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PubMed1999

The rgg gene of Streptococcus pyogenes NZ131 positively influences extracellular SPE B production.

Streptococcus pyogenes produces several extracellular proteins, including streptococcal erythrogenic toxin B (SPE B), also known as streptococcal pyrogenic exotoxin B and streptococcal proteinase. Several reports suggest that SPE B contributes to the virulence associated with S. pyogenes; however, little is known about its regulation. Nucleotide sequence data revealed the presence, upstream of the speB gene, of a gene, designated rgg, that was predicted to encode a polypeptide similar to previously described positive regulatory factors. The putative Rgg polypeptide of S. pyogenes NZ131 consisted of 280 amino acids and had a predicted molecular weight of 33,246. To assess the potential role of Rgg in the production of SPE B, the rgg gene was insertionally inactivated in S. pyogenes NZ131, which resulted in markedly decreased SPE B production, as determined both by immunoblotting and caseinolytic activity on agar plates. However, the production of other extracellular products, including streptolysin O, streptokinase, and DNase, was not affected. Complementation of the rgg mutant with an intact rgg gene copy in S. pyogenes NZ131 could restore SPE B production and confirmed that the rgg gene product is involved in the production of SPE B.

Amino Acid Sequence↗

PubMed1964

PURIFICATION AND PROPERTIES OF N-ACETYL-D-GLUCOSAMINE KINASE FROM STREPTOCOCCUS PYOGENES.

Zeleznick, L. D. (CIBA Pharmaceutical Co., Summit, N.J.), H. Hankin, J. J. Boltralik, H. Heymann, and S. S. Barkulis. Purification and properties of N-acetyl-d-glucosamine kinase from Streptococcus pyogenes. J. Bacteriol. 88:1288-1295. 1964.-A kinase from Streptococcus pyogenes which catalyzes adenosine triphosphate-dependent phosphorylation of d-glucose and N-acetyl-d-glucosamine has been purified 1,500-fold. The ratio of the enzymatic activity on both substrates remained constant throughout the fractionation. Similarity in heat stability, p-hydroxymercuribenzoate inhibition, protection by either carbohydrate, and lack of repression of enzymatic activity when the bacteria were grown exclusively on one of the two substrates supports the hypothesis that kinase activity is associated with one enzyme.

Acetylglucosamine↗

PubMed1989

The emergence of erythromycin resistance in Streptococcus pyogenes in Fremantle, Western Australia.

Erythromycin resistance in Streptococcus pyogenes is unusual except in Japan. Since January 1, 1985, a dramatic increase has occurred in the prevalence of erythromycin-resistant Strept. pyogenes infections in the outpatients who have presented to Fremantle Hospital, Western Australia. In 1985, 1% of isolates of Strept. pyogenes was erythromycin-resistant. This had risen to 9.1% of isolates in 1986 and to 17.6% of isolates in 1987. Several M- and T-types of Strept. pyogenes were involved. Treatment failure now can be expected when erythromycin is prescribed for the treatment of common infections in outpatients at this hospital. Laboratories that use disc methods of antimicrobial susceptibility testing may not detect erythromycin resistance in these organisms.

Adolescent↗

PubMed1981

Potentiation of opsonization and phagocytosis of Streptococcus pyogenes following growth in the presence of clindamycin.

Streptococcus pyogenes, bearing M-protein on its surface, resists opsonization by normal human serum and subsequent phagocytosis by human polymorphonuclear leukocytes. Previous studies have shown that M-protein positive organisms are poorly opsonized by the alternate pathway of complement. In an attempt to define further the role of the surface components of S. pyogenes in this process, we examined the ability of clindamycin, an antibiotic that inhibits protein biosynthesis, to alter bacterial opsonization. An M-protein positive strain of S. pyogenes was grown in varying concentrations of clindamycin at levels lower than those which inhibited growth, i.e., at levels less than the minimal inhibitory concentration. These bacteria were incubated with purified human polymorphonuclear leukocytes and peripheral blood monocytes. Significant enhancement of bacterial opsonization, phagocytosis, and killing resulted. Measurement of complement consumption and binding of the third component of complement (C3) onto the bacterial surface demonstrated that organisms grown in the presence of clindamycin activated complement more readily and fixed more C3 on their surface. Electron microscopy revealed the probable basis for these findings. Streptococci exposed to clindamycin during growth were largely denuded of surface "fuzz," the hairlike structures bearing M-protein. We conclude that the incorporation of clindamycin at concentrations that fail to inhibit growth of S. pyogenes nevertheless causes significant changes in the capacity of these bacteria to resist opsonization by serum complement. These findings support the hypothesis that M-protein inhibits bacterial opsonization by interfering with effective complement activation on the bacterial surface.

