A novel streptococcal surface protease promotes virulence, resistance to opsonophagocytosis, and cleavage of human fibrinogen.
Group B streptococcus (GBS) is an important human pathogen. In this study, we sought to identify mechanisms that may protect GBS from host defenses in addition to its capsular polysaccharide. A gene encoding a cell-surface-associated protein (cspA) was characterized from a highly virulent type III GBS isolate, COH1. Its sequence indicated that it is a subtilisin-like extracellular serine protease homologous to streptococcal C5a peptidases and caseinases of lactic acid bacteria. The wild-type strain cleaved the alpha chain of human fibrinogen, whereas a cspA mutant, TOH121, was unable to cleave fibrinogen. We observed aggregated material when COH1 was incubated with fibrinogen but not when the mutant strain was treated similarly. This suggested that the product(s) of fibrinogen cleavage have strong adhesive properties and may be similar to fibrin. The cspA gene was present among representative clinical isolates from all nine capsular serotypes, as revealed by Southern blotting. A cspA(-) mutant was ten times less virulent in a neonatal rat sepsis model of GBS infections, as measured by LD(50) analysis. In addition, the cspA(-) mutant was significantly more sensitive than the wild-type strain to opsonophagocytic killing by human neutrophils in vitro. Taken together, the results suggest that cleavage of fibrinogen by CspA may increase the lethality of GBS infection, potentially by protecting the bacterium from opsonophagocytic killing.
Harris
Theresa O
TO
Division of Infectious Disease, Children's Hospital and Regional Medical Center, and University of Washington, Seattle, Washington, USA.
Shelver
Daniel W
DW
Bohnsack
John F
JF
Rubens
Craig E
CE
D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry / Q000502:physiology*; D015152:Blotting, Northern; D015139:Blotting, Southern; D015153:Blotting, Western; D002448:Cell Adhesion; D004926:Escherichia coli / Q000378:metabolism; D005340:Fibrinogen / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D019344:Lactic Acid / Q000378:metabolism; D008957:Models, Genetic; D009154:Mutation; D009504:Neutrophils / Q000378:metabolism; D009895:Opsonin Proteins / Q000378:metabolism*; D010587:Phagocytosis; D010641:Phenotype; D010957:Plasmids / Q000378:metabolism; D011134:Polysaccharides / Q000378:metabolism; D017434:Protein Structure, Tertiary; D051381:Rats; D012697:Serine Endopeptidases / Q000737:chemistry / Q000502:physiology*; D013296:Streptococcus pneumoniae / Q000201:enzymology* / Q000472:pathogenicity*
D001426:Bacterial Proteins; D009895:Opsonin Proteins; D011134:Polysaccharides; C062260:cold shock protein CS7.4, Bacteria; D019344:Lactic Acid; D005340:Fibrinogen; D012697:Serine Endopeptidases