In Vivo CAR-T Cell Functional Analysis Service with Peptide-DC Vaccination

DC vaccination can help activate the immune system and enhance T cell-mediated antitumor responses. Combining CAR-T cell therapy with peptide-DC vaccination may have the potential to enhance antitumor immune responses and improve treatment outcomes in cancer patients. Using state-of-the-art techniques, scientists at Creative Biolabs are committed to analyzing the activity of CAR-T cells with peptide-DC vaccination in vivo. In vivo CAR-T cell functional analysis typically involves studying the migration, expansion, persistence, and cytotoxicity of these engineered CAR-T cells. We choose the most appropriate animal model based on clients' research goals to assess the cytotoxicity and anti-tumor activity of CAR-T cells in vivo.

• In Vivo CAR-T Expansion and Activation Assay

Another approach to analyzing CAR-T cell function in vivo is through the use of flow cytometry to quantify the levels of CAR-T cells in different tissues and assess their phenotype and activation status. This technique involves isolating cells from tumor tissues or blood samples and staining them with fluorescently labeled antibodies to specific cell surface markers. We also monitor CAR-T infiltration through immunohistochemistry (IHC) staining. By using specific antibodies against the CAR construct or other markers present on the CAR-T cells, we visualize and quantify the infiltration of CAR-T cells in tumor tissues.

• In Vivo CAR-T Imaging Assay

In vivo imaging through bioluminescent imaging allows researchers to track the distribution and persistence of CAR-T cells within the body over time. This technique involves labeling the CAR-T cells with a bioluminescent reporter gene, such as luciferase, and imaging the cells.

• In vivo Anti-tumor Assessment

We assess the anti-tumor efficacy of CAR-T cells by monitoring and analysis of tumor growth inhibition or regression over time. Tumor size and growth rates are monitored regularly using imaging techniques such as bioluminescence imaging or caliper measurements. Survival analysis is also a crucial aspect of our assessment. In addition, other methods such as ELISA assays, cytokine profiling, and functional assays can be used to assess the cytotoxicity and anti-tumor activity of CAR-T cells in vivo.

• In vivo CAR-T Persistence Assay

In vivo CAR-T persistence assays allow researchers to track the presence of CAR-T cells over time and determine their ability to persist and target tumor cells. We provide several methods for in vivo CAR-T persistence assays, including bioluminescence imaging, flow cytometry analysis of peripheral blood, bone marrow, or tumor tissues, and quantitative PCR for detecting CAR-T cell genomic DNA.

We work closely with our clients to understand your specific requirements and goals, and we customize our services to meet your needs. Please contact us to learn more about our service and how it can benefit your research or experimental needs.

Support Data

Data 1: Vaccination with DCs loaded with WT1_236Y enhanced the therapeutic efficacy of CAR-T cells in a WT1-positive tumor mouse model.

Assays: In vivo anti-tumor activity assay

Detection Methods: Microcaliper

Fig.1 Tumor growth curves.¹

Data 2: DC vaccination enhanced the expansion and activation of CAR-T cells.

Assays: CAR-T expansion and activation assay

Detection Methods: Flow cytometry (Fig.2A), IHC (Fig.2B)

Fig.2 The expansion and activation status of CAR-T cells in peripheral blood (Fig.2A) and tumor tissues (Fig.2B).¹

Reference

1. Akahori, Yasushi, et al. "Antitumor activity of CAR-T cells targeting the intracellular oncoprotein WT1 can be enhanced by vaccination." Blood, The Journal of the American Society of Hematology 132.11 (2018): 1134-1145.