Virulence of Staphylococcus aureus mutants altered in type 5 capsule production.
Most clinical isolates of Staphylococcus aureus produce microcapsules (uronic acid-containing extracellular polysaccharides) that are detectable by serologic methods but are not visible by negative staining. Among the 11 reported serotypes, capsule types 5 and 8 comprise approximately 75% of all isolates. Transposon mutagenesis was performed on S. aureus to create mutants altered in capsule expression. Tn918 was introduced into the capsule type 5 strain Reynolds by filter mating, and a capsule-deficient transconjugate, JL236, was isolated. The wild-type strain was transformed with JL236 chromosomal DNA to confirm that transfer of the appropriate-size chromosomal fragment containing Tn918 generated a capsule-deficient transformant. Strain Reynolds was mutagenized with ethyl methanesulfonate to obtain a capsule-negative mutant (strain JL240). Capsular phenotypes were determined by colony immunoblots, antibody adsorption experiments, and transmission electron microscopy. The virulences of the parental and mutant strains in mice were compared. The 50% lethal doses for strains Reynolds, JL236, and JL240 were similar (10(8.59), 10(8.98), and 10(8.93) CFU, respectively). Animals injected intraperitoneally with either wild-type or mutant strains had comparable levels of bacteremia at 3 and 24 h after challenge. Quantitative cultures of blood and kidneys from animals challenged intravenously with sublethal doses of the S. aureus strains also showed no differences in bacterial clearance or renal abscess formation. These studies indicate that the type 5 S. aureus microcapsule does not promote bacterial virulence in the animal models tested.
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Channing Laboratory, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115.
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D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology; D015139:Blotting, Southern; D004251:DNA Transposable Elements / Q000235:genetics; D005260:Female; D007668:Kidney / Q000382:microbiology / Q000473:pathology; D051379:Mice; D008810:Mice, Inbred C57BL; D016254:Mutagenesis, Insertional / Q000235:genetics; D011135:Polysaccharides, Bacterial / Q000235:genetics* / Q000276:immunology; D011817:Rabbits; D013203:Staphylococcal Infections / Q000382:microbiology / Q000401:mortality / Q000473:pathology; D013211:Staphylococcus aureus / Q000235:genetics / Q000276:immunology / Q000472:pathogenicity* / Q000648:ultrastructure; D014169:Transformation, Bacterial; D014774:Virulence / Q000235:genetics
D000907:Antibodies, Bacterial; D004251:DNA Transposable Elements; D011135:Polysaccharides, Bacterial