Soluble Antigen Arrays for Selective Desensitization of Insulin-Reactive B Cells

Abstract

Autoimmune diseases are believed to be highly dependent on loss of immune tolerance to self-antigens. Currently, no treatments have been successful clinically in inducing autoantigen-specific tolerance, including efforts to utilize antigen-specific immunotherapy (ASIT) to selectively correct the aberrant autoimmunity. Soluble antigen arrays (SAgAs) represent a novel autoantigen delivery system composed of a linear polymer, hyaluronic acid (HA), displaying multiple copies of conjugated autoantigen. We have previously reported that soluble antigen arrays displaying proteolipid peptide (SAgA_PLP) induced tolerance to this specific multiple sclerosis (MS) autoantigen. Utilizing SAgA technology, we have developed a new ASIT as a possible type 1 diabetes (T1D) therapeutic by conjugating human insulin to HA, known as soluble antigen array insulin (SAgA_Ins). Three types were synthesized, low valency _lvSAgA_Ins (2 insulins per HA), medium valency _mvSAgA_Ins (4 insulins per HA), and, high valency _hvSAgA_Ins (9 insulins per HA), to determine if valency differentially modulates the ex vivo activity of insulin-binding B cells (IBCs). Extensive biophysical characterization was performed for the SAgA molecules. SAgA_Ins molecules were successfully used to affect the biologic activity of IBCs by inducing desensitization of the B cell antigen receptors (BCR). SAgA_Ins bound specifically to insulin-reactive B cells without blocking epitopes recognized by antibodies against the Fc regions of membrane immunoglobulin or CD79 transducer components of the BCR. Preincubation of IBCs (125Tg) with SAgA_Ins, but not HA alone, rendered the IBCs refractory to restimulation. SAgA_Ins induced a decrease in BCR expression and IP3R-mediated intracellular calcium release. Surprisingly, SAgA_Ins binding to BCR on the surface of IBCs induced the observed effects at both high and low SAgA_Ins valency. Future studies aim to test the effects of SAgA_Ins on disease progression in the VH125.NOD mouse model of T1D.