Antigens, Bacterial↗

PubMed2001

Meningitis in a girl with recurrent otitis media caused by Streptococcus pyogenes--otitis media has to be treated appropriately.

Streptococcus pyogenes rarely causes meningitis. A recent increase in the incidence and severity of diseases due to S. pyogenes has been observed worldwide, without an apparent increase in the incidence of S. pyogenes meningitis. However, more recently severe and fulminant cases of S. pyogenes meningitis have been reported in the literature. This case report emphasizes the fact that S. pyogenes can cause meningitis with severe clinical sequelae such as hygromas and right-sided third cranial nerve palsy. Most importantly, it is concluded that recurrent otitis media has to be treated carefully following appropriate identification of the causing organism in order to prevent severe clinical courses of streptococcal infections.

Adrenal Cortex Hormones↗

PubMed2001

Intra- and interspecies signaling between Streptococcus salivarius and Streptococcus pyogenes mediated by SalA and SalA1 lantibiotic peptides.

Streptococcus salivarius 20P3 produces a 22-amino-acid residue lantibiotic, designated salivaricin A (SalA), that inhibits the growth of a range of streptococci, including all strains of Streptococcus pyogenes. Lantibiotic production is associated with the sal genetic locus comprising salA, the lantibiotic structural gene; salBCTX genes encoding peptide modification and export machinery proteins; and salYKR genes encoding a putative immunity protein and two-component sensor-regulator system. Insertional inactivation of salB in S. salivarius 20P3 resulted in abrogation of SalA peptide production, of immunity to SalA, and of salA transcription. Addition of exogenous SalA peptide to salB mutant cultures induced dose-dependent expression of salA mRNA (0.2 kb), demonstrating that SalA production was normally autoregulated. Inactivation of salR encoding the response regulator of the SalKR two-component system led to reduced production of, and immunity to, SalA. The sal genetic locus was also present in S. pyogenes SF370 (M type 1), but because of a deletion across the salBCT genes, the corresponding lantibiotic peptide, designated SalA1, was not produced. However, in S. pyogenes T11 (M type 4) the sal locus gene complement was apparently complete, and active SalA1 peptide was synthesized. Exogenously added SalA1 peptide from S. pyogenes T11 induced salA1 transcription in S. pyogenes SF370 and in an isogenic S. pyogenes T11 salB mutant and salA transcription in S. salivarius 20P3 salB. Thus, SalA and SalA1 are examples of streptococcal lantibiotics whose production is autoregulated. These peptides act as intra- and interspecies signaling molecules, modulating lantibiotic production and possibly influencing streptococcal population ecology in the oral cavity.

Amino Acid Sequence↗

PubMed1991

Streptococcus pyogenes vulvovaginitis in children in Nottingham.

Isolates of Streptococcus pyogenes from vaginal swabs of children with vulvovaginitis received at Nottingham Public Health Laboratory during 1986-9 were studied. A total of 159 isolates was made during the 4 years, increasing from 17 in 1986 to 64 in 1989 and accounting for 11% of all vaginal swabs received from children. The numbers of throat swabs yielding S. pyogenes also showed an increase from 974 in 1986 to 1519 in 1989. A winter peak of isolates was noted for both vaginal swabs and throat swabs. A total of 98 strains from vaginal swabs were serotyped: 22 different types were identified, 61% of which were the common types M4, M6, R28 and M12. Erythromycin sensitivity was done on 89 strains; 84% were highly sensitive (MIC less than 0.03 mg/l). There are no other reports of such large numbers in the literature; the reason for seeing this increase in Nottingham is unclear.

Child↗

PubMed1995

Characterization of an erythromycin resistance (erm) plasmid in Streptococcus pyogenes.

