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The isolated monocytes are differentiated in vitro to an immature DC phenotype. These cells are then loaded with the test protein before inducing further differentiation to a mature DC phenotype.
Once matured, the cells are incubated with autologous CD4+ T cells before measuring T cell proliferation using EdU incorporation and T cell activation by assessing CD25/OX-40 upregulation.
Pre-screen tests using a small numbers of donors can be performed to evaluate if there is direct modulation of T cell activation prior to choosing a time course or DC:T cell assay.