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.
Paramyxoviruses for Tumor-targeted Immunomodulation: Design and Evaluation Ex Vivo
Characterization of Human Monocyte-derived Dendritic Cells by Imaging Flow Cytometry: A Comparison between Two Monocyte Isolation Protocols
Rapid Quantification of Mitogen-induced Blastogenesis in T Lymphocytes for Identifying Immunomodulatory Drugs
Copyright © 2024 MyJoVE Corporation. All rights reserved