Three erythromyxin-resistant Swedish isolates of Streptococcus pyogenes, representing different T-types, were studied. Two of the strains showed constitutive high-level (MIC > 200 micrograms/ml) and one showed moderate (MIC 6.4 micrograms/ml) resistance; the latter strain was sensitive to lincosamide and clindamycin, and resistance was not induced by erythromycin. In each of the strains, a plasmid with an estimated Mw of 17.6 +/- 0.9 x 10(6) was isolated in addition to smaller cryptic plasmids. The three plasmids pSE701, pSE702, and pSE703 had very similar restriction enzyme cleavage patterns. Novobiocin curing of the high-level resistance strain ER559 showed the resistance to be linked to its 17.6 x 10(6) plasmid, pSE703. Furthermore, by electroporation this rather large plasmid was reintroduced into an erythromycin-sensitive cured derivative, acquiring resistance, and the plasmid was again recovered from the transconjugant. One of the plasmids, pSE702, was shown by filter mating to be conjugative within S. pyogenes. DNA-DNA hybridization showed that the resistance determinant of the present three isolates was related to the erm gene on plasmid pAM beta 1 of Enterococcus faecalis but not to that of plasmid pE194 of Staphylococcus aureus. The copy numbers of pSE702 and pSE703, derived from the two high-level resistant strains, were 11 +/- 3 and 17 +/- 5 compared to 2 +/- 1 for pSE701, derived from the moderately resistant strain, possibly accounting for the phenotypic variation observed. The plasmids pSE702 and pAM beta 1 showed about 80% homology in DNA-DNA hybridization tests and high similarity in their restriction maps.

Drug Resistance, Microbial↗

PubMed1984

Stabilization and polysaccharide storage in group A Streptococcus pyogenes.

Water-shock treatment of group A Streptococcus pyogenes released a mixture of nucleotide-like substances and small amounts of protein. The amount of protein was much less than found with osmotic shock of Gram-negative bacteria. In group A S. pyogenes the osmolytes released exhibited as much as a 6-fold change in respect to different growth phases. Osmolyte release was dependent on the stabilization agent used and independent of cellular metabolic activity. The released osmolytes were found to be required for optimal intracellular iodophilic polysaccharide (IPS) storage. Stabilization of washing solutions, and IPS storage medium with metabolically inert non-ionic organic compounds prevented osmolyte loss and enhanced IPS storage. Polyvinyl pyrrolidone and polyethyleneglycol (MW greater than 1000) exhibited the same protective effects as found with calf serum. Smaller non-ionic organic compounds provided similar protective action but the bacteria were more susceptible to osmotic stress.

Maltose↗

PubMed1998

Survey of macrolide resistance phenotypes in Swedish clinical isolates of Streptococcus pyogenes.

Two hundred selected Swedish clinical strains of Streptococcus pyogenes, identified as erythromycin-resistant and isolated between 1980 and 1988, and 37 consecutive, resistant strains from 1989-90 were examined for resistance phenotype by disc diffusion. Strains constitutively resistant to macrolides, lincosamides and streptogramin B were absent in 1980-85 but accounted for 10% in 1986-88. The majority of the isolates belonged to a recently reported, non-inducible phenotype, described as having low-level resistance to erythromycin and sensitivity to clindamycin (82% in 1980-85, 50% in 1986-88). A significant proportion of the isolates did not agree with any known phenotype and therefore were considered as having one of three novel resistance subphenotypes. Most of the 37 strains from 1989-90 belonged to a novel subphenotype.

Anti-Bacterial Agents↗

PubMed1978

The cervical lymph nodes in Streptococcus pyogenes, group A, type 50, infection in mice.

Streptococcus pyogenes, group A, type 50, one of the few group A streptococcal types naturally occuring in mice, proved highly virulent in this species after experimental infection. Intranasal infection of 96 mice (Swiss albino, NMRI, and CBA) with this microorganism induced profound reactions in the cervical lymph nodes of 69% of the animals. Histologically, two different forms of reaction were distinguishable. In 61 mice, the lymph nodes exhibited follicular and lymphoplasmacellular hyperplasia and in 9 animals suppurative lymphadenitis was present. The ability of type 50 streptococci to persist in the pharynx of mice, and the similarity of the morphological changes induced by this organism appear to make intranasal murine group A, type 50, streptococcal infection a suitable model for human streptococcal pharyngitis.

