative Theilerās Murine Encephalomyelitis: Negative |
| Storage | The antibody solution should be stored at the stock concentration at 4Ā°C. Do not freeze. |
Additional Formats
Recommended Products
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Recommended Isotype Control(s)
RecombiMAb mouse IgG2a isotype control, unknown specificity
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Recommended Dilution Buffer
InVivoPure pH 7.0 Dilution Buffer
Flow Cytometry
Kobayashi H, Suzuki H, Tanaka T, Kaneko MK, Kato Y. (2024). "Epitope Mapping of an Anti-Mouse CCR8 Monoclonal Antibody C8Mab-2 Using Flow Cytometry" Monoclon Antib Immunodiagn Immunother 43(4):101-107. PubMed
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 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, 1Ćalanine (or glycine) scanning and 2Ćalanine (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.
Roider HG, Hoff S, Tseng SY, Berndt S, Trautwein M, Filarsky K, Gritzan U, Camps J, Nadler WM, Grudzinska-Goebel J, Ellinger P, Pesch T, Soon CF, Geyer M, Gluske K, Stelte-Ludwig B, GorjƔnƔcz M. (2024). "Selective depletion of tumor-infiltrating regulatory T cells with BAY 3375968, a novel Fc-optimized anti-CCR8 antibody" Clin Exp Med 24(1):122. PubMed
Regulatory T cells (Tregs) are known to facilitate tumor progression by suppressing CD8+ T cells within the tumor microenvironment (TME), thereby also hampering the effectiveness of immune checkpoint inhibitors (ICIs). While systemic depletion of Tregs can enhance antitumor immunity, it also triggers undesirable autoimmune responses. Therefore, there is a need for therapeutic agents that selectively target Tregs within the TME without affecting systemic Tregs. In this study, as shown also by others, the chemokine (C-C motif) receptor 8 (CCR8) was found to be predominantly expressed on Tregs within the TME of both humans and mice, representing a unique target for selective depletion of tumor-residing Tregs. Based on this, we developed BAY 3375968, a novel anti-human CCR8 antibody, along with respective surrogate anti-mouse CCR8 antibodies, and demonstrated their in vitro mode-of-action through induction of potent antibody-dependent cellular cytotoxicity (ADCC) and phagocytosis (ADCP) activities. In vivo, anti-mouse CCR8 antibodies effectively depleted Tregs within the TME primarily via ADCP, leading to increased CD8+ T cell infiltration and subsequent tumor growth inhibition across various cancer models. This monotherapeutic efficacy was significantly enhanced in combination with ICIs. Collectively, these findings suggest that CCR8 targeting represents a promising strategy for Treg depletion in cancer therapies. BAY 3375968 is currently under investigation in a Phase I clinical trial (NCT05537740).
Flow Cytometry, Immunofluorescence
Tanaka T, Nanamiya R, Takei J, Nakamura T, Yanaka M, Hosono H, Sano M, Asano T, Kaneko MK, Kato Y. (2021). "Development of Anti-Mouse CC Chemokine Receptor 8 Monoclonal Antibodies for Flow Cytometry" Monoclon Antib Immunodiagn Immunother 40(2):65-70. PubMed
CC chemokine receptor 8 (CCR8) belongs to the class A of G protein-coupled receptor. It is highly expressed on Treg and T helper 2 (TH2) cells recruited to the inflammation site and is implicated in allergy and asthma. Recently, CCR8+Treg cells have been suggested to be a master regulator in the immunosuppressive tumor microenvironment; therefore, developing sensitive monoclonal antibodies (mAbs) for CCR8 has been desired. This study established a specific and sensitive mAb for mouse CCR8 (mCCR8), which is useful for flow cytometry by using the Cell-Based Immunization and Screening (CBIS) method. The established anti-mCCR8 mAb, C8Mab-2 (rat IgG2b, kappa), reacted with mCCR8-overexpressed Chinese hamster ovary-K1 (CHO/mCCR8) cells and P388 (mouse lymphoid neoplasma) or J774-1 (mouse macrophage-like) cells, which express endogenous mCCR8 by flow cytometry. C8Mab-2, which was established by the CBIS method, could be useful for elucidating the mCCR8-related biological response by flow cytometry.