γδ T Cell Cytotoxicity Test

In recent years, Gamma delta (γδ) T cells have become a potential avenue for adoptive immunotherapy. To date, a number of clinical trials have used γδ T cells for cancer treatment. And this type of cell has special characteristics: MHC-unrestricted cytotoxicity, secrete inflammatory cytokines, ability to distinguish transformed cells from normal cells, as well as promote the formation of antigen-specific CD81 and CD41 T cell response. Due to the important function of γδ T cell in several diseases, it is necessary to develop suitable clinical assays for γδ T cell activity and cytotoxicity.

Creative Biolabs is a leader in the research of γδ T cells and cancer immunotherapy, we provide comprehensive γδ T cells cytotoxicity test services, including LDH cytotoxicity test, flow cytometry-based cytotoxicity test, chromium-51 release assay, impedance-based label-free real-time cytotoxicity assay…

LDH Cytotoxicity Test

The LDH-based cytotoxicity assay is one of the most commonly used methods for cell cytotoxicity detection. Lactate dehydrogenase (LDH) is a soluble cytosolic enzyme that exists in most eukaryotic cells, and when the plasma membrane is damaged upon cell death, LDH releases into the culture medium. It is a well-established and reliable indicator of cellular toxicity and can be measured by a coupled enzymatic reaction in which LDH catalyzes the conversion of lactate to pyruvate via NAD+ reduction to NADH. Then the diaphorase uses NADH to reduce a tetrazolium salt (INT) to a red formazan product that can be detected at 490 nm.

Flow Cytometry-Based Cytotoxicity Test

As an alternative to the standard chrominum-51 (51Cr) release assays, flow cytometry-based cytotoxicity test also can be used for the γδ T Cell Cytotoxicity detection. To perform the flow cytometry-based cytotoxicity assay, the γδ T Cells should be labeled with a cell tracking dye- carboxyfluorescein diacetate succinimidyl ester (CFSE) and incubated with effector cells. Then, 7-AAD, which binds to DNA and can only enter membrane-compromised cells, is used as an indicator of cell death.

Fig.1 Flow cytometric analysis of dead cell stain. (Kandarian, 2017)

Impedance-Based Label-Free Real-Time Cytotoxicity Assay

The impedance-based label-free real-time cytotoxicity assay is developed based on electrical impedance measurements and enables to achieve the real-time monitoring of T-cell-mediated cytotoxicity. This method has several advantages, such as easily feasible, sensitive and reproducible; compared to classical chromium release assay, no radioactivity is required and extended incubation periods are possible…

γδ T cells are being exploited increasingly for cancer immunotherapy. Based on our advanced technology and abundant experience, Creative Biolabs offers high-quality γδ T cells cytotoxicity test services. We also provide other customized services covering the entire γδ T cell development process to best suit your technical, program, and budget requirements. Please contact us for your exclusive solution.

Reference

1. Kandarian, F., et al. A flow cytometry-based cytotoxicity assay for the assessment of human NK cell activity. JoVE (Journal of Visualized Experiments). 2017, (126), e56191.