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Checkpoint Humanization & Tumor Immunity

Immuno-Oncology Mouse Models

Since 1998, ingenious targeting laboratory has supported immuno-oncology researchers with custom mouse models contributing to peer reviewed publications in Cancer Research, Nature Medicine, Immunity, and leading oncology journals worldwide.

Immuno oncology mouse models enable researchers to investigate the complex interactions between tumors and the immune system, from T cell exhaustion and checkpoint regulation to tumor microenvironment dynamics. Humanized immune checkpoint models express human versions of PD1, PDL1, CTLA4, and other targets, allowing direct testing of clinical antibody candidates in immunocompetent mice.

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Humanized Immune Checkpoint Models

Why Humanize Checkpoints

Many therapeutic antibodies targeting immune checkpoints are designed specifically for human proteins and show limited or no cross reactivity with mouse orthologs. Humanized checkpoint models express human versions of these targets, enabling:

Direct testing of clinical antibody candidates
Evaluation of antibody effector functions
Combination therapy optimization
Biomarker development and validation
Pharmacokinetic and pharmacodynamic studies

Single vs Dual Humanization

Single humanization

One checkpoint target humanized for monotherapy studies

Dual humanization

Two checkpoints humanized (e.g., PD1 + CTLA4) for combination studies

Multi target

Three or more checkpoints for complex combination evaluation

Available Checkpoint Targets

TargetFunctionTherapeutic Applications
PD1 (PDCD1)T cell inhibitory receptorAnti PD1 antibody efficacy
PDL1 (CD274)PD1 ligand on tumor/immune cellsAnti PDL1 antibody efficacy
CTLA4T cell inhibitory receptorAnti CTLA4 efficacy, combinations
LAG3T cell exhaustion markerLAG3 inhibitor evaluation
TIM3 (HAVCR2)T cell exhaustion markerTIM3 pathway studies
TIGITNK and T cell inhibitory receptorTIGIT inhibitor development
GITRCostimulatory receptorAgonist antibody evaluation
OX40Costimulatory receptorAgonist antibody evaluation
4-1BB (CD137)Costimulatory receptorAgonist antibody evaluation

Genetic Tumor Models

Tumor Suppressor Knockouts

Conditional knockout of tumor suppressors enables controlled tumor development in immunocompetent hosts:

p53 conditional knockout:Spontaneous tumor development in specific tissues
PTEN conditional knockout:PI3K pathway activation, various tumor types
APC conditional knockout:Intestinal tumor models
Rb conditional knockout:Cell cycle dysregulation

Oncogene Activation Models

Conditional expression of activated oncogenes enables controlled tumor initiation:

KRAS G12D knockin:Lung and pancreatic tumor models
BRAF V600E knockin:Melanoma and other tumor types
MYC overexpression:Various tumor types
Fusion oncogenes:Chromosomal translocation models

Combining Genetic Models with Checkpoint Humanization

Genetic tumor models can be combined with humanized checkpoint alleles to create sophisticated platforms for immunotherapy evaluation: autochthonous tumors in checkpoint humanized hosts, spontaneous tumor development with human checkpoint targets, and physiologically relevant tumor microenvironment.

Syngeneic Tumor Models

Syngeneic tumor cells transplanted into checkpoint humanized mice enable rapid evaluation of clinical antibody candidates:

Cell LineTumor TypeBackgroundCommon Applications
MC38Colon carcinomaC57BL/6Checkpoint inhibitor efficacy
CT26Colon carcinomaBALB/cImmunotherapy combinations
B16MelanomaC57BL/6Immunotherapy, checkpoint studies
LLCLung carcinomaC57BL/6Metastasis, immunotherapy
4T1Breast carcinomaBALB/cMetastatic disease, immunotherapy
EMT6Breast carcinomaBALB/cImmunotherapy response

Model Types for Immuno-Oncology

Humanized Knockin Models

Humanized checkpoint knockins replace mouse genes with human orthologs at the endogenous locus:

