E.G7-OVA Syngeneic Model Service for Lymphoma

The E.G7-OVA syngeneic model of lymphoma pairs the historic EL4 T-cell lymphoma with a single-copy, constitutively secreted ovalbumin cassette, creating immunocompetent C57BL/6 hosts in which tumor antigen is abundant, defined and traceable.Alfa Cytology delivers the complete E.G7-OVA workflow—authenticated cell banking, flank implantation, OT-I/OT-II-compatible designs, checkpoint and costimulatory intervention schedules, and tetramer-based endpoint analysis—giving sponsors decision-ready preclinical datasets shaped to their program, strictly within a preclinical scope.

Overview of E.G7-OVA Syngeneic Model for Lymphoma

E.G7-OVA’s pedigree is written into its accession numbers. The parental line, EL4 (ATCC TIB-39), is a T lymphoblast established from a 9,10-dimethylbenzanthracene-induced lymphoma in a C57BL mouse—cortisol-resistant, phytohemagglutinin-sensitive, bearing Thy-1.2 and H-2^b antigens with a modal chromosome number of 39—and it has an immunological family tree all its own, including an IL-2-hyperproducing subline (EL4.IL-2, ATCC TIB-181) and a celebrated series of costimulatory transfectants. In 1988, Michael Bevan’s group converted this workhorse into an antigen-defined instrument: EL4 was electroporated with the plasmid pAc-neo-OVA, which carries full-length chicken ovalbumin cDNA together with neomycin resistance, and clone E.G7—harboring exactly one inserted copy—became E.G7-OVA (ATCC CRL-2113). The cells constitutively synthesize and secrete OVA, which means antigen is not merely expressed but released into the tumor microenvironment—feeding cross-presentation by host dendritic cells and making the line, in ATCC’s own description, a model system for studying MHC class I–restricted cytotoxic T-lymphocyte responses. Immunized C57BL/6 mice mount H-2K^b-restricted CTLs against the dominant OVA epitopes, and the line’s T-cell identity (CD3⁺, CD4⁻, CD8⁻) has made it a fixture of lymphoma immunology.

Fig 1. Anti-PD-L1 efficacy in the E.G7 syngeneic lymphoma model — individual tumor growth curves showing 0/11 tumor-free in the rat IgG2a control group versus 7/13 complete regressions in the anti-PD-L1 group, mean tumor volumes with significant reduction (**P<0.0001 at day 22), and overall survival (percent of E.G7 tumor-bearing mice over 25 days, P<0.01), establishing the E.G7 model as responsive to PD-L1 blockade via host-cell PD-L1. (Rodriguez-Barbosa, Jose-Ignacio, et al., 2020)

In vivo, E.G7-OVA is gratifyingly economical: half a million cells in the flank—one-sixth of the parental EL4 inoculum—produce uniform, brisk tumors, tracked by the classic (short diameter)² × long diameter ÷ 2 formula to a 2,000 mm³ survival endpoint. Its second identity is as a testing ground for immunomodulatory antibodies: the EL4 backbone carried B7-1 and B7-2 transfectants whose study revealed interleukin-4–dependent immune suppression by B7-2, and E.G7-OVA tumors have since been used to benchmark agonistic anti-4-1BB, anti-OX40 and anti-GITR antibodies. Contemporary usage keeps expanding—6-phosphogluconate dehydrogenase blockade was shown to invigorate CD8 effector function against E.G7-OVA, the histone demethylase Utx was linked to CD8-dependent antitumor control in the same model, ferroptotic cell death was interrogated for its immunosuppressive consequences here, and invariant-NKT-cell in situ vaccination has been prototyped against these tumors. Discipline items: G418 selection (0.4–0.5 mg/mL) is required to maintain OVA expression, the double-negative T-cell phenotype invites caution in subset-level interpretations, brisk growth compresses windows, and—as with all OVA systems—the model antigen carries a translation gap.

Cell Line Information: E.G7-OVA

Key characteristics of E.G7-OVA and its parental heritage are summarized below.

