Anti-Mouse TIM-1 (CD365) [Clone RMT1-10] — Purified in vivo PLATINUM™ Functional Grade

Clone RMT1-10 is an anti-mouse TIM-1 (CD365) monoclonal antibody with a range of common in vivo applications in mice, focused primarily on immunomodulation, investigation of immune cell function, and disease model intervention.

Key established in vivo uses include:

• Attenuation of Atherosclerosis: RMT1-10 has been used to selectively expand peritoneal B1a cells (especially TIM-1+ IgM+ IL-10+ subsets) in mice, leading to a reduction in both the development and progression of atherosclerosis. This expansion enhances natural IgM antibody production and confers atheroprotective effects in models such as ApoE-knockout mice.

• Modulation of Autoimmune Disease Models: RMT1-10 treatment reduces the severity and delays the onset of experimental autoimmune encephalomyelitis (EAE), a mouse model for multiple sclerosis. It achieves this by promoting Th2 responses, inhibiting antigen-specific T cell proliferation, and altering cytokine production.

• Inhibition of T Cell Responses: In models of autoimmunity such as experimental allergic encephalomyelitis, RMT1-10 can inhibit antigen-specific T cell activation (e.g., PLP139-151-specific responses), impacting disease progression and immune tolerance.

• Promotion of Allograft Survival: RMT1-10 has been evaluated in transplantation models to prolong allograft survival, in part by modulating T cell responses and promoting regulatory immune mechanisms.

• Suppression of Atopic and Hypersensitivity Responses: The antibody is implicated in studies of airway hypersensitivity and atopic diseases, leveraging TIM-1's role as a co-stimulatory molecule for Th2 cell proliferation and function.

Mechanisms and Cell Targets:

• TIM-1 is expressed on activated CD4+ T cells (more on Th2 than Th1 cells) and B1a cells.
• RMT1-10 acts primarily as a blocking or non-agonistic antibody (rather than activating TIM-1 signaling), resulting in immune modulation rather than immune activation.

Application Summary Table:

Additional notes:

• RMT1-10 is typically administered intraperitoneally or intravenously in mice, with dosing and schedule tailored to the experimental design.
• Its effects are cell-type and context-dependent, sometimes suppressing or promoting particular immune functions.

These applications make RMT1-10 a versatile tool for dissecting the roles of TIM-1 in mouse models of immunity, inflammation, autoimmunity, and transplantation.

Commonly used antibodies and proteins studied alongside RMT1-10 (anti-TIM-1) in the literature are those that identify, quantify, or functionally characterize immune cell subtypes, particularly B1a cells and their markers, as well as markers relevant to the immune modulation effects of RMT1-10.

Frequently used antibodies and proteins include:

• CD19: Used to identify general B cells, including B1a cells, for flow cytometry analysis.
• CD5: Used in combination with CD19 to specifically gate B1a cells (CD19^+^CD5^+^).
• TIM-1: Used to detect TIM-1 expression on B1a and other immune cells, often with other anti-TIM-1 clones like 3B3, 3D10 for comparison or functional contrast.
• IgM: Marker for natural antibody-secreting B cells (B1a), often analyzed as IgM^+^ subpopulations.
• IL-10: Intracellular cytokine staining to assess regulatory B cells (B10 subset: IL-10^+^ B1a cells), as RMT1-10 treatment increases these populations.
• CD1d: Sometimes analyzed on B1a cells to further subset B cell populations after RMT1-10 treatment.
• Other anti-TIM-1 clones: For comparison, studies may use 3B3 or 3D10—these have different affinities and functional outcomes, often resulting in distinct immune modulation compared to RMT1-10.
• Isotype controls: Control antibodies of the same species and isotype as RMT1-10, to ensure specificity in flow cytometry or in vivo experiments.

On the protein side, other relevant molecules examined in parallel (via ELISA, immunostaining, or flow cytometry) may include:

• Cytokines such as IFN-γ, IL-4, and IL-17, as RMT1-10 affects T cell cytokine production and polarization.
• Lesion-associated proteins like oxidized LDL, VCAM-1, and MCP-1, in atherosclerosis models to evaluate downstream effects of RMT1-10.

RMT1-10 is almost always used in complex panels to characterize cell phenotype and function, particularly in the context of flow cytometry or immunohistochemistry, where multiple lineage and activation markers are evaluated in tandem.

In summary: Key antibodies and proteins frequently used with RMT1-10 encompass cell surface markers (CD19, CD5, CD1d), functional markers (IgM, IL-10), and occasionally other anti-TIM-1 clones (e.g., 3B3, 3D10), along with cytokines and lesion-related signals depending on disease model.

