Effect of an Immunomodulatory Pharmacological Agent on the Interaction Between Dendritic Cells and CD4+ T Cells
Transcrição
Resuspend the cells at a 2 x 105 dendritic cells per milliliter concentration, and treat them with the appropriate experimental concentration of the triterpenoid of interest for 1 hour at 37 degrees Celsius.
During the last third of the incubation, resuspend the T cells at a 1 x 107 cells per milliliter concentration and 1 micromolar CFSE for 15 minutes at 37 degrees Celsius. Then, wash the cells with PBS and readjust the volume to a final concentration of 2 x 106 T cells per milliliter.
Co-culture 100 microliters of the treated dendritic cells with 100 microliters of the CFSE-labeled CD4 CD4-positive T cells in each well of a 96-well plate, and add 100 nanograms per milliliter of OVA peptide 322-329 to the cells, measuring the CFSE intensity of the T cells by flow cytometry after 2 to 3 days of incubation.
