In Vivo CAR-T Cell Functional Analysis Service after Amphi-ligand Vaccination

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All products and services are For Research Use Only and CANNOT be used in the treatment or diagnosis of disease.

The study revealed a novel strategy to utilize CAR T to treat antigenically heterogeneous solid tumors, i.e., to enhance IFN-γ production through vaccines or other methods, and to reprogram the metabolism of CAR T cells to promote their effector function. This provides a new idea and direction for the treatment of solid tumors with CAR T. Creative Biolabs has launched a vaccine-boosting CAR T cell therapy solution that includes in vivo CAR-T cell function analysis following Amphi-ligand vaccination.

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In vivo efficacy experiments are the most important indicators to evaluate the therapeutic activity of drugs, and the current important models for in vivo efficacy evaluations of drugs, including cell therapy products, are mainly based on heterologous tumor models constructed on severely immunodeficient mice. Creative Biolabs's one-stop CAR-T R&D platform provides different types of animal models and various index testing services for in vivo CAR-T cell function analysis after Amphi-ligand vaccination. In addition, Creative Biolabs also has rich experience in the construction of homologous mouse tumor models, and has established a rich tumor model database, which can provide customers with a full range of in vivo efficacy evaluation services.

Amph-ligand immunization

The ability of amph-ligand immunity to expand CAR-Ts in vivo can be tested by transferring CD45.1⁺ FITC-CAR-Ts into lymphocyte-depleted homologous CD45.²⁺ recipient mice, followed by two injections of amph-FITC and adjuvant.

Method	Data	Note
Add adjuvant	Fig.1 CAR-T activity after amph-FITC and adjuvant combined vaccination.¹	Fig.1 shows a significant increase in CAR-Ts after amph-FITC vaccination and increased by combination adjuvant.

Based on state-of-the-art instrumentation, Creative Biolabs provides histological imaging services for in vivo functional analysis of your CAR-T cell therapy.

Method	Data	Note
IHC	Fig.2 IHC of LNs.¹

Confocal analysis

Confocal microscopy can perform reproducible fluorescence quantitative analysis of single or multi-labeled cells, tissue specimens and live cells, so as to quantify the antigen expression, cell structure characteristics, action and mechanism of antitumor drugs of tumor cells.

Method	Data	Note
Confocal analysis	Fig.3 Confocal imaging of LNs.¹	Confocal imaging of LNs shows that over time, amph-FITC initially aggregates in the interfollicular region but divides on CD11c⁺ dendritic cells in the T cell region.

FACS analysis

Based on advanced flow cytometry instruments, Creative Biolabs is able to perform cell proliferation assays, cell viability assays, apoptosis assays, cell surface antigen assays, intracellular antigen & cytokine assays, and more.

Method	Data	Note
Flow cytometry	Fig.4 FACS analysis of cellular biodistribution of amph-ligand.¹

CAR-T proliferation

In addition to flow cytometry, Creative Biolabs also offers proliferation assays for CAR cells to provide timely feedback on your research.

Method	Data	Note
CAR-T proliferation	Fig.5 CAR-T proliferation in LNs after amph-ligand vaccination.¹	Immunization of mice with amph-pepvIII triggers EGFRvIIICAR-T proliferation in vivo.

Expansion and cytokine analysis

Creative Biolabs's platform provides a more comprehensive assessment of your CAR-T therapy by measuring cytokine release.

Method	Data	Note
Expansion and cytokine analysis	Fig.6 Expansion and cytokine polyfunctionality of circulating CART following immunization.¹	Fig.6 shows that vaccination induces a significant increase in cell proportion and CAR-T cell versatility.

Anti-tumor efficacy evaluation

Creative Biolabs provides in vivo efficacy evaluation animals including CDX (cell line-derived xenograft) models, PDX (patient-derived xenograft) models, and Synthetic models.

Method	Data	Note
Anti-tumor efficacy evaluation	Fig.7 Anti-tumor efficacy evaluation by tumor infiltrated CART cells (Enumeration, granzyme B level, Ki67 level).¹	Amph-vax significantly increased CAR-T infiltration, and these tumor-infiltrating lymphocytes expressed higher levels of granzyme B and Ki67 than unenhanced CAR-T.

Tumor growth analysis

Creative Biolabs provides tumor growth analysis services that assess the ability of tumor cells to proliferate in animals using observations of tumor growth in animals.

Method	Data	Note
Tumor growth analysis	Fig.8 Tumor growth analysis.¹	The increased CAR-T dose of the amph vaccine eliminated tumors in most animals.

Survival analysis

Method	Data	Note
Survival analysis	Fig.9 Survival analysis.¹	CAR-T metastasis combined with amph-pepvIII immunity delays tumor growth and prolongs animal survival.

Amphiphile-ligand Vaccine Boosting CAR-T Cell Solutions

In Vitro CAR-T Cell Functional Analysis with Amphiphile-ligand Coated DC
In Vivo CAR-T Cell Functional Analysis after Amphi-ligand Vaccination

For more information about CAR-T in vivo assay, you can click here to learn more. If you have a need or question about amphi-ligand vaccination, you can also contact us directly for assistance.

Reference

Ma, Leyuan et al. "Enhanced CAR-T cell activity against solid tumors by vaccine boosting through the chimeric receptor." Science. 365,6449 (2019): 162-168.