Antibacterial and Protective Properties of Monoclonal Antibodies Reactive with Escherichia coli O111:B4 Lipopolysaccharide: Relation to Antibody Isotype and Complement-Fixing Activity

Abstract

In vitro and in vivo antibacterial and protective properties of murine monoclonal antibodies (MAbs) to Escherichia coli O111:B4 lipopolysaccharide (LPS) were evaluated in relation to antibody isotype and complement-fixing activity. Six O side chain-specific MAbs, including two IgMs and one of each IgG subclass, were analyzed for quantitative binding and C3 deposition on intact bacteria, complement-mediated bactericidal and opsonophagocytic activity, and protection against intraperitoneal infections in mice. Although C3 was deposited on bacteria in the presence of normal human serum (NHS) alone, LPS-specific MAbs increased C3 attachment in a dose-dependent manner. Bacterial killing occurred only in the presence of both antibody and complement NHS and required an intact alternative pathway. The efficiency of bacterial killing varied by antibody isotype (IgM > IgG2a > other IgG subclasses) and correlated with C3-fixing capacity. Opsonophagocytic activity of MAbs exhibited a similar isotype-related rank order. Likewise, IgM was more active than IgG, and IgG2a was superior to other IgG subclasses, in MAb-mediated protection against intraperitoneal infection. These data document the interdependent antibacterial and complement-fixing properties ofLPS-reactive MAbs and the degree to which both activities are determined by antibody class and isotype.