The (alpha2-->8)-linked polysialic acid capsule and lipooligosaccharide structure both contribute to the ability of serogroup B Neisseria meningitidis to resist the bactericidal activity of normal human serum.
The molecular basis for the resistance of serogroup B Neisseria meningitidis to the bactericidal activity of normal human sera (NHS) was examined with a NHS-resistant, invasive serogroup B meningococcal isolate and genetically and structurally defined capsule-, lipooligosaccharide (LOS)-, and sialylation-altered mutants of the wild-type strain. Expression of the (alpha2-->8)-linked polysialic acid serogroup B capsule was essential for meningococcal resistance to NHS. The very NHS-sensitive phenotype of acapsular mutants (99.9 to 100% killed in 10, 25, and 50% NHS) was not rescued by complete LOS sialylation or changes in LOS structure. However, expression of the capsule was necessary but not sufficient for a fully NHS-resistant phenotype. In an encapsulated background, loss of LOS sialylation by interrupting the alpha2,3 sialyltransferase gene, lst, increased sensitivity to 50% NHS. In contrast, replacement of the lacto-N-neotetraose alpha-chain (Galbeta1-4GlcNAcbeta1-3Galbeta1-4Glc) with glucose extensions (GlcN) in a galE mutant resulted in a strain resistant to killing by 50% NHS at all time points. Encapsulated meningococci expressing a Hep2(GlcNAc)-->KDO2-->lipid A LOS without an alpha-chain demonstrated enhanced sensitivity to 50% NHS (98% killed at 30 min) mediated through the antibody-dependent classical complement pathway. Encapsulated LOS mutants expressing truncated Hep2-->KDO2-->lipid A and KDO2-->lipid A structures were also sensitive to 50% NHS (98 to 100% killed at 30 min) but, unlike the wild-type strain and mutants with larger oligosaccharide structures, they were killed by hypogammaglobulinemic sera. These data indicate that encapsulation is essential but that the LOS structure contributes to the ability of serogroup B N. meningitidis to resist the bactericidal activity of NHS.
Kahler
C M
CM
Departments of Medicine, and VA Medical Center, Atlanta, Georgia 30033, USA.
Martin
L E
LE
Shih
G C
GC
Rahman
M M
MM
Carlson
R W
RW
Stephens
D S
DS
D016667:Bacterial Capsules / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D001425:Bacterial Outer Membrane Proteins / Q000276:immunology; D001426:Bacterial Proteins / Q000235:genetics; D001770:Blood Bactericidal Activity; D002240:Carbohydrate Sequence; D005798:Genes, Bacterial; D005964:Glucosyltransferases / Q000235:genetics; D006602:Hexosyltransferases; D006801:Humans; D008070:Lipopolysaccharides / Q000096:biosynthesis* / Q000737:chemistry / Q000276:immunology; D008969:Molecular Sequence Data; D017351:N-Acetylglucosaminyltransferases / Q000235:genetics; D009345:Neisseria meningitidis / Q000145:classification / Q000235:genetics / Q000276:immunology*; D010733:Phosphoglucomutase / Q000235:genetics; D012703:Serotyping; D012794:Sialic Acids / Q000096:biosynthesis* / Q000737:chemistry / Q000276:immunology; D012799:Sialyltransferases / Q000235:genetics; D014534:UDPglucose 4-Epimerase / Q000235:genetics
D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D008070:Lipopolysaccharides; D012794:Sialic Acids; C023023:lipid-linked oligosaccharides; C021319:polysialic acid; D005964:Glucosyltransferases; D006602:Hexosyltransferases; D017351:N-Acetylglucosaminyltransferases; C103165:lgtF protein, Neisseria meningitidis; C501545:rfaK protein, Neisseria meningitidis; D012799:Sialyltransferases; D014534:UDPglucose 4-Epimerase; D010733:Phosphoglucomutase