Inhibition of NF-κB during human dendritic cell differentiation generates anergy and regulatory T-cell activity for one but not two human leukocyte antigen DR mismatches

• Published in: Human immunology 2007-09, Vol.68 (9), p.715-729
• Main Authors: Hernandez, Ana, Burger, Melissa, Blomberg, Bonnie B, Ross, William A, Gaynor, Jeffrey J, Lindner, Inna, Cirocco, Robert, Mathew, James M, Carreno, Manuel, Jin, Yidi, Lee, Kelvin P, Esquenazi, Violet, Miller, Joshua
• Format: Article
• Language: English
• Subjects: Allergy and Immunology; Allorecognition; Aspirin - pharmacology; Cell Differentiation - drug effects; Cell Differentiation - immunology; Cells, Cultured; Clonal Anergy - drug effects; Clonal Anergy - immunology; Dendritic cells; Dendritic Cells - cytology; Dendritic Cells - drug effects; FoxP3; Histocompatibility Testing; HLA-DR Antigens - analysis; HLA-DR Antigens - immunology; Humans; Lymphocyte Activation - drug effects; Lymphocyte Activation - immunology; NF-kappa B - antagonists & inhibitors; NF-κB inhibitors; Nitriles - pharmacology; Sulfones - pharmacology; T reg; T-Lymphocytes, Regulatory - drug effects; T-Lymphocytes, Regulatory - immunology
• ISSN: 0198-8859; 1879-1166
• DOI: 10.1016/j.humimm.2007.05.010

Summary We examined the in vitro inhibition of human monocyte-derived dendritic cells (DC) maturation via NF-κB blockade on T-cell allostimulation, cytokine production, and regulatory T-cell generation. DC were generated from CD14+ monocytes isolated from peripheral blood using GM-CSF and IL-4 for differentiation and TNF-α, IL-1β, and PGE2 as maturational stimuli with or without the NF-κB inhibitors, BAY 11-7082 (BAY-DC) or Aspirin (ASA-DC). Stimulator and responder cells were one versus two HLA-DR mismatched in direct versus indirect presentation assays. Both BAY-DC and ASA-DC expressed high levels of HLA-DR and CD86 but always expressed less CD40 compared with controls. Some experiments showed slightly lower levels of CD80. Both BAY- and ASA- allogeneic DC and autologous alloantigen pulsed DC were weaker stimulators of T cells (by MLR) compared with controls, and there was reduced IL-2 and IFN-γ production by T cells stimulated with BAY-DC or ASA-DC (by ELISPOT) (more marked results were always observed with ASA-treated DC). In addition, NF-κB blockade of DC maturation caused the generation of T cells with regulatory function (T regs) but only when T cells were stimulated by either allogeneic (direct presentation) or alloantigen pulsed autologous DC (indirect presentation) with one HLA-DR mismatch and not with two HLA-DR mismatches (either direct or indirect presentation). However, the T regs generated from these ASA-DC showed similar FoxP3 mRNA expression to those from nontreated DC. Extension of this study to human organ transplantation suggests potential therapies using one DR-matched NF-κB blocked DC to help generate clinical tolerance.