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Tumour infiltrating lymphocyte (TIL) and dissociated tumour cell assays

Assays for investigating the impact of therapeutics on dissociated cells from human tumours

Whilst studying therapeutic responses of human primary immune cells from blood can support the development of therapeutics at several stages of the drug discovery process, for immuno-oncology therapeutics a key milestone is demonstrating a therapeutic impact on freshly isolated cells from human tumours. Celentyx perform functional assays using dissociated cells from freshly resected human tumours. These will contain a mixture of tumour cells, stromal cells and immune cells. Importantly immune cell phenotype within the tumour may be different to blood, and so quantifying responses in this microenvironment is highly relevant to therapeutic impact. Detailed phenotyping of multiple immune cell subsets can be performed by multi-parameter flow cytometry providing a detailed understanding of their responses, which in conjunction with of multiplex analysis of supernatants by Luminex, enables a detailed characterisation of the cytokine and cytotoxic response.

Phenotyping

Analysis by spectral flow cytometry of dissociated tumour cells treated with a range of immuno-modulators.

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
T Cell Exhaustion Assays
Tumour Cell Killing Assays

Activation/exhaustion marker profiling (example below for the CD4 T cell subset):

Functional Studies

Enhancement of IFNγ production by dissociated tumour cells by Keytruda. Bars represent mean ± SEM, points represent individual donors.

Example of modulators to modify cytokine production from FOXP3+ cells (following CD4+CD127low gating) Example shown here is intracellular IFNγ. Modulators 2, 3 and 4 displaying greatest ability to enhance IFNγ production.

Target Engagement

Representative flow cytometry plot demonstrating reduction in PD-1 labelling in T cell populations by Test Drug.