E.G7-OVA / E.G7-OVA-luc Orthotopic Mouse Model Service for Lymphoma

The E.G7-OVA / E.G7-OVA-luc orthotopic mouse model of lymphoma combines a defined tumor antigen — the SIINFEKL epitope of chicken ovalbumin presented on H-2Kb — with an intact syngeneic immune system, making it possible to track antigen-specific T-cell immunity and therapeutic response quantitatively in vivo. Alfa Cytology delivers this platform as a complete preclinical service: authenticated EL4/E.G7-OVA cell stewardship under G418 selection, luciferase line qualification, engraftment in immunocompetent C57BL/6 recipients, scheduled whole-body bioluminescence imaging, checkpoint-blockade and adoptive-cell-therapy dosing, and immune-correlative endpoint analysis.

Overview of E.G7-OVA / E.G7-OVA-luc Orthotopic Mouse Model for Lymphoma

E.G7-OVA descends from EL4, a T-cell lymphoma line induced in a C57BL mouse by the carcinogen 9,10-dimethyl-1,2-benzanthracene, and represents one of immunology’s most consequential engineering achievements: in 1988, Moore and colleagues stably transfected EL4 with the full-length chicken ovalbumin gene using the pAc-neo-OVA plasmid, creating a tumor clone that naturally processes and presents the OVA 257-264 peptide (SIINFEKL) in the context of H-2Kb — measured at roughly 90 peptide-MHC complexes per cell. Because ovalbumin is foreign to mice, E.G7-OVA converts a poorly immunogenic, self-antigen-only lymphoma into a model of a strongly antigenic tumor, while its H-2b, B7-negative background keeps antigen presentation clean and interpretable. The line is PD-L1 positive, and a PD-L1-overexpressing derivative (E.G7-PD-L1) extends the system for ligand-density studies.

Fig 1. Therapeutic E.G7-OVA tumor model in C57BL/6 mice (tumor challenge day 0, nanovaccine days 6/10/14) showing that BN@HM-OVA nanovaccine nearly eradicated E.G7-OVA tumors with 20-day growth curves, dissected tumor weights and body weight tracking, outperforming HM-OVA, N@HM-OVA and other formulations. (Xie, Xiaoxue, et al., 2022)

In vivo, E.G7-OVA cells inoculated into syngeneic C57BL/6 mice develop progressive, dose-dependent solid tumors that typically kill untreated recipients within about four weeks — and yet the model’s true power is immunological: paired with OT-I TCR-transgenic T cells (Vα2/Vβ8.1 receptors specific for SIINFEKL-H-2Kb) it forms a complete antigen-recognition toolkit in which tumor-specific CD8+ responses can be quantified, transferred, and therapeutically manipulated. Luciferase tagging of the EL4 lineage enables non-invasive bioluminescence tracking of tumor burden on imaging platforms, adding a quantitative dimension to a model classically read by caliper. The platform has generated landmark findings across checkpoint blockade — where anti-PD-L1 monotherapy and cisplatin-plus-anti-PD-L1 combinations show antitumor activity that is strictly antigen-dependent (parental EL4, lacking OVA, is refractory) — adoptive T-cell therapy with engineered OT-I cells, GM-CSF-secreting vaccine plus CTLA-4 blockade combinations, anti-VISTA programs, and CAR-T/cytokine-enhanced cell therapy studies, making E.G7-OVA-luc the reference syngeneic system for antigen-defined cancer immunotherapy research.

Cell Line Information: E.G7-OVA / E.G7-OVA-luc

E.G7-OVA is an engineered murine lymphoma line whose precisely defined antigen biology underpins decades of immunotherapy research; its luciferase derivative extends it into imaging-guided in vivo studies. Core attributes follow.

Our Services

An antigen-defined model is only as trustworthy as the antigen it presents — drift in ovalbumin expression or loss of G418 selection quietly invalidates results. Alfa Cytology therefore runs the E.G7-OVA platform with unusual stringency: master banks are maintained under continuous selection, expression of the SIINFEKL-H-2Kb complex is verified before release, and luciferase lots are qualified for uniform in vitro glow. Engraftment, imaging, and checkpoint or adoptive-cell dosing follow written schedules, and immune-correlative readouts — from draining-lymph-node cytolytic assays to tumor-infiltrate profiling — close out every cohort. Across the full E.G7-OVA / E.G7-OVA-luc orthotopic workflow, Alfa Cytology gives immunology-focused sponsors one accountable partner.

