title: Toll-Like Receptor-Dependent Immune Complex Activation of B Cells and Dendritic Cells.
abstract: High titers of autoantibodies reactive with DNA/RNA molecular complexes are characteristic of autoimmune disorders such as systemic lupus erythematosus (SLE). In vitro and in vivo studies have implicated the endosomal Toll-like receptor 9 (TLR9) and Toll-like receptor 7 (TLR7) in the activation of the corresponding autoantibody producing B cells. Importantly, TLR9/TLR7-deficiency results in the inability of autoreactive B cells to proliferate in response to DNA/RNA-associated autoantigens in vitro, and in marked changes in the autoantibody repertoire of autoimmune-prone mice. Uptake of DNA/RNA-associated autoantigen immune complexes (ICs) also leads to activation of dendritic cells (DCs) through TLR9 and TLR7. The initial studies from our lab involved ICs formed by a mixture of autoantibodies and cell debris released from dying cells in culture. To better understand the nature of the mammalian ligands that can effectively activate TLR7 and TLR9, we have developed a methodology for preparing ICs containing defined DNA fragments that recapitulate the immunostimulatory activity of the previous "black box" ICs. As the endosomal TLR7 and TLR9 function optimally from intracellular acidic compartments, we developed a facile methodology to monitor the trafficking of defined DNA ICs by flow cytometry and confocal microscopy. These reagents reveal an important role for nucleic acid sequence, even when the ligand is mammalian DNA and will help illuminate the role of IC trafficking in the response.
authors.affiliation: Department of Microbiology, Boston University School of Medicine, Boston, MA, 02118, USA.
authors.forename: Krishna L
authors.initials: KL
authors.lastname: Moody
authors.affiliation: Department of Microbiology, Icahn School of Medicine at Mt Sinai, New York, NY, 10029, USA.
authors.forename: Melissa B
authors.initials: MB
authors.lastname: Uccellini
authors.affiliation: Department of Microbiology, Boston University School of Medicine, Boston, MA, 02118, USA.
authors.forename: Ana M
authors.initials: AM
authors.lastname: Avalos
authors.affiliation: Department of Medicine, University of Massachusetts Medical School, Worcester, MA, 01605, USA.
authors.forename: Ann
authors.initials: A
authors.lastname: Marshak-Rothstein
authors.affiliation: Department of Microbiology, Boston University School of Medicine, Boston, MA, 02118, USA. gviglian@bu.edu.
authors.forename: Gregory A
authors.initials: GA
authors.lastname: Viglianti
mesh_terms: D000818:Animals; D000936:Antigen-Antibody Complex / Q000276:immunology* / Q000378:metabolism; D001402:B-Lymphocytes / Q000276:immunology* / Q000378:metabolism*; D016207:Cytokines / Q000378:metabolism; D003713:Dendritic Cells / Q000276:immunology* / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D006863:Hydrogen-Ion Concentration; D008213:Lymphocyte Activation / Q000276:immunology*; D051379:Mice; D018613:Microscopy, Confocal; D021381:Protein Transport; D051193:Toll-Like Receptors / Q000378:metabolism*
chemical_list: D000936:Antigen-Antibody Complex; D016207:Cytokines; D051193:Toll-Like Receptors
keywords: AM14 transgenic BCR; Autoantibodies; B cells; Biotinylated DNA; Endogenous ligands; Flt3L-DCs; IFNα; Immune complex; Rheumatoid factor B cell; TLR7; TLR9