Choosing your effector preparation
Depending on what stage of testing you’re at or what you’re testing for will dictate the type of effector cells you’ll want to use, ranging from highly sensitive to biologically relevant.
Reporter cells: This allows for a highly sensitive assay system and is useful in determining whether your molecule of interest can elicit T cell activation. However, the biological relevance is low.
CD8+ cells: CD8+ T lymphocytes are the primary mediator of cytolytic activity, therefore, using these effector cells gives you a highly sensitive measure of TMC activity. However, this assay cannot capture the true biological response as CD8+ function is promoted from CD4+ cytokine release.
Pan T cell (CD3+): Using Pan T cells can measure the T cell mediated cytotoxicity response of CD4+, CD8+ and γδ T cells, accounting for the effects of other cells in the immunological response with the molecule of interest.
Expanded T cells: Clonal pools of expanded T cells can be used to generate large numbers of antigen specific T cells, whilst possessing a greater phenotypical similarity to the in vivo microenvironment than cell lines.
Peripheral Blood Mononuclear Cells (PBMCs): An effector preparation of PBMC cells will report a global cytotoxicity assay, where both TMC and ADCC (via NK cells) can be characterized. This assay setup can be used to put specific effects of the molecule of interest from more sensitive tests into a clinical context.
Healthy and disease state donors: Primary cells from healthy and disease state tissue can be used to generate data of improved clinical relevance, although they can be difficult to control and susceptible to variation which reduces sensitivity.