Animals↗

PubMed2003

Clinical isolates of macrolide-resistant Streptococcus pyogenes in Central Greece.

A total of 300 Streptococcus pyogenes isolates, collected during 2001 from five hospitals in the Thessalia district (Central Greece), were examined for their resistance to macrolides. Resistance to erythromycin was detected in 58 isolates (19.3%). Of these, 68.9% were susceptible to clindamycin (M-phenotype) and carried the mefA gene. Of the remaining isolates, 18 expressed the MLS(B) phenotype: 12 and six exhibited inducible and constitutive resistance to clindamycin, respectively. All of these strains were found to be ermA(TR) positive, except for four that had the ermB gene. Of the erythromycin-resistant strains, none was found to be resistant to penicillin, tetracycline or quinupristin-dalfopristin. Molecular typing by PFGE showed the presence of a limited number of clones.

Anti-Bacterial Agents↗

PubMed1989

Sequence analysis and expression in Escherichia coli of the hyaluronidase gene of Streptococcus pyogenes bacteriophage H4489A.

The hyaluronidase gene (hylP) from Streptococcus pyogenes bacteriophage H4489A was previously cloned into Escherichia coli plasmid pUC8 as a 3.1-kilobase ThaI fragment. Southern hybridization experiments confirmed the origin of this fragment in bacteriophage H4489A before determination of the nucleotide sequence of the entire fragment. Two open reading frames (ORFs) were found, the first of which specified a 39,515-molecular-weight protein identified as the bacteriophage hyaluronidase. The second ORF encoded a 65,159-molecular-weight protein of unknown function. Putative transcription and translation control sequences for each ORF were identified by using a plasmid containing a promoterless chloramphenicol acetyltransferase gene. Controlled exclusive expression of the hylP gene via the T7 polymerase-promoter system in E. coli resulted in a 40,000-dalton protein, a result consistent with the coding capacity of the hylP gene.

Amino Acid Sequence↗

PubMed2000

Importance of inoculum size and sampling effect in rapid antigen detection for diagnosis of Streptococcus pyogenes pharyngitis.

Current recommendations suggest that negative rapid Streptococcus pyogenes antigen tests be backed up with a culture, reflecting evidence that culture may have a higher sensitivity and also that testing of a second swab may yield a different (i.e., a positive) result because of variation in sample size or distribution. If the latter is common, the sensitivities of current antigen detection tests might be improved by simply increasing the amount of sample tested. The present study assessed the effect of antigen testing of two swabs extracted together compared to independent testing of each swab extracted separately for children with clinical pharyngitis. S. pyogenes grew from one or both swabs for 198 (37%) of 537 children. The combined culture was significantly (P < 0.05) more sensitive than culture of either swab alone. Compared to combined culture, antigen testing of two swabs extracted and tested together was significantly more sensitive than two single swab extractions (94.1 versus 80%; P = 0.03); however, the specificity was decreased (81.5 versus 89.8 to 92.7%; P < 0.05). This study suggests that sample size and/or uneven sample distribution may have influenced the apparent sensitivities of prior studies that compared antigen tests to a single plate culture. A strategy, such as the one used in the present study, that increases the sample size available for antigen testing (i.e., extraction of samples from both swabs) may improve detection rates to a level that will better approximate true disease status and obviate the need for backup cultures if specificity can be improved.

Adolescent↗

PubMed1985

Cell wall preparation consisting of group A carbohydrate and peptidoglycan moieties from Streptococcus pyogenes activates murine B lymphocytes.

