Anti-Mouse LPAM-1- Purified in vivo PLATINUM™ Functional Grade

Common In Vivo Applications of Clone DATK32 in Mice

Scientific Background
Clone DATK32 is a rat IgG2a monoclonal antibody that specifically targets mouse LPAM-1, also known as integrin α4β7, a receptor expressed on peripheral lymphocytes, thymocyte subsets, and bone marrow progenitors. This integrin is critical for lymphocyte adhesion and migration to mucosal sites, especially the intestine and gut-associated lymphoid tissues, by recognizing ligands such as VCAM-1, MAdCAM-1, and fibronectin.

Key In Vivo Applications

• Blockade of Lymphocyte Homing: The most common and clinically relevant in vivo application of DATK32 is blocking the adhesion of lymphocytes to endothelial cells expressing VCAM-1, MAdCAM-1, or fibronectin—thereby inhibiting lymphocyte migration to mucosal tissues. This has direct implications for studies of inflammatory bowel disease, gut immunity, and mucosal homing responses.
• Inhibition of Mucosal Immunity: By disrupting the integrin α4β7–MAdCAM-1 interaction, DATK32 is used to impair the gut-tropic homing of lymphocytes, reducing inflammatory responses and lymphocyte recruitment in mouse models of colitis, experimental autoimmune encephalomyelitis (EAE), and other mucosal immune disorders.
• Functional Analog of Clinical Therapeutics: DATK32 is often used preclinically as a mouse analog of vedolizumab (a humanized anti-α4β7 antibody), which is approved for treating inflammatory bowel diseases in humans.
• Studies of Lymphocyte Trafficking: DATK32 has been used to elucidate the mechanisms of lymphocyte recruitment to mucosal sites, including Peyer’s patches and intestinal lamina propria, by blocking adhesion in vivo and enabling researchers to assess the role of α4β7 integrin in leukocyte extravasation and inflammatory responses.
• Combination with Lineage-Specific Markers: In experimental settings, DATK32 has been used alongside lineage-specific antibodies (e.g., CD45R) and isotype controls to dissect the roles of specific lymphocyte subsets in mucosal immunity and inflammation.

Experimental Considerations

• Dose and Specificity: DATK32 is highly specific for mouse LPAM-1 and has been validated for use in both blocking (neutralizing) and detection (flow cytometry, immunohistochemistry) applications in vivo.
• Endotoxin Grade: For in vivo studies, low-endotoxin, functional-grade formulations are essential to avoid confounding immune responses.
• Isotype Controls: Rat IgG2a isotype controls are recommended to distinguish specific effects from non-specific antibody-mediated effects.

Summary Table

Conclusion

In vivo, clone DATK32 is primarily used to block the adhesion and trafficking of lymphocytes to mucosal tissues in mice, serving as a critical tool for studying gut immunity, inflammatory diseases, and the mechanisms of leukocyte homing. Its applications mirror those of clinical anti-α4β7 therapies, making it a valuable reagent in preclinical immunology research.

Other commonly used antibodies and proteins in studies featuring DATK32 (anti-mouse integrin α4β7/LPAM-1) include:

• Natural ligands for α4β7 (LPAM-1):

  • VCAM-1 (CD106)
  • MAdCAM-1
  • Fibronectin
    These proteins are frequently used for cell adhesion assays and functional blocking studies with DATK32.

• Lineage or subset marker antibodies:

  • CD45R/B220 (B cell marker)
  • CD3 (T cell marker)
  • CD4/CD8 (T cell subset markers)
  • CD103 (Integrin αE) (Often used for defining gut-resident lymphocyte populations, as β7 also pairs with αE to form αEβ7).

• Isotype controls:

  • Rat IgG2a, κ
    These are used for controlling non-specific binding in flow cytometry and immunohistochemistry.

• Therapeutic or blocking antibodies and analogs:

  • Vedolizumab (Anti-human integrin α4β7, used as a comparative agent in mouse studies; humanized analog for clinical relevance).
  • Etrolizumab (Anti-human integrin β7, often cited in comparative studies).

• Other integrin antibodies:

  • Anti-integrin β7 (distinguishes between β7-containing dimers such as α4β7 and αEβ7).
  • FIB504, FIB27, rhuMAb Beta7 (Additional monoclonal antibodies targeting β7).

These reagents are commonly used in combination with DATK32 for:

• Cell subset and phenotyping panels in flow cytometry
• Adhesion and migration assays employing ligand-coated surfaces
• Functional blocking experiments
• Immunohistochemistry and tissue staining
• Comparative studies with therapeutic monoclonal antibodies

For precise experimental design, other inclusion markers may depend on the tissue or cell type being studied and on whether the experiment is in vitro or in vivo.

