Killing of dsrA mutants of Haemophilus ducreyi by normal human serum occurs via the classical complement pathway and is initiated by immunoglobulin M binding.
Previously, we showed that serum resistance in Haemophilus ducreyi type strain 35000HP required expression of the outer membrane protein DsrA because the isogenic dsrA mutant FX517 is highly serum susceptible. In this study, we confirmed this finding by construction of additional serum-susceptible dsrA mutants in more recently isolated serum-resistant strains. We also demonstrated that killing of dsrA mutants required an intact classical complement cascade but not the alternative or mannan-binding lectin pathways. Between 5- and 10-fold more purified human immunoglobulin M (IgM) but not IgG was deposited onto dsrA mutant FX517 than onto parent strain 35000HP, consistent with IgM initiation of the classical cascade. Depletion of IgM, but not IgG, from complement-intact serum inhibited killing of FX517. As predicted from the amounts of IgM bound, more of the individual complement components were bound by FX517 than by parent strain 35000HP. Examination of the binding of negative regulators of complement as an explanation for serum resistance indicated that parent strain 35000HP bound more C4 binding protein and vitronectin than FX517 but not factor H. However, the degree and pattern of complement component binding observed suggested that IgM binding to the serum-susceptible mutant FX517 was responsible for the activation of the classical pathway and the observed killing of FX517 as opposed to binding of negative regulators of complement by the serum-resistant parent. We speculate that an undefined neo-epitope, possibly carbohydrate, is exposed in the dsrA mutant that is recognized by naturally occurring bactericidal IgM antibodies present in human sera.
Abdullah
Malikah
M
Department of Medicine, Campus Box 7031, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.
Nepluev
Igor
I
Afonina
Galyna
G
Ram
Sanjay
S
Rice
Peter
P
Cade
William
W
Elkins
Christopher
C
D001425:Bacterial Outer Membrane Proteins / Q000502:physiology*; D001770:Blood Bactericidal Activity; D015922:Complement C1q / Q000378:metabolism; D003176:Complement C3 / Q000378:metabolism; D003181:Complement C4 / Q000378:metabolism; D003171:Complement Pathway, Classical; D006191:Haemophilus ducreyi / Q000276:immunology*; D006801:Humans; D007075:Immunoglobulin M / Q000378:metabolism*
D001425:Bacterial Outer Membrane Proteins; D003176:Complement C3; D003181:Complement C4; D007075:Immunoglobulin M; D015922:Complement C1q