Home > Immunology Assays > T Cell Exhaustion Assays
Assays for measuring reversal of T cell exhaustion by checkpoint inhibitors, cytokines and immuno-modulatory drugsT cell exhaustion - the reduction in T cell responsiveness following chronic exposure to antigen - impairs the immune system's ability to fight cancer and infectious diseases. As part of Celentyx’s CRO services for immuno-oncology, our high-throughput assay platform using human ‘exhausted’ T cells reveals the potential of immunotherapies to reverse CD4 and CD8 T cell exhaustion.
In brief, the isolated T cells undergo an initial prolonged stimulation that promotes a state of unresponsiveness with subsequent restimulation in the absence or presence of test molecules quantifies their ability to reverse the exhausted multi-parameter phenotype including cytokine production, proliferation, activation marker expression and tumour cell killing capacity. This may enable the investigation of molecules that may block or modulate inhibitory surface receptors or their ligands (e.g. PD-1, PDL1, PDL2, CTLA-4, LAG3, TIM3, Galectin 9, TIGIT, CD155, BTLA, KLRG1, CD160, 2B4, A2aR, IL-10R, CSF1R, TGFβR), inhibitory kinases/phosphatases (e.g. SHP1, SHP2, PTPN22, PKC, PTP1B, PTPN2), agonise stimulatory receptors (e.g. CD28, CD137/4-1BB, GITR, CD40, ICOS), inhibit enzymes generating soluble suppressive mediators or involved in inhibitory pathways (e.g. CD39, CD73, IDO, TDO). The platform has proven ability to identify and quantify the potential efficacy of a wide range of modulators with a diverse array of molecular targets in screening, efficacy and benchmarking assays, thereby accelerating the drug discovery process in the field of immuno-oncology. |
Further Immunology Assays
B Cells Haemolysis Testing Human Microglia Macrophages/Monocytes Neutrophils/Granulocytes Phagocytosis Assays Regulatory T cells Spheroid Killing Assays Suppression Assays T Cell Activation Assays Tumour infiltrating lymphocyte (TIL) and dissociated tumour cell assays Tumour Cell Killing Assays |
Phenotype of exhausted T cells
Compared to unstimulated T cells, exhausted T cells exhibit up-regulation of exhaustion markers such as PD-1 and Tim-3 (A), as well as the transcription factor TOX. Upon restimulation, upregulation of activation markers (B) and cytokine production (e.g. IFNγ; C).
Reversal of T cell exhaustion by test molecules
Test molecules may reverse different facets of T cell exhaustion. A Shows benchmarking of a test molecule to the PD-1 blocking antibody, Keytruda, which restores the IFNγ response in a concentration-specific manner. B Shows the restoration of the proliferative response of exhausted cells by a test molecule.
