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PubMed1979

Immunofluorescent and immunoelectron microscopic demonstration of primate heart muscle antigens cross-reactive with Streptococcus pyogenes.

The presence of antigens in heart muscles cross-reactive with antibodies against Streptococcus pyogenes were examined in heart samples of seven primate species (chimpanzee, rhesus macaque, stump-tailed macaque, hamadryas baboon, capuchin monkey, green monkey and patas monkey). Cross-reactive antigens were localized by means of the indirect immunofluorescent technique in nearly all samples tested. The fluorescence pattern was the same as found in the heart muscle of other mammals. Using the indirect immunoferritin technique the substructural distribution of the antigens was investigated in heart muscle of rhesus macaque and hamadryas baboon. Beside the location on the cell surface the antigens were also demonstrated within the cell. Above all there was a strong labelling along the intercalated discs and in the sarcoplasmatic reticulum.

Animals↗

PubMed1978

Transfer of erythromycin resistance from clinically isolated lysogenic strains of Streptococcus pyogenes via their endogenous phage.

Endogenous bacteriophage from clinically isolated, erythromycin-resistant strains of Streptococcus pyogenes, upon induction by treatment with mitomycin C, were demonstrated to cause transduction of antibiotic resistance yielding transductants resistant to relatively high concentrations of erythromycin. Transduction of resistance to erythromycin via endogenous phage may play a significant role in the emergence of streptococci with multiple resistance in the clinical environment.

Bacteriophages↗

PubMed2025

The Streptococcus pyogenes mannose phosphotransferase system (Man-PTS) influences antimicrobial activity and niche-specific nasopharyngeal infection.

Streptococcus pyogenes is a human-adapted pathogen that can cause multiple diseases, including pharyngitis and skin infections. Although this bacterium produces many virulence factors, how S. pyogenes competes with the host microbiota is not well understood. Here, we detected antimicrobial activity from S. pyogenes MGAS8232 that prevented the growth of Micrococcus luteus. This activity was produced when cells were grown in 5% CO2 in M17 media supplemented with galactose; however, the addition of alternative sugars coupled with genome sequencing experiments revealed that the antimicrobial phenotype was not related to classical bacteriocins. To further determine genes involved in the production of this activity, a transposon mutant library in S. pyogenes MGAS8232 identified the mannose phosphotransferase system (Man-PTS), a major sugar transporter, as important for the antimicrobial phenotype. Loss-of-function transposon mutants linked to the antimicrobial activity were identified to also be involved in alternative sugar utilization, and additionally, the Man-PTS was further identified from an inadvertent secondary mutation in a bacteriocin operon mutant. Sugar utilization in the Man-PTS mutants demonstrated that galactose, mannose, and N-acetylglucosamine utilization was impaired. RNA-seq experiments in high and low glucose concentrations further characterized the Man-PTS as a glucose transporter; however, transcriptional regulators or virulence factors were not affected with the loss of the Man-PTS. Deletion of Man-PTS demonstrated defects in a mouse model of nasopharyngeal infection but not skin infection. This work suggests that the ability of S. pyogenes to utilize alternative sugars presented by glycans may play a role in acute infection and interactions with the endogenous microbial population existing in the nasopharynx.IMPORTANCEStreptococcus pyogenes is responsible for over 500,000 deaths per year primarily due to invasive infections and post-infection sequelae, although the most common manifestations include pharyngitis and impetigo. S. pyogenes can adapt to its environment through alternative sugar metabolism. Here, we identified an antimicrobial phenotype that was not bacteriocin-related but a by-product of alternative sugar metabolism. The mannose phosphotransferase system was involved in the production of the antimicrobial and was also important for S. pyogenes to utilize alternative sugars and establish nasopharyngeal infection but not skin infection. Overall, this study identified potential strategies used by S. pyogenes for interactions with the endogenous microbiota and further elucidated the importance of sugar metabolism in acute upper respiratory tract infection.

Streptococcus pyogenes↗

PubMed1978

[Clinical and immunological studies with OK-432 (Streptococcus pyogenes) on immunotherapy in cancer patients].

In a phase I study, the bacterial vaccine OK-432 (streptococcus pyogenes) was investigated in eight patients with advanced malignant tumors. Besides a fever reaction after i.v. application no toxic side effects were observed. Additionally, its therapeutic effectiveness could be demonstrated by i.v. and local therapy. There was also observed an increase of lymphocyte blastogenesis response in patients under treatment with OK-432.

Bacterial Vaccines↗

PubMed2025

Invasive Streptococcus dysgalactiae subspecies equisimilis compared with Streptococcus pyogenes in Australia, 2011-23, and the emergence of a multi-continent stG62647 lineage: a retrospective clinical and genomic epidemiology study.

