Multivalent Soluble Antigen Arrays Exhibit High Avidity Binding and Modulation of B Cell Receptor-Mediated Signaling to Drive Efficacy against Experimental Autoimmune Encephalomyelitis
Autor: | Martin A. Leon, Cory Berkland, Chad J. Pickens, Brittany L. Hartwell |
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Rok vydání: | 2017 |
Předmět: |
0301 basic medicine
Azides Cell signaling Encephalomyelitis Autoimmune Experimental Immunoconjugates Polymers and Plastics Injections Subcutaneous Encephalomyelitis B-cell receptor Protein Array Analysis Receptors Antigen B-Cell chemical and pharmacologic phenomena Bioengineering Autoantigens Article Cell Line Biomaterials Mice 03 medical and health sciences Antigen immune system diseases Cell Adhesion Immune Tolerance Materials Chemistry medicine Animals Humans Calcium Signaling Hyaluronic Acid Receptor Cell adhesion Autoimmune disease B-Lymphocytes Cycloaddition Reaction Chemistry Experimental autoimmune encephalomyelitis medicine.disease nervous system diseases Cell biology 030104 developmental biology Gene Expression Regulation Biochemistry Alkynes lipids (amino acids peptides and proteins) Peptides |
Zdroj: | Biomacromolecules |
ISSN: | 1526-4602 1525-7797 |
Popis: | A pressing need exists for antigen-specific immunotherapies (ASIT) that induce selective tolerance in autoimmune disease while avoiding deleterious global immunosuppression. Multivalent soluble antigen arrays (SAgA(PLP:LABL)), consisting of a hyaluronic acid (HA) linear polymer backbone co-grafted with multiple copies of autoantigen (PLP) and cell adhesion inhibitor (LABL) peptides, are designed to induce tolerance to a specific multiple sclerosis (MS) autoantigen. Previous studies established that hydrolyzable SAgA(PLP:LABL), employing a degradable linker to codeliver PLP and LABL, was therapeutic in experimental autoimmune encephalomyelitis (EAE) in vivo and exhibited antigen-specific binding with B cells, targeted the B cell receptor (BCR), and dampened BCR-mediated signaling in vitro. Our results pointed to sustained BCR engagement as the SAgA(PLP:LABL) therapeutic mechanism, so we developed a new version of the SAgA molecule using non-hydrolyzable conjugation chemistry, hypothesizing it would enhance and maintain the molecule’s action at the cell surface to improve efficacy. ‘Click SAgA’ (cSAgA(PLP:LABL)) uses hydrolytically stable covalent conjugation chemistry (Copper-catalyzed Azide-Alkyne Cycloaddition (CuAAC)) rather than a hydrolyzable oxime bond to attach PLP and LABL to HA. We explored cSAgA(PLP:LABL) B cell engagement and modulation of BCR-mediated signaling in vitro through flow cytometry binding and calcium flux signaling assays. Indeed, cSAgA(PLP:LABL) exhibited higher avidity B cell binding and greater dampening of BCR-mediated signaling than hydrolyzable SAgA(PLP:LABL). Furthermore, c SAgA(PLP:LABL) exhibited significantly enhanced in vivo efficacy compared to hydrolyzable SAgA(PLP:LABL), achieving equivalent efficacy at one quarter of the dose. These results indicate that non-hydrolyzable conjugation increased the avidity of cSAgA(PLP:LABL) to drive in vivo efficacy through modulated BCR-mediated signaling. |
Databáze: | OpenAIRE |
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