title: Antibodies against a synthetic peptide of SagA neutralize the cytolytic activity of streptolysin S from group A streptococci.
abstract: Virtually all group A streptococci (GAS) produce streptolysin S (SLS), a cytolytic toxin that is responsible for the beta-hemolysis surrounding colonies of the organisms grown on blood agar. SLS is an important virulence determinant of GAS, and recent studies have identified a nine-gene locus that is responsible for synthesis and transport of the toxin. SLS is not immunogenic; thus, no neutralizing antibodies are evoked during the course of natural infection. In the present study, we show that a synthetic peptide containing amino acid residues 10 to 30 of the putative SLS (SagA) propeptide [SLS(10-30)] coupled to keyhole limpet hemocyanin evoked antibodies in rabbits that completely neutralized the hemolytic activity of the toxin in vitro. Inhibition of hemolysis was reversed by preincubation of the immune serum with soluble, unconjugated peptide, indicating the specificity of the antibodies. In addition, antibodies that were affinity purified over an SLS(10-30) peptide column completely inhibited SLS-mediated hemolysis. The SLS(10-30) antisera did not opsonize group A streptococci; however, when combined with type-specific M protein antisera, the SLS antibodies significantly enhanced phagocytosis mediated by M protein antibodies. Thus, we have shown for the first time that it is possible to raise neutralizing antibodies against one of the most potent bacterial cytolytic toxins known. Our data also provide convincing evidence that the sagA gene actually encodes the SLS peptide of GAS. The synthetic peptide may prove to be an important component of vaccines designed to prevent GAS infections.
authors.affiliation: Department of Veterans Affairs Medical Center and Department of Medicine, University of Tennessee Health Science Center, Memphis, Tennessee 38104, USA. james.dale@med.va.gov
authors.forename: James B
authors.initials: JB
authors.lastname: Dale
authors.forename: Edna Y
authors.initials: EY
authors.lastname: Chiang
authors.forename: David L
authors.initials: DL
authors.lastname: Hasty
authors.forename: Harry S
authors.initials: HS
authors.lastname: Courtney
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology*; D000918:Antibody Specificity; D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins / Q000276:immunology; D001426:Bacterial Proteins; D002352:Carrier Proteins / Q000276:immunology; D007106:Immune Sera / Q000276:immunology; D008969:Molecular Sequence Data; D010587:Phagocytosis; D011817:Rabbits; D022541:Streptococcal Vaccines / Q000276:immunology; D013297:Streptococcus pyogenes / Q000276:immunology*; D013301:Streptolysins / Q000737:chemistry / Q000276:immunology*
chemical_list: D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D002352:Carrier Proteins; D007106:Immune Sera; D022541:Streptococcal Vaccines; D013301:Streptolysins; C014689:streptococcal M protein; C026050:streptolysin S