  • Physiological expression levels
  • Normal tissue distribution
  • Endogenous regulatory control
  • Maintained immune system function
Learn more

Conditional Knockout Models

Conditional knockouts enable tissue specific or temporal gene manipulation:

  • Immune cell specific deletions (T cell, B cell, myeloid)
  • Tumor cell specific deletions
  • Inducible knockouts modeling therapeutic intervention
  • Target validation through genetic ablation
Learn more

Reporter Models

Reporter knockins enable visualization and tracking of immune cells:

  • Fluorescent reporters for immune cell tracking
  • Luciferase reporters for in vivo imaging
  • Lineage tracing of tumor infiltrating lymphocytes
  • Real time monitoring of immune activation
Learn more

Research Applications

Therapeutic Antibody Development

  • Anti checkpoint antibody efficacy testing
  • Agonist antibody evaluation
  • Antibody effector function studies
  • Combination therapy optimization
  • Dose response and scheduling studies

Mechanism of Action Studies

  • T cell exhaustion and reinvigoration
  • Tumor microenvironment dynamics
  • Resistance mechanism identification
  • Biomarker discovery and validation

Combination Immunotherapy

  • Checkpoint combinations (PD1 + CTLA4)
  • Checkpoint plus chemotherapy
  • Checkpoint plus targeted therapy
  • Checkpoint plus radiation

Selected Publications

Models generated by ingenious targeting laboratory have supported immuno oncology research:

Clausen BE et al. (1999).

Conditional gene targeting in macrophages and granulocytes using LysMcre mice.

Transgenic Research 8(4): 265-277

Chakrabarti S et al. (2024).

Touch sensation requires the mechanically gated ion channel ELKIN1.

Science 383(6686): 992-998

What Researchers Say

I'd like to thank the ingenious team for making this mouse for us. We are so excited! Everyone at ingenious has been wonderful to work with throughout the entire process. We will definitely be in contact the next time we need a mouse!

Julia Maxson, PhD

Knight Cancer Institute, Oregon Health & Science University

Start Your Immuno-Oncology Project

Our scientific consultants are ready to discuss your immuno oncology research requirements and recommend the optimal model design for your therapeutic program. Initial consultation is provided at no charge and includes target analysis, checkpoint humanization strategy, and timeline estimates.

✦ 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.

Visualize multi-generation breeding paths
Calculate Mendelian ratios automatically
Estimate timeline to study-ready cohorts

Free Research Tool

No account required

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

→ 3 generations to target genotype

Frequently Asked Questions

Most checkpoint inhibitor antibodies target human proteins and do not cross-react with mouse orthologs. Humanized checkpoint models express human PD1, PDL1, CTLA4, or other targets, enabling efficacy testing of clinical antibodies in immunocompetent mice with functional immune systems. This is essential for preclinical immunotherapy development.

Single checkpoint models (e.g., PD1 humanized) enable testing of monotherapy antibodies. Dual checkpoint models (e.g., PD1/CTLA4 humanized) enable testing of combination immunotherapy, which is increasingly important in clinical practice. Dual humanization requires combining multiple humanized alleles through breeding.

Yes. Humanized checkpoint models on C57BL/6 or BALB/c backgrounds are compatible with syngeneic tumor lines (MC38, B16, CT26, 4T1). This enables testing of checkpoint inhibitors in immunocompetent mice with functional immune systems, providing more physiologically relevant data than immunodeficient models.

GEMMs develop tumors spontaneously from defined genetic alterations and better recapitulate tumor evolution and microenvironment. Syngeneic models involve injecting established tumor cell lines and are faster for screening studies. Both approaches have value: GEMMs for mechanism studies and disease modeling, syngeneic models for rapid immunotherapy screening.

C57BL/6 is most common for Th1-biased immune responses and compatibility with MC38 and B16 syngeneic tumors. BALB/c is preferred for Th2-biased responses and CT26/4T1 tumors. Strain selection affects immune phenotype and tumor growth, so choose based on your experimental requirements and syngeneic tumor compatibility.

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Immuno-Oncology Research Insights

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