Cell walls from Streptococcus pyogenes strain Sv (Group A, M type 3) were lysed with M1 endo-N-acetylmuramidase, and the group A-specific carbohydrate antigen was purified by Sephadex G-100 gel filtration. The initial eluting antigen (M1gA) peak was assessed for mitogenic and polyclonal lymphocyte-activating properties in murine spleen cell cultures. Good mitogenic responses were induced over a broad dose range (1-100 micrograms) with M1gA in both BALB/c and C3H/HeJ splenic cultures. Similar mitogenic responses were induced in nude (nu/nu) and nu/+ splenic cultures, suggesting that M1gA is a B cell mitogen. The M1gA induced anti-trinitrophenyl, anti-sheep erythrocytes, and anti-horse erythrocytes polyclonal plaque-forming cell responses in splenic cell cultures. Studies with purified splenic B cells and M1gA suggest that the mitogenic responses were indeed thymic independent. These studies clearly indicate that native group A carbohydrate antigen is a B cell mitogen and polyclonal B cell activator.

Animals↗

PubMed1999

Gradient plate method to induce Streptococcus pyogenes resistance.

In recent years, increasing numbers of Streptococcus pyogenes (GAS) strains displaying resistance to macrolides have been reported in Finland, Japan, Asia and Spain. Antibiotic use has been shown to be a risk factor for infection with and carriage of drug-resistant streptococci. The aim of this study was to compare in-vitro development of resistance of streptococci to beta-lactams (penicillin, amoxycillin, cefotiam and cefuroxime) and erythromycin by serial passages in subinhibitory concentrations of antibiotics (subMICs) by gradient plate method. Three clinical strains of GAS were tested. Two were susceptible to erythromycin (MIC = 0.015 mg/L and 0.013 mg/L) and one resistant. Serial passages were performed daily by gradient plate method until a four-fold increase of the MIC was achieved. GAS variants obtained after serial passages in beta-lactams had MICs increased at least four-fold. They remained susceptible to these antibiotics. With erythromycin, final MICs reached intermediate and resistant level. Results obtained in this study with erythromycin are in good correlation with clinical studies showing that prior exposure to macrolides may help to facilitate the emergence of drug-resistant strains of streptococci.

Anti-Bacterial Agents↗

PubMed2001

Crystallization and preliminary X-ray diffraction studies of the epsilonzeta addiction system encoded by Streptococcus pyogenes plasmid pSM19035.

The proteins encoded by the Streptococcus pyogenes broad-host range and low copy-number plasmid pSM19035 form a toxin-antitoxin module that secures stable maintenance by causing the death of plasmid-free segregants. The epsilonzeta protein complex was crystallized in four different forms at pH 5.0 and pH 7.0 using the vapour-diffusion method with PEG 3350 and ethylene glycol as precipitants. Three of the crystal forms were obtained in the same droplet under identical conditions at pH 5.0. One form belongs to the enantiomorphic space groups P4(3)2(1)2 or P4(1)2(1)2. For the other two, the X-ray reflection conditions match those of space group P2(1)2(1)2(1), one representing a superlattice of the other. A crystal form growing at pH 7.0 also belongs to space group P2(1)2(1)2(1), but there is no indication of a structural relationship to the other orthorhombic forms. Initially, the crystals diffracted to 2.9 A resolution and diffracted to 1.95 A after soaking at pH 7.0. A preparation of selenomethionyl epsilonzeta protein complex yielded single crystals suitable for X-ray diffraction experiments using synchrotron sources.

Bacterial Proteins↗

PubMed2002

Chemokine production by rat macrophages stimulated with streptolysin O from Streptococcus pyogenes.

The contribution of streptolysin O (SLO) from Streptococcus pyogenes to neutrophil infiltration in inflammatory lesions was determined by production of cytokine-induced neutrophil chemoattractant (CINC)-1, -2 and -3, and macrophage inflammatory protein (MIP)-1alpha by rat macrophages stimulated with SLO in culture. Active SLO induced the production of CINCs and MIP-1alpha in dose- and time-dependent manners. These inductions were ascertained by chemokine mRNA expression in macrophages. Streptolysin S was without effect. The SLO-cholesterol complex induced the chemokine production in proportion to the residual hemolytic activity of the complex. In addition, the effects of SLO on the chemokine production were confirmed by the injection of active SLO into the preformed air pouch on the back of rats. The infiltration of neutrophils into the pouch fluid (exudate) increased steadily with a lag phase of about 2 hr. The major chemokine found in exudates was MIP-1alpha but not CINCs. In this study, it became clear that active SLO, but not the inactive one, contributed to the production of MIP-1alpha and CINCs in the conditioned medium and in exudates.

Animals↗