Contribution of novel choline-binding proteins to adherence, colonization and immunogenicity of Streptococcus pneumoniae.
The surface of Streptococcus pneumoniae is decorated with a family of choline-binding proteins (CBPs) that are non-covalently bound to the phosphorylcholine of the teichoic acid. Two examples (PspA, a protective antigen, and LytA, the major autolysin) have been well characterized. We identified additional CPBs and characterized a new CBP, CbpA, as an adhesin and a determinant of virulence. Using choline immobilized on a solid matrix, a mixture of proteins from a pspA-deficient strain of pneumococcus was eluted in a choline-dependent fashion. Antisera to these proteins passively protected mice challenged in the peritoneum with a lethal dose of pneumococci. The predominant component of this mixture, CbpA, is a 75-kDa surface-exposed protein that reacts with human convalescent antisera. The deduced sequence from the corresponding gene showed a chimeric architecture with a unique N-terminal region and a C-terminal domain consisting of 10 repeated choline-binding domains nearly identical to PspA. A cbpA-deficient mutant showed a >50% reduction in adherence to cytokine-activated human cells and failed to bind to immobilized sialic acid or lacto-N-neotetraose, known pneumococcal ligands on eukaryotic cells. Carriage of this mutant in an animal model of nasopharyngeal colonization was reduced 100-fold. There was no difference between the parent strain and this mutant in an intraperitoneal model of sepsis. These data for CbpA extend the important functions of the CBP family to bacterial adherence and identify a pneumococcal vaccine candidate.
Rosenow
C
C
The Laboratory of Molecular Infectious Diseases, The Rockefeller University, New York, NY 10021-6399, USA.
Ryan
P
P
Weiser
J N
JN
Johnson
S
S
Fontan
P
P
Ortqvist
A
A
Masure
H R
HR
D000595:Amino Acid Sequence; D000818:Animals; D001422:Bacterial Adhesion / Q000502:physiology*; D001426:Bacterial Proteins / Q000032:analysis / Q000235:genetics; D002352:Carrier Proteins / Q000032:analysis / Q000235:genetics / Q000502:physiology*; D002460:Cell Line / Q000382:microbiology; D002794:Choline / Q000378:metabolism*; D003001:Cloning, Molecular; D015870:Gene Expression; D014644:Genetic Variation; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D010641:Phenotype; D051381:Rats; D017207:Rats, Sprague-Dawley; D017422:Sequence Analysis, DNA; D013296:Streptococcus pneumoniae / Q000737:chemistry / Q000254:growth & development* / Q000276:immunology*
D001426:Bacterial Proteins; D002352:Carrier Proteins; D002794:Choline