Workflow of E.G7-OVA / E.G7-OVA-luc Orthotopic Mouse Model Construction

The E.G7-OVA-luc workflow is built for immunological interpretability: antigen expression is verified before a single animal is enrolled, dosing schedules mirror the published checkpoint and adoptive-therapy literature, and endpoints connect tumor control to antigen-specific immunity. The sequence below is the standard flow.

1. Line stewardship and antigen verification: E.G7-OVA / E.G7-OVA-luc stocks are expanded at low passage under continuous G418 selection, mycoplasma-tested, and released only after confirming SIINFEKL-H-2Kb surface presentation and uniform luciferase expression by in vitro assay.
2. Inoculum preparation: Cultures maintained at 0.1-1.0×10⁶ cells/mL are harvested at log phase, washed, counted, and resuspended in sterile PBS at the study dose (commonly 2×10⁵ to 2×10⁶ cells in 100 µL for subcutaneous engraftment).
3. Syngeneic recipient preparation: Immunocompetent C57BL/6 mice are acclimatized and body-weight stratified; where designs require, OT-I or other TCR-transgenic donor mice are staged in parallel for effector-cell preparation.
4. Orthotopic engraftment and baseline imaging: The suspension is injected subcutaneously into the flank tumor-bed (or via the tail vein for systemic designs), and baseline whole-animal bioluminescence after D-luciferin confirms engraftment and starting burden.
5. Treatment-group allocation: Once tumors reach defined size, mice are randomized into arms — checkpoint antibodies (anti-PD-L1, anti-CTLA-4, anti-VISTA classes), chemotherapy comparators such as cisplatin, vaccines, or vehicle — with parental EL4 arms available as antigen-negative controls.
6. Adoptive-cell dosing and longitudinal monitoring: Where designed, OT-I or engineered T cells (for example, 2×10⁶ cells intravenously around day 7 post-tumor) are transferred, serial bioluminescence and caliper measurements track tumor growth twice weekly, and body weight and clinical condition are scored throughout.
7. Immune-correlative endpoints: At study end, tumors, draining lymph nodes, spleens, and blood are collected for tumor weight, ex vivo imaging, cytolytic assays against E.G7-OVA versus EL4 targets, and flow-cytometric profiling of tumor-infiltrating lymphocytes, with histopathology confirming lymphoma involvement.

Fig 2. E.G7-OVA / E.G7-OVA-luc orthotopic mouse model construction workflow.

Case Study-E.G7-OVA / E.G7-OVA-luc Orthotopic Mouse Model Development

Alfa Cytology has implemented the E.G7-OVA-luc platform in preclinical programs spanning checkpoint inhibitors, combination chemo-immunotherapy, and adoptive cell therapies. In these studies, longitudinal bioluminescence paired with caliper tracking resolved treatment-induced growth control against matched baselines, parental EL4 antigen-negative arms confirmed antigen dependence of response where designed, and OT-I transfer schedules mirrored the published checkpoint-synergy literature. Endpoints connected tumor outcomes to mechanism through draining-lymph-node cytolytic assays and tumor-infiltrating-lymphocyte flow-cytometric profiling. Program-specific designs and results are retained as sponsor-confidential deliverables, and representative outlines can be furnished upon request.

Why Choose Alfa Cytology?

In an antigen-defined model, the controls are the experiment — Alfa Cytology’s E.G7-OVA-luc service is engineered around that principle. Sponsors choosing this platform gain:

• Continuous G418-maintained master banks with verified SIINFEKL-H-2Kb presentation before study release.
• Luciferase lines qualified for uniform in vitro glow output, supporting quantitative whole-animal imaging.
• Syngeneic immunocompetent C57BL/6 hosts preserving full tumor-immunity interactions.
• Built-in antigen-dependence logic using parental EL4 arms as negative controls.
• Integrated OT-I sourcing and adoptive-transfer scheduling aligned with published checkpoint-synergy designs.
• Dual tumor readouts combining bioluminescence with caliper measurements for cross-validation.
• Mechanistic endpoint packages spanning cytolytic assays and tumor-infiltrating-lymphocyte profiling.

Contact Us

Designing an antigen-defined immunotherapy study and need a rigorously controlled syngeneic lymphoma platform? Contact us to discuss engraftment dose, checkpoint or adoptive-therapy schedules, and correlative endpoints for the E.G7-OVA / E.G7-OVA-luc orthotopic model. Alternatively, reach out to Alfa Cytology for a customized proposal and quotation — our immunology-aware preclinical team will respond with a design built around your mechanism.

For research use only. Not intended for any clinical use.