Effect of an Immunomodulatory Pharmacological Agent on the Interaction Between Dendritic Cells and CD4+ T Cells

Published: November 30, 2023

Abstract

Source: Wei, H., et al. Development and Functional Characterization of Murine Tolerogenic Dendritic Cells. J. Vis. Exp. (135), (2018).

This video demonstrates a method of converting dendritic cells (DCs) to tolerogenic DCs (TolDCs) using an immunomodulatory pharmacological agent. This conversion is confirmed by evaluating the TolDC function through their immunosuppressive effect on CD4+ T cells.

Protocol

1. Evaluate the Function of TolDCs In Vitro and In Vivo T-cell Syngeneic Proliferation Assay Sterilize all the surgical instruments via autoclaving and perform the experiment in a class II biological safety cabinet with appropriate safety procedures. Prepare MACS buffer using 0.5% bovine serum albumin (BSA) and 2 mM EDTA in 500 ml PBS. Sterilize the buffer by filtering through a 0.2 µm filter. NOTE: Keep t…

Divulgations

The authors have nothing to disclose.

Materials

CDDO-DFPA (RTA-408) Reata Pharmaceuticals in house synthesis Cell culture
Mouse GM-CSF Peprotech Inc 315-03 BMDC differentiation
Pan dendritic cell isolation kit Miltenyi Biotec Inc. 130-100-875 T cell proliferation assay
Pan Dendritic Cell Biotin-Antibody Cocktail Miltenyi Biotec Inc. 130-100-875 T cell proliferation assay
CD4+ T cell isolation kit Miltenyi Biotec Inc 130-104-454 T cell proliferation assay
CFSE BioLegend 423801 T cell proliferation assay
FcR Blocking Reagent Miltenyi Biotec Inc. 130-100-875 T cell proliferation assay
Anti-Biotin MicroBeads Miltenyi Biotec Inc. 130-104-454 T cell proliferation assay
1 ml syringe BD Biosciences 309626 T cell proliferation assay
BSA Sigma Aldrich Inc A2058 T cell proliferation assay
EDTA ThermoFisher Scientific 15575020 T cell proliferation assay
LS Column Miltenyi Biotec Inc. 130-042-401 T cell proliferation assay
Pre-Separation Filter Miltenyi Biotec Inc 130-095-823 T cell proliferation assay
Collagenase D Sigma Aldrich Inc. 11088858001 T cell proliferation assay
HBSS ThermoFisher Scientific 14025076 T cell proliferation assay
Ovalbumin (OVA) peptide 323–329 Sigma Aldrich Inc. O1641 T cell proliferation assay

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Citer Cet Article
Effect of an Immunomodulatory Pharmacological Agent on the Interaction Between Dendritic Cells and CD4+ T Cells. J. Vis. Exp. (Pending Publication), e21853, doi: (2023).

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