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Infiltration, accumulation, and survival of chimeric antigen receptor T (CAR-T) cells in solid tumors are crucial for tumor clearance. T cell infiltration assays ensure the functional T cells are located in tumor microenvironment to display humoral or cellular immunity both in vitro and in vivo. https://www.creative-biolabs.com/car-t/enhancement-or-inhibition-of-t-cell-response-assays.htm
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Enhancement or Inhibition of T Cell Response Assays Reference: https://www.creative-biolabs.com/car-t/enhancement-or-inhibition-of-t-cell-response-assays.htm
Background • Adoptive therapy with chimeric antigen receptor (CAR) T cells is an innovative approach to program the immune system to attack malignant tumor. It uses genetically engineered immune T cells to recognize specific proteins on tumor cells through corresponding humoral immunity and cellular immunity. Except for safety, efficiency of CAR-T cell is quite crucial important to eliminate various kinds of tumors.
Proliferation Assay • T cell activation induces T cell proliferation, with the goal of clonally selecting and expanding antigen-specific T cells which can induce an effective, long-lasting adaptive immunity. To detect the extent of proliferation, we have CFSE assay and 3H-thymidine incorporation assay. • Carboxyfluorescein succinimidyl ester (CFSE) is a fluorescent cell staining dye which can be used to monitor lymphocyte proliferation, both in vitro and in vivo. The thymidine incorporation assay utilizes a strategy wherein a radioactive nucleoside, 3H-thymidine, is incorporated into new strands of chromosomal DNA during mitotic cell division. Both assays can be applied to determine the extent of T cell proliferation that has occurred in response to specific T cell stimulation.
Figure 1: T cell response - proliferation and differentiation (Butterfield, et.al. 2017)
Differentiation Assay • T cell activation induces T cell differentiation into different subsets including T helper cell (Th1, Th2, Th17 etc.), CD8+ T cell, Treg cell etc. once stimulated by specific antigens presented by APC. The differentiation is characterized by expression of various CD surface molecules. These different subsets have important implications for tumor treatment.
Figure 2: Schematic representation of various T cell subsets (O'Shea and Paul. 2010)
Functional Cytokines Producing Assay • The recognition between TCR and specific antigens presented by APC can induce T cell activation and a series of functional cytokines secretion. Cytokines are small cell-signaling protein molecules involved in cellular communication, immune response, inflammation, hematopoiesis and they are crucial for determining anti-tumor responses.
Figure 3: Different functional cytokines (Butterfield, et.al. 2017)
Cytotoxic T Lymphocyte Function Assay • Evaluation of cell-mediated immunity is significant to understand the immunotherapy. Cytotoxic T lymphocyte response is recognized as perhaps the most relevant functional measure that reflects cell-mediated acquired immune defense against cancer. Therefore, CTL assay is an excellent candidate for evaluation of the cytolytic activities.
T Cell Infiltration Assay • Infiltration, accumulation, and survival of chimeric antigen receptor T (CAR-T) cells in solid tumors are crucial for tumor clearance. • T cell infiltration assays ensure the functional T cells are located in tumor microenvironment to display humoral or cellular immunity both in vitro and in vivo.
Figure 5. CAR-T infiltrated tumor microenvironment (Newick, et.al. 2016)
Thanks Reference: https://www.creative-biolabs.com/car-t/enhancement-or-inhibition-of-t-cell-response-assays.htm
