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TIM3 Humanized Mouse Models

TIM3 humanized mice enable preclinical testing of anti human TIM3 therapeutic antibodies in immunocompetent mice for next generation checkpoint immunotherapy development.

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TIM3 in Cancer Immunotherapy

T cell immunoglobulin and mucin domain containing 3 (TIM3, also known as HAVCR2) is an immune checkpoint receptor under active clinical development, with multiple anti TIM3 antibodies in trials for various malignancies.

TIM3 Biology

TIM3 is expressed on multiple immune cell populations and contributes to immune regulation:

T Cell Expression

TIM3 is upregulated on activated and exhausted T cells. Co expression of TIM3 with PD1 marks the most dysfunctional T cell population in chronic infection and cancer.

Innate Immune Cell Expression

TIM3 is constitutively expressed on macrophages, dendritic cells, and mast cells, regulating their inflammatory responses.

NK Cell Expression

TIM3 is expressed on natural killer cells and can inhibit their cytotoxic function.

TIM3 Ligands

TIM3 has multiple ligands affecting its function:

LigandFunction
Galectin 9Carbohydrate binding protein that triggers TIM3 mediated T cell death
PhosphatidylserineTIM3 binds phosphatidylserine on apoptotic cells, contributing to clearance
CEACAM1Carcinoembryonic antigen related cell adhesion molecule 1 binds TIM3 and promotes T cell exhaustion
HMGB1High mobility group box 1 binding to TIM3 on dendritic cells interferes with nucleic acid sensing

Clinical Anti TIM3 Antibodies

Multiple anti TIM3 antibodies are in clinical development:

Cobolimab (TSR 022)

Anti TIM3 antibody in combination trials with anti PD1

Sabatolimab (MBG453)

Anti TIM3 antibody being evaluated in hematologic malignancies and solid tumors

Additional Candidates

Multiple other anti TIM3 antibodies are in early clinical development

Species Specificity

Clinical anti TIM3 antibodies are designed for human TIM3 and typically do not cross react with mouse Tim3, necessitating humanized models for preclinical evaluation.

TIM3 Humanization Strategies

Complete Gene Replacement

Full replacement of mouse Havcr2 with human HAVCR2 (TIM3) coding sequences:

Endogenous Regulatory Control

Human TIM3 expressed from the native mouse promoter maintains physiological expression patterns.

Functional Preservation

The humanized allele preserves TIM3 function on T cells, NK cells, and myeloid populations.

Extracellular Domain Humanization

Humanization of the extracellular domain provides human epitopes for antibody binding.

Design Considerations

Expression on Multiple Cell Types

TIM3 is expressed on T cells, NK cells, dendritic cells, and macrophages. Humanization should preserve expression across these populations.

Ligand Binding

Consider whether human TIM3 appropriately binds mouse galectin 9 and other ligands.

Signaling Compatibility

Mouse intracellular domains may be retained for compatibility with mouse signaling machinery.

Applications of TIM3 Humanized Mice

Monotherapy Evaluation

  • Syngeneic Tumor Models:TIM3 humanized mice combined with syngeneic tumors enable efficacy assessment
  • Dose Response:Establish pharmacologically relevant dosing for clinical candidates
  • Target Engagement:Measure antibody binding to TIM3 positive cells in tumor and lymphoid tissues

Combination Checkpoint Blockade

  • TIM3 plus PD1:The most common combination under investigation. Dual humanized models expressing both human TIM3 and human PD1 enable testing
  • TIM3 plus LAG3:Alternative checkpoint combinations
  • Triple Combinations:TIM3 plus PD1 plus CTLA4 or other three way combinations

T Cell Exhaustion Studies

  • Exhaustion Reversal:Assess whether TIM3 blockade reinvigorates exhausted T cells
  • Combination Effects:Evaluate synergy with PD1 blockade in reversing exhaustion
  • Phenotypic Analysis:Characterize changes in exhaustion markers following TIM3 blockade

Multi Checkpoint Humanized Models

Combination checkpoint approaches require models expressing multiple human targets:

Dual Humanized Models

TIM3 plus PD1Both checkpoints humanized for testing emerging combinations
TIM3 plus LAG3For alternative combination studies

Higher Order Combinations

TIM3 plus PD1 plus LAG3Triple humanization for comprehensive checkpoint blockade studies
TIM3 plus PD1 plus CTLA4Alternative triple combination

Model Design Considerations

Background Strain Selection

C57BL/6

Standard background for immuno oncology studies. Compatible with commonly used syngeneic tumor models.

