EL4 / EL4-luc Orthotopic Mouse Model Service for Lymphoma

The EL4 / EL4-luc orthotopic mouse model for lymphoma reproduces rapidly disseminating murine T-cell lymphoma in immunocompetent C57BL/6 hosts, combining the classic immunology workhorse with modern bioluminescence tracking of disease spread and survival. Committed exclusively to preclinical oncology services, Alfa Cytology delivers this syngeneic platform end to end: authenticated EL4 / EL4-luc cell supply, tail-vein orthotopic inoculation, scheduled in vivo imaging and survival monitoring, and terminal organ-level analysis of the lymphoid system.

Overview of EL4 / EL4-luc Orthotopic Mouse Model for Lymphoma

EL4 is a mouse T-cell lymphoma line with deep roots in immunology. Established from a lymphoma induced in a C57BL/6N mouse by the carcinogen 9,10-dimethyl-1,2-benzanthracene (DMBA), and later deposited from the Salk Institute’s Tumor Immunology Bank into ATCC as TIB-39, the line expresses the H-2^b and Thy-1.2 antigens, carries a modal chromosome number of 39, and displays the kind of quirks only a veteran line can: resistance to 0.1 mM cortisol, sensitivity to 20 µg/ml PHA, and reported low-level expression of the Gross murine leukemia virus antigen. EL4’s true fame, however, comes from its utility as an immunology tool: the subline EL4.IL-2 produces abundant interleukin-2, and the ovalbumin-transfected derivative E.G7-OVA is the standard model system for studying MHC class I-restricted cytotoxic T lymphocyte responses. Extended passage can alter the line’s immunogenicity, so early-passage, verified stocks are essential for reproducible in vivo work.

Fig 1. Subcutaneous EL4-Luc-GFP xenograft experiment in C57BL/6J mice showing that oral or intraperitoneal asparagine (Asn) administration significantly increased EL4-Luc-GFP tumor bioluminescence and tumor volume while aspartate (Asp) did not, validating the asparagine-dependency of EL4 tumors detected by BLI. (Deng, L., et al., 2020)

In orthotopic use, EL4 cells delivered intravenously colonize the lymphoid compartment of syngeneic C57BL/6 mice, producing aggressive, disseminated disease in a fully immunocompetent setting. A documented protocol injects 2 × 10⁶ luciferase-tagged EL4 cells via the tail vein into 6- to 8-week-old animals, with whole-animal bioluminescence captured biweekly after intraperitoneal firefly luciferin administration to chart longitudinal tumor progression; hind-limb paralysis serves as the clinical endpoint for survival analysis, typically reached within roughly two to three weeks of treatment initiation in published studies. Subcutaneous formats at the same inoculum grow palpable nodules within a week and support rapid pharmacology screens, while engineered derivatives—including EL4-Luc2 under the human EF1α promoter (ATCC TIB-39-LUC2), EL4-hCD20 bearing human CD20 for antibody and bispecific evaluation, and BCMA-luc EL4 for plasma cell-targeting concepts—extend the platform toward cell therapy and immunotherapy programs. Figure 1 outlines how this orthotopic lymphoma system is assembled and monitored.

Cell Line Information: EL4 / EL4-luc

Few mouse lines have served immunology research as long or as faithfully as EL4. The defining characteristics of the parent line and its luciferase-tagged derivatives are summarized below.

Our Services

An EL4 / EL4-luc engagement with Alfa Cytology runs as one continuous pipeline: early-passage verified cell banking, standardized tail-vein orthotopic inoculation, biweekly IVIS sessions aligned with the disease timeline, dosing per the client’s regimen, and a terminal package combining survival statistics, ex vivo organ imaging, and lymphoid-system flow cytometry.

Workflow of EL4 / EL4-luc Orthotopic Mouse Model Construction

Deploying the EL4 / EL4-luc orthotopic lymphoma model is a compact, fast-turnaround workflow in which small intravenous inocula reliably generate disseminated disease within days. The workflow below reflects the standard execution of this platform.

1. Cell Bank Verification and Expansion: EL4 or EL4-luc stocks are authenticated, mycoplasma-tested, and expanded from early passages; for immunotherapy applications, passage number is logged and drift-sensitive stocks are used.
2. Luciferase Calibration (EL4-luc Only): Signal output per cell is verified in vitro so that in vivo photon flux can be converted into relative tumor burden across cohorts.
3. Mouse Randomization and Grouping: Six- to eight-week-old C57BL/6 mice are acclimatized and randomized by body weight under approved animal protocols, with no immunodeficiency required for syngeneic engraftment.
4. Orthotopic Tail-Vein Inoculation: A single-cell suspension of 2 × 10⁶ EL4 or EL4-luc cells in a small volume of sterile buffer is delivered via the lateral tail vein so that cells seed the lymphoid compartment and marrow.
5. Engraftment Confirmation and Baseline Imaging: Whole-animal bioluminescence after intraperitoneal luciferin confirms cell take within days and establishes baseline radiance per animal.
6. Treatment Administration: Test agents, immunotherapy combinations, or targeted nanocarriers are dosed per the client’s schedule, with dosing typically initiated around day 9 after inoculation.
7. Longitudinal Monitoring and Survival Endpoint: Biweekly IVIS imaging tracks dissemination while body weight and clinical status are recorded daily; hind-limb paralysis or pre-set welfare criteria define the survival endpoint for Kaplan–Meier analysis.
8. Terminal Organ Analysis: At endpoint, thymus, spleen, lymph nodes, bone marrow, and peripheral blood are collected for ex vivo bioluminescence, flow cytometric profiling of tumor-infiltrating immune populations (e.g., Treg frequencies within CD45⁺ cells), and histopathology.

Fig 2. EL4 / EL4-luc orthotopic mouse model construction workflow.

Case Study-EL4 / EL4-luc Orthotopic Mouse Model Development

Alfa Cytology has executed preclinical programs on the EL4 / EL4-luc orthotopic platform for clients benchmarking immunotherapy combinations, validating bispecific and CAR-based constructs against humanized targets, and profiling nanoparticle delivery systems, delivering longitudinal bioluminescence records, survival analyses, and tumor microenvironment flow data. Because each engagement is configured around the client’s own therapeutic concept, study outlines and representative datasets are available upon request.

Why Choose Alfa Cytology?

EL4 / EL4-luc orthotopic work is fast-moving and passage-sensitive: drifted stocks or mistimed imaging can silently distort immunotherapy results. Alfa Cytology’s stock management and timeline discipline keep these syngeneic lymphoma studies tight, reproducible, and decision-ready.

• Early-passage, authenticated EL4 / EL4-luc banks with mycoplasma verification and drift monitoring.
• Standardized tail-vein orthotopic inoculation delivering reliable disseminated disease in immunocompetent hosts.
• Calibrated in-house IVIS imaging aligned to the model’s compressed timeline.
• Syngeneic immunocompetent setting preserving full immune-tumor interactions for immunotherapy evaluation.
• Survival-based endpoints with lymphoid-organ ex vivo imaging and microenvironment flow cytometry.
• Rapid study turnaround thanks to the model’s short disease course and experienced execution team.

Contact Us

Whether the goal is to test an immunotherapy combination, validate a bispecific construct, or track disseminated lymphoma in real time, Alfa Cytology can stand up an EL4 / EL4-luc orthotopic study on your timeline. Contact us today to scope your study design and dosing regimen, or reach out to our team for a customized quotation.

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