BACKGROUND: Streptococcus dysgalactiae subspecies equisimilis (SDSE) is closely related to Streptococcus pyogenes, with overlapping disease manifestations. We compared the clinical and genomic epidemiology of invasive SDSE with invasive S pyogenes across different settings in Australia and phylogenetically contextualised the SDSE sequences within a global cohort of genomes. METHODS: In this retrospective clinical and genomic epidemiology study, cases of invasive SDSE isolated from normally sterile sites were identified and whole-genome sequenced across five hospital networks in temperate southeast Australia (Melbourne and Sydney) and the tropical Top End of the Northern Territory. SDSE disease incidence, case demographics, clinical outcomes, and longitudinal lineage dynamics were compared between southeast Australia and the Top End and to co-collected invasive S pyogenes cases in each region. SDSE genomes and lineages were also contextualised within 1166 global SDSE sequences. Genomic transmission clusters (not necessarily direct transmission) were inferred between isolates from different individuals by single-linkage clustering at a single nucleotide polymorphism threshold of less than or equal to seven for SDSE and less than or equal to five for S pyogenes based on previous transmission analyses. FINDINGS: Between Jan 1, 2011, and Feb 28, 2023, there were 693 invasive SDSE cases and 995 invasive S pyogenes cases. Invasive SDSE occurred almost exclusively in adults. The overall invasive SDSE incidence in southeast Australia was similar to invasive S pyogenes (incidence rate ratio [IRR] 1·15, 95% CI 0·91-1·46; p=0·26) and increased over the study period (IRR 1·06 per year, 95% CI 1·05-1·08; p<0·0001) from 1·30 cases per 10 000 admissions in 2011 to 3·72 cases per 10 000 admissions in the first 2 months of 2023 (95% CI 2·13-6·07). In southeast Australia, where stringent COVID-19 non-pharmaceutical interventions (NPIs) were implemented between 2020 and 2021, the SDSE incidence plateaued during 2020-21 but did not significantly decline (IRR 1·09 compared with 2017-19, 95% CI 0·88-1·35; p=0·47). By contrast, S pyogenes incidence substantially declined in 2020-21 in southeast Australia (IRR 0·35 compared to 2017-19, 95% CI 0·22-0·52; p=0·017). In the Top End, SDSE incidence was lower than S pyogenes (IRR 0·24, 95% CI 0·19-0·31; p<0·0001). However, crude incidence remained higher than southeast Australia (crude IRR 1·24, 95% CI 1·07-1·42; p=0·0037) and disproportionately affected First Nations Australians in the Top End compared with non-First Nations individuals (IRR 3·36, 95% CI 2·33-4·85; p<0·0001). Comparing 2020-21 with 2017-19, there was no decline in SDSE (IRR 1·27, 95% CI 0·73-2·24; p=0·45) or S pyogenes (IRR 0·97, 95% CI 0·80-1·18; p=0·81) incidence in the Top End, which did not implement prolonged stringent COVID-19 NPIs. Analysing the available genomes of invasive cases and in lineages for which more than or equal to five invasive cases occurred, only 24 (6%) of 384 SDSE cases were assigned to genomic transmission clusters, compared with 271 (52%) of 524 S pyogenes cases. An stG62647 lineage encompassed 113 (26%) of 436 sequenced SDSE genomes. Analysis of available SDSE sequences from Australia, western Europe, and North America inferred concurrent international expansion of the stG62647 lineage in all three regions between 1990 and 2005. INTERPRETATION: We identified a substantial burden of invasive SDSE, dominated by the emergent stG62647 lineage. The contrasting epidemiology between species in the different Australian regions, during COVID-19 NPIs, and genomic infection patterns indicates transmission dynamic, pathogen population, and host-pathogen interaction differences between SDSE and S pyogenes and indicates implications for disease control measures. FUNDING: Australian National Health and Medical Research Council.

Humans↗

PubMed2002

Influence of recombination and niche separation on the population genetic structure of the pathogen Streptococcus pyogenes.

The throat and skin of the human host are the principal reservoirs for the bacterial pathogen Streptococcus pyogenes. The emm locus encodes structurally heterogeneous surface fibrils that play numerous roles in virulence, depending on the strain. Isolates harboring the emm pattern A-C marker exhibit a strong tendency to cause throat infection, whereas emm pattern D strains are usually recovered from impetigo lesions; as a group, emm pattern E organisms fail to display obvious tissue tropisms. The peak incidence for streptococcal pharyngitis and impetigo varies with season and locale, leading to wide spatial and temporal distances between throat and skin strains. To assess any impact of niche separation on genetic variation, the extent of recombinational exchange between emm pattern A-C, D, and E subpopulations was evaluated. Analysis of nucleotide sequence data for internal portions of seven housekeeping loci from 212 isolates provides evidence of extensive recombination between strains belonging to different emm pattern subpopulations. Furthermore, no fixed nucleotide differences were found between emm pattern A-C and D strains. Thus, despite some niche separation created by distinct epidemiological trends and innate tissue tropisms there is little evidence for neutral gene divergence between throat and skin strains. Maintenance of a relationship between emm pattern and tissue tropism in the face of underlying recombination suggests that tissue tropism is associated with emm or a closely linked gene.

Alleles↗

PubMed1978

L-forms of Pseudomonas aeruginosa. 3. The serological cross-reactions among stable L-forms of Pseudomonas aeruginosa, L-form of Streptococcus pyogenes and mycoplasmas.

The serological cross-reactions among stable L-forms of Pseudomonas aeruginosa, or of Streptococcus pyogenes and mycoplasmas were investigated in passive hemagglutination (PHA) and agar gel diffusion tests. (1) PHA active fraction which reacts most strongly with homologous serum was obtained from the sodium dodecyl sulfate soluble fraction of L-form membrane after disruption by Braun cell homogenizer. (2) Antiserum prepared against L-forms in rabbits showed high-level stimulation of PHA antibodies, while it showed little stimulation of agglutinating antibodies. (3) In PHA and agar gel diffusion tests, antiserum to L-form of P. aeruginosa strain IFO-3,455 reacted with L-form of strain N-10, which is serologically different in serotype from strain IFO-3,455. Furthermore, the two L-forms of P. aeruginosa cross-reacted with a L-form of S. pyogenes. 4) L-forms of P. aeruginosa and S. pyogenes did not react with five strains of mycoplasmas in PHA and agar gel diffusion tests.

Antigens, Bacterial↗

PubMed1978

The characteristics of M proteins purified by column chromatography with hydroxyapatite from acid extracts of Streptococcus pyogenes of types 1, 3, 6, 12 and 17.

Purified M proteins were recovered from acid extracts of Streptococcus pyogenes, M-types 1, 3, 6, 12 and 17, by elution from columns of hydroxyapatite of the proteins precipitated with ammonium