Clone RMT1-10 is an anti-mouse TIM-1 (T-cell immunoglobulin mucin domain 1) monoclonal antibody widely used to modulate immune responses in experimental models. The most consistently reported findings in the literature are:

• RMT1-10 promotes the induction and expansion of TIM-1⁺ regulatory B cells (Bregs), especially those secreting IL-10.
• RMT1-10 treatment provides protection in inflammatory models, including atherosclerosis, myocardial ischemia-reperfusion injury (IRI), and liver IRI.

Key findings from the scientific literature:

1. Atherosclerosis attenuation (in ApoE-KO mice on a high-fat diet):

   • RMT1-10 doubles or triples the number of TIM-1⁺ B1a cells in the peritoneal cavity and increases IgM-secreting and IL-10-producing B1a cells.
   • Atherosclerotic lesion size is reduced by 50%, with 40% and 38% reductions in lipid and macrophage accumulation.
   • Apoptotic cell numbers in lesions fall by 30%, and necrotic core size is reduced by almost 50%.
   • Efferocytosis (removal of apoptotic cells) improves by 81%.
   • These effects are linked to immune modulation rather than changes in plasma lipid levels or body weight.

2. Myocardial ischemia-reperfusion injury protection:

   • RMT1-10 reduces infarct (injury) size and apoptosis in heart tissue after IRI.
   • Increases TIM-1⁺ Bregs and IL-10 secretion.
   • Reduces inflammatory markers and improves cardiomyocyte morphology.
   • Activates Tregs (regulatory T cells) and inhibits pathogenic T cells after IRI.
   • The protective effects require B cells, as depletion by Anti-CD20 abolishes RMT1-10’s benefit.

3. Liver ischemia-reperfusion injury:

   • RMT1-10 alleviates liver injury and inflammation caused by IRI.
   • Increases TIM-1⁺ Bregs and promotes IL-10 production, supporting tissue protection and immune tolerance.

4. Mechanistic insights:

   • Effects are B cell-dependent, mediated primarily via expansion of functional TIM-1⁺ IL-10⁺ regulatory B cells.
   • RMT1-10 impacts classical immune regulatory pathways, shifting the balance toward anti-inflammatory and tolerogenic responses.
   • Does not directly affect overall B or T cell numbers except in specific subpopulations marked by TIM-1 expression.

5. Therapeutic implications:

   • Potential for RMT1-10 as a tool to investigate or therapeutically harness Breg-mediated anti-inflammatory effects in various immune-mediated diseases, transplant models, and tissue injury contexts.

In summary, clone RMT1-10 is widely cited for its role in expanding TIM-1⁺ regulatory B cells, enhancing IL-10-mediated immunoregulation, and decreasing tissue injury and inflammation in multiple disease models. The effects are highly dependent on Bregs, as depletion of B cells abolishes protection.

Dosing regimens of clone RMT1-10 (anti-mouse TIM-1 antibody) vary by mouse model, disease setting, and experimental endpoint, with differences in dose, frequency, and duration.

• Experimental Autoimmune Encephalomyelitis (EAE) Model (SJL Mice):

  • Example regimen: RMT1-10 was used at 10 μg/mL in cell culture for in vitro studies.
  • In vivo, RMT1-10 was shown to inhibit the development and severity of EAE when administered during the induction phase, though specific dosing details are not provided in the cited summary.

• Atherosclerosis Model (ApoE-KO Mice):

  • RMT1-10 was given intraperitoneally every other day for 8 weeks (prolonged regimen).
  • This approach contrasted with previous short-term experiments and was selected to assess long-term modulation of B cell compartments.

• Cardiac Allograft Transplantation (Multiple Strains):

  • Typical regimen: 0.5 mg (500 μg) intraperitoneally on day 0 (day of transplant), followed by 0.25 mg (250 μg) intraperitoneally on days 2, 4, 6, 8, and 10 after transplantation.
  • The regimen aims to promote allograft survival and modulation of immune responses in the acute phase post-transplant.

• Other Models/General Guidelines:

  • The antibody is typically administered intraperitoneally (i.p.).
  • Dose and schedule are tailored to disease context (acute vs. chronic immunomodulation) as well as mouse strain and age per standard immunology practices.

Observed Variations

Key points:

• Dose ranges from low μg (in vitro, mechanistic) to 250–500 μg per dose (in vivo transplant/chronic models).
• Regimen frequency adapts to experimental need: every other day for chronic models, or tightly clustered around a triggering event (e.g., transplantation).

In summary: RMT1-10 dosing regimens depend on mouse model, target disease, and experimental timeline, varying from single/short-term dosing around an induction event (e.g., transplantation or EAE onset) to long-term repeated administration (e.g., for chronic models like atherosclerosis). Always consult primary experiment methods for precise details under each model.