Activation of complement by mannose-binding lectin on isogenic mutants of Neisseria meningitidis serogroup B.
Mannose-binding lectin (MBL) is a serum protein that has been demonstrated to activate the classical complement pathway and to function directly as an opsonin. Although MBL deficiency is associated with a common opsonic defect and a predisposition to infection, the role of the protein in bacterial infection remains unclear. We have investigated MBL binding to Neisseria meningitidis serogroup B1940 and three isogenic mutants, and the subsequent activation of the two major isoforms of C4 (C4A and C4B) by an associated serine protease, MASP. The mutants lacked expression of the capsular polysaccharide (siaD-), the lipo-oligosaccharide (LOS) outer core that prevented LOS sialylation (cpsD-), or both capsule and LOS outer core (cps-). Using flow cytometry, it was possible to detect strong MBL binding to the cps- and cpsD- mutants over a wide range of concentrations. In contrast, minimal or no MBL binding was detected on the parent organism, with binding to siaD- only at higher MBL concentrations. C4 was activated and bound by mutants that had previously bound MBL/MASP, but there was no significant difference in the amounts of C4A and C4B bound. When sialic acid residues were removed from the parent organism by neuraminidase treatment, the binding of both MBL and C4 increased significantly. Our results suggest that MBL may bind to and activate complement on these encapsulated organisms, and the major determinants of these effects are the LOS structure and sialylation.
Jack
D L
DL
Immunobiology Unit, Institute of Child Health, London, United Kingdom.
Dodds
A W
AW
Anwar
N
N
Ison
C A
CA
Law
A
A
Frosch
M
M
Turner
M W
MW
Klein
N J
NJ
D001426:Bacterial Proteins / Q000378:metabolism / Q000502:physiology*; D002352:Carrier Proteins / Q000235:genetics* / Q000378:metabolism / Q000502:physiology*; D037222:Collectins; D003167:Complement Activation / Q000276:immunology*; D003181:Complement C4 / Q000378:metabolism; D006801:Humans; D037102:Lectins / Q000378:metabolism / Q000502:physiology*; D008070:Lipopolysaccharides / Q000378:metabolism; D016254:Mutagenesis, Insertional; D009345:Neisseria meningitidis / Q000145:classification / Q000235:genetics* / Q000276:immunology; D009439:Neuraminidase / Q000378:metabolism; D011975:Receptors, Mitogen / Q000032:analysis; D012703:Serotyping
D001426:Bacterial Proteins; D002352:Carrier Proteins; D037222:Collectins; D003181:Complement C4; D037102:Lectins; D008070:Lipopolysaccharides; D011975:Receptors, Mitogen; C023023:lipid-linked oligosaccharides; D009439:Neuraminidase