Clone DATK32 is a rat monoclonal antibody specific for mouse integrin α4β7 (LPAM-1), widely used in immunology research to study and block lymphocyte adhesion, trafficking, and mucosal immune responses.

Key findings from scientific literature and product documentation using clone DATK32 include:

• Functional Blocking of Integrin α4β7: DATK32 effectively binds to and blocks the function of α4β7 integrin, inhibiting lymphocyte adhesion to its ligands—VCAM-1, MAdCAM-1, and fibronectin—both in vitro and in vivo.

• Applications in Flow Cytometry and Neutralization: DATK32 is validated for use in flow cytometry (for staining and quantifying α4β7+ cells), blocking/neutralization, and immunoprecipitation. It can induce aggregation of T cell lymphoma lines and block LPAM-1-mediated cell adhesion.

• Role in Lymphocyte Homing: LPAM-1 (α4β7) is crucial for lymphocyte trafficking to mucosal tissues. DATK32 helps elucidate the mechanism by which lymphocytes home to gut-associated lymphoid tissues, including Peyer’s patches, via interaction with MAdCAM-1.

• Preclinical Use in Inflammatory Bowel Disease (IBD) Models:

  • Targeted Delivery Increases Local Tissue Concentration: Intracolic (IC) administration of DATK32 leads to higher drug levels in colon tissue and contents, but lower plasma exposure compared to intraperitoneal (IP) delivery.
  • Reduction of Pathogenic Memory T cells: Targeted IC DATK32 significantly reduced α4β7 memory T cells in Peyer’s patches and mesenteric lymph nodes during experimental colitis.
  • Proof-of-Concept for Local Therapy: Findings support the idea that targeted, topical antibody delivery may improve pharmacodynamic effects and therapeutic efficacy in IBD by maximizing drug concentration at inflamed mucosal sites and minimizing systemic exposure.

• Mechanistic Studies in Mucosal Immunity and Tumor Biology: DATK32 has been used to demonstrate the importance of the α4β7 integrin:

  • In the transmigration of leukocytes during inflammation,
  • In the modeling of mucosal immune responses and autoimmunity,
  • And in studies on lymphocyte interaction with endothelium in tumor biology.

• Therapeutic Antibody Development: The mechanism targeted by DATK32 (α4β7 blockade) has translational relevance—other antibodies targeting the same pathway, such as vedolizumab, are used clinically to treat IBD.

In summary, DATK32 is a widely cited and validated research tool for studying lymphocyte migration, mucosal immunity, and antibody-based therapies—especially in murine models of inflammation and autoimmunity.

Dosing regimens of clone DATK32—an anti-mouse α4β7 integrin monoclonal antibody—vary across mouse models by dose, frequency, and route, most commonly depending on experimental aims and disease context.

Common dosing regimens:

• Acute DSS-Colitis model (BALB/c mice):

  • Intraperitoneal (IP): 25 mg/kg every 3 days (Q3D).
  • Intracecal (IC): 25 mg/kg either every 3 days (Q3D) or daily (QD); lower dose of 5 mg/kg IC daily (QD) also tested.
  • All treatments were administered from day 0 to day 14, with final analysis on day 14.
  • Drug levels remained elevated in colon tissue and content for up to 48 hours following dosing, with significant concentrations sustained for 8–24 hours, depending on route and dose.

• Experimental Autoimmune Encephalomyelitis (EAE) model (C57BL/6 mice):

  • Intraperitoneal (IP): 10 mg/kg, administered twice weekly.
  • This regimen was designed for depletion of CD4+ T cells in EAE-induced mice.

Key variables that differ by model:

• Dose: 25 mg/kg (IP/IC) used in colitis; 10 mg/kg (IP) in EAE.
• Frequency: Every 3 days or daily (colitis); twice weekly (EAE).
• Route: Both IP and IC (colitis); IP only (EAE).
• Objective: Immunomodulation in colitis versus T cell depletion in EAE.

Summary Table: DATK32 Dosing Regimens in Mouse Models

Noteworthy study details:

• Route and frequency impact colon drug levels and pharmacodynamics—intracecal (IC) administration led to higher concentrations locally compared to systemic (IP) dosing, with the highest tissue levels seen in proximal colon after IC delivery.
• Lower IC daily doses (5 mg/kg) are sometimes explored for dose-response or toxicity assessment.

Conclusion:
DATK32 regimens in mice are tailored to disease models and experimental goals, with IP and IC routes, doses from 5–25 mg/kg, and frequencies from daily to twice weekly. Individual studies may adjust dosing based on strain, disease activity, and desired immunological outcomes, so optimal dosing must be empirically determined for each context.