Our Services

With in vivo oncology and T-cell-biology specialists guiding each stage and flow cytometry, tetramer staining and digital pathology in-house, Alfa Cytology configures every E.G7-OVA engagement around the sponsor’s antigen-specific question—controlling OVA expression through enforced G418 selection, coordinating OT-I/OT-II logistics, and pairing interventions with tetramer-based readouts for clean, decision-grade preclinical data.

Workflow of E.G7-OVA Syngeneic Model Construction

Construction of the E.G7-OVA syngeneic model is an implantation workflow whose quality rests on verified antigen expression, vigorous C57BL/6 recipients and disciplined monitoring. A standard engagement proceeds as follows:

1. Source and qualify cells: E.G7-OVA stocks are cleared by mycoplasma PCR and identity-verified against the published profile, with OVA expression confirmed by Western blot and SIINFEKL–H-2K^b staining (25-D1.16); G418 selection is maintained through expansion, and luciferase labeling is titrated in vitro where imaging readouts are required.
2. Prepare recipients: C57BL/6 mice (typically 6–8 weeks old; OT-I/OT-II transgenic or congenically marked hosts where antigen-specific designs call for them) complete acclimation and health screening.
3. Prepare the inoculum: log-phase suspension cells are collected, washed to remove residual secreted OVA and selection drug, counted, checked for viability and resuspended in PBS (5 × 10⁵ cells in 100 µL per flank).
4. Implant: cells are injected subcutaneously into the flank with consistent anatomical positioning for reproducible caliper measurement.
5. Randomize: when tumors reach palpability (typically within the first week), animals are assigned to groups balanced by tumor volume and body weight.
6. Dose and follow: test articles (checkpoint or costimulatory agents, vaccines, metabolic inhibitors) are administered on schedule; tumors are measured two to three times weekly using the (short diameter)² × long diameter ÷ 2 formula, with body weight and survival logged to the 2,000 mm³ endpoint.
7. Terminate and analyze: tumors are resected and weighed; draining lymph nodes, spleen and lungs are examined; tumor-infiltrating lymphocytes and splenocytes are analyzed by flow cytometry with H-2K^b–SIINFEKL tetramers and activation/exhaustion panels; sections are stained with 25-D1.16 and CD3; tissues enter histopathology and molecular analysis.

Fig 2. E.G7-OVA Syngeneic Model construction workflow.

Case Study-E.G7-OVA Syngeneic Model Development

Alfa Cytology maintains an active preclinical case-study file on the E.G7-OVA platform: establishment-phase verification of OVA expression stability under G418 selection, in vivo growth kinetics at the 5 × 10⁵-cell inoculum, costimulatory- and checkpoint-based intervention studies with antigen-specific monitoring, and endpoint analyses combining tetramer staining, 25-D1.16 immunohistochemistry and TIL phenotyping. Representative growth curves, T-cell tracking datasets and endpoint summaries can be shared upon request and will be updated as new programs conclude.

Why Choose Alfa Cytology?

E.G7-OVA’s secreted-antigen design and low inoculum make it unforgiving of sloppy culture and rewarding of precise execution—which is where Alfa Cytology invests:

• G418-enforced banking with Western blot and 25-D1.16 verification before every implantation.
• Low-inoculum expertise (5 × 10⁵ cells) with uniform, brisk take as a routine outcome.
• Costimulatory-antibody testing experience reflecting the model’s documented 4-1BB, OX40 and GITR pedigree.
• Cross-presentation-friendly designs exploiting the secreted-OVA biology.
• In-house tetramer staining and multiparameter flow cytometry for antigen-specific TIL quantification.
• Brisk-kinetics scheduling so compressed windows are anticipated and planned.
• One accountable project contact with milestone-by-milestone visibility.
• A strictly preclinical scope—every study is planned, executed and reported as non-clinical research.

Contact Us

Interested in the E.G7-OVA syngeneic lymphoma model for your next preclinical program? Reach out to us with your antigen-specific design, timeline and endpoint wish-list, and the Alfa Cytology team will respond with a customized plan covering model setup, OT-I/OT-II coordination and analysis workflow—contact us today and move your program forward with confidence.

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