Engineered Secretory Immunoglobulin A provides insights on antibody-based effector mechanisms targetingClostridiodes difficile

Abstract

SUMMARYSecretory (S) Immunoglobin (Ig) A is the predominant mucosal antibody, which mediates host interactions with commensal and pathogenic microbes, includingClostridioides difficile. SIgA adopts a polymeric IgA structure that is bound by secretory component (SC). Despite significance, how SIgA supports diverse effector mechanisms is poorly characterized and SIgA-based therapies nonexistent. We engineered chimeric (c) SIgAs, in which we replaced SC domain D2 with a single domain antibody or a monomeric fluorescent protein, allowing us to investigate and enhance SIgA effector mechanisms. cSIgAs exhibited increased neutralization potency againstC. difficiletoxins, promoted bacterial clumping and cell rupture, and decreased cytotoxicity. cSIgA also allowed us to visualize and/or quantifyC. difficilemorphological changes and clumping events. Results reveal mechanisms by which SIgA combatsC. difficileinfection, demonstrate that cSIgA design can modulate these mechanisms, and demonstrate cSIgA’s adaptability to modifications that might target a broad range of antigens and effector mechanisms.