BALB/c

Alternative for specific tumor models.

C57BL/6 Mouse Background

Syngeneic Tumor Compatibility

TIM3 humanized mice should be crossed to appropriate syngeneic compatible backgrounds:

MC38 Colon CarcinomaCommonly used for checkpoint studies
B16 MelanomaRelevant for melanoma immunotherapy
CT26 Colon CarcinomaBALB/c compatible option

Expression Validation

Before experimental use, confirm:

Surface Expression

Flow cytometry for human TIM3 on T cells, NK cells, and myeloid populations

Inducible Expression

TIM3 upregulation following T cell activation

Antibody Binding

Clinical candidates bind to humanized TIM3

Phenotyping TIM3 Humanized Models

Immune Characterization

T Cell Populations

Flow cytometry for CD4, CD8 T cells with TIM3 and PD1 co staining

Exhaustion Phenotype

TIM3/PD1 double positive cells represent exhausted population

NK Cell Expression

TIM3 on NK cells

Myeloid Expression

TIM3 on dendritic cells and macrophages

Our Approach to TIM3 Humanized Models

Quality Assurance

ingenious targeting laboratory validates TIM3 humanized models:

  • Flow Cytometry: Confirms human TIM3 surface expression
  • Functional Testing: Binding of clinical antibody candidates
  • Baseline Immune Status: Normal immune development and function

Selected Publications in TIM3 Research

According to PubMed, recent publications demonstrate the importance of TIM3 in checkpoint immunotherapy:

Chen C et al. (2024)

Soluble Tim 3 serves as a tumor prognostic marker and therapeutic target for CD8 T cell exhaustion and anti PD1 resistance

Cell Reports Medicine

Soltantoyeh T et al. (2024)

Simultaneous targeting of Tim3 and A2a receptors modulates MSLN CAR T cell antitumor function in a human cervical tumor xenograft model

Frontiers in Immunology

View All Publications

What Researchers Say

iTL produced a new conditional mouse model for us and the quality of service was exceptional. The team is extremely knowledgeable and the work was completed at the highest possible standards. My project manager was excellent and always happy to answer technical questions and keep me up to date with progress and potential problems. I would recommend iTL highly and will use them again in the future if I need to generate a new mouse line.

Albert Basson, PhD

King's College London

✦ New for 2026

Breeding Scheme Architect

Plan complex multi-allele breeding strategies, calculate expected genotype ratios, and estimate time to experimental cohorts—all before starting your project.

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Allele 1Gene-flox (conditional)
Allele 2Cre-driver (tissue-specific)
TargetHomozygous knockout

→ 3 generations to target genotype

Start Your TIM3 Project

Ready to discuss TIM3 humanization for your immuno oncology research? Our scientific team provides complimentary consultation to design the optimal model for your program.

Frequently Asked Questions

Full humanization replaces the entire mouse TIM3 (Havcr2) gene with human TIM3 (HAVCR2) sequence. ECD only humanization replaces only the extracellular domain while retaining mouse transmembrane and cytoplasmic domains. Full humanization is preferred for antibody testing; ECD only may preserve some mouse signaling functions.

Yes. TIM3 humanization can be combined with PD1, PDL1, CTLA4, LAG3, or other checkpoint humanizations to create double, triple, or multi checkpoint models. Common combinations include TIM3+PD1 and TIM3+LAG3 for combination checkpoint blockade studies targeting exhausted T cells.

Yes. TIM3 humanized mice can be combined with syngeneic tumor cell lines (MC38, B16, LLC) to create systems where both tumor and immune compartments express human targets. This enables evaluation of checkpoint blockade in immunocompetent animals with intact tumor immunity, providing more physiologically relevant testing.

Lab Signals

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