Steric hindrance of antibody binding to surface proteins of Coxiella burnetii by phase I lipopolysaccharide

Abstract

The exposure of surface protein antigens on virulent phase I Coxiella burnetii was compared with that on avirulent phase II cells. Although anti-phase II antibodies did not bind to the surfaces of native intace phase I cells, they bound to phase I proteins if the proteins were solubilized for sodium dodecyl sulfate-polyacrylamide gel electrophoresis and analyzed by immunoblotting. In addition, removal of the phase I lipopolysaccharide (LPS) by trichloroacetic acid exposed surface proteins for reactivity with anti-phase II antibodies, as shown by immunofluorescence assays, direct antibody binding, and immunoelectron microscopy using protein A-colloidal gold conjugates. Based on these observations, a simple model of phase variation is proposed to explain the apparently conflicting notions of the identity of the phase II antigen(s). The model suggests that the phase I LPS sterically hinders access of anti-phase II antibodies to a multitude of shared protein antigens, any one of which may confirer phase II specificity. Exposure of these shared protein antigens through the appearance of a more truncated LPS (phase II) or extraction of the smooth-type phase LPS allows antibody accessibility and confers apparent phase II serospecificity.