Resistance of Actinobacillus pleuropneumoniae to bactericidal antibody and complement is mediated by capsular polysaccharide and blocking antibody specific for lipopolysaccharide.
Actinobacillus pleuropneumoniae is resistant to complement-mediated killing, even in the presence of specific Ab. Our studies focused on identifying the mechanism(s) responsible for this resistance. Encapsulated A. pleuropneumoniae was susceptible to killing in precolostral calf serum (PCS) but not in normal serum as a complement source in the presence of anti-capsular polysaccharide (CP) IgG. In contrast, two capsule-deficient mutants were sensitive to killing in normal serum and one was sensitive to killing in PCS alone. Electron microscopy demonstrated that A. pleuropneumoniae serotype 5a synthesized a thick, adherent CP that bound anti-CP Ab distant from the outer membrane. The CP of A. pleuropneumoniae did not prevent complement activation or the attachment of C3 to the cell surface. However, the CP did limit the amount of C9, a component of the membrane attack complex, that bound to A. pleuropneumoniae in PCS. A second mechanism of serum resistance was a result of an LPS-specific Ab present in the IgG fractions of normal swine serum, swine anti-K17 serum, and guinea pig anti-K17 LPS that blocked anti-CP IgG complement-mediated killing of A. pleuropneumoniae. Incubation of swine anti-K17 IgG with purified K17 LPS depleted Abs specific for K17 LPS but not for K17 proteins and removed all blocking activity. Immune swine serum containing this blocking Ab reduced the deposition of C9 on A. pleuropneumoniae in the presence of anti-CP IgG and also directed the deposition of C9 to sites on the bacteria in which the bound C9 was easily eluted. Thus, CP and anti-LPS Ab may act synergistically or at different stages of infection to limit the ability of complement to eliminate A. pleuropneumoniae.
Ward
C K
CK
Center for Molecular Medicine and Infectious Diseases, Virginia-Maryland Regional College of Veterinary Medicine, Virginia Polytechnic Institute, Blacksburg 24061.
Inzana
T J
TJ
D016977:Actinobacillus pleuropneumoniae / Q000276:immunology*; D000328:Adult; D000907:Antibodies, Bacterial / Q000276:immunology*; D016667:Bacterial Capsules / Q000276:immunology*; D001770:Blood Bactericidal Activity; D003167:Complement Activation; D003165:Complement System Proteins / Q000276:immunology / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D008070:Lipopolysaccharides / Q000276:immunology; D011135:Polysaccharides, Bacterial / Q000276:immunology*
D000907:Antibodies, Bacterial; D008070:Lipopolysaccharides; D011135:Polysaccharides, Bacterial; D003165:Complement System Proteins