Reactivity of Haemophilus influenzae type b anti-pili antibodies

Abstract

The reactivity of anti-pilus antibodies to native and denatured Haemophilus influenzae b (Hib) pili was studied using rabbit serum prepared against piliated H. influenzae b strain M43 (p +) and adsorbed with its non-piliated variant, strain M42 (p −). The specificity of the adsorbed serum for Hib pili was documented by immunogold electron microscopy and by immunoprecipitation, which revealed the 24 kDa pilin band from strain M43 (p +) that was not seen on strain M42 (p −1). In immunodot assays, the anti-pilus antibodies reacted with the native pili present on the outer membrane of strain M43 (p +), but on Western blot assay using denatured outer membranes, the anti-pilus antibodies did not react with the 24 kDa pilin subunit. These data demonstrate that the anti-pilus antibodies in the adsorbed serum recognize conformational epitopes that depend on the tertiary or quaternary structure of Hib pili.