T Cell-based ³H-thymidine Assay

Screening cancer epitopes, particularly neoepitope antigens, has become increasingly important in drug discovery and immunotherapy in recent years. Creative Biolabs provides various T and B cell-based epitope analysis assays, including T cell-based 3H-thymidine assay, to analyze cancer pathogenic epitopes.

Cancer Epitope and T Cell-based 3H-thymidine Assay

Cancer antigen epitopes are specific molecules that target T and B cell receptors and induce complex immune responses. Cancer epitopes are typically derived from somatic mutations that cause tumorigenesis and may progress to various diseases and cancer. T cell proliferation analysis is a reliable and reproducible method to determine the presence and frequency of epitope-specific T cells. The use of a ³H-thymidine incorporation assay to investigate T cell proliferative response is a validated tool to screen and test cancer pathogenic epitopes.

Fig.1 Schematic representation of ³H-thymidine assay.¹

³H-thymidine Assay-Based Epitope Analysis Services at Creative Biolabs

Creative Biolabs has developed a robust T cell assay platform to provide reliable ³H-thymidine assay to our clients worldwide. Based on your objectives, our experts will design scientific experiments and conduct high-quality studies to support your research.

Workflow of Mouse Model-based Cancer Epitope Discovery Using ³H-thymidine Assay

Fig.2 The workflow of T cell-based ³H-thymidine assay. (Creative Biolabs)

Induce Epitope-specific T Cell Response

T cell proliferation detects antigen-responsive T cells generated from in vitro and in vivo models.

In Vitro Model

PBMCs as immune cells to mimick the in vivo environment and factors;
Dendritic cells as antigen presentation cells to induce cell immune activation.

In Vivo Model

Mouse models are immunized with antigens or peptides;
T lymphocytes are collected from mouse lymph nodes for analysis.

T Cell Proliferation Analysis

T cells cultures are incubated with ³H-thymidine for several hours to days.
As a result, dividing cells incorporated the ³H-labeled thymidine into the replicated DNA strand during the co-culturing period.
Afterward, the radioactive is measured using a scintillation counter.

Data Interpretation

The data interpretation is a necessary aspect of the assay. We have established standardized protocols to identify a cut-off value to distinguish positive results from negative ones.

Fig.3 Cell proliferation to human thyroglobulin peptides.²

Highlights of Our T Cell-based ³H-thymidine Assay

Simple and easy to perform
Relatively high throughput
Test large numbers of antigens/peptides

Creative Biolabs is a professional provider of bioassays. Our years of program development services provided us with a wealth of experience, allowing us to collaborate closely with our clients to provide T cell assays, including design and operation. Our assay provides clients with consistent and reproducible results. Please feel free to contact us us to get more information.

References

Di Blasi, D.; et al. Guidelines for low-frequency antigen-specific T cell results analysis: Dye-based proliferation assay vs 3H-thymidine incorporation. Journal of immunological methods. 2020, 487: 112907.
Flynn, J. C.; et al. Pathogenic human thyroglobulin peptides in HLA-DR3 transgenic mouse model of autoimmune thyroiditis. Cellular immunology. 2004, 229(2): 79–85.

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T Cell-based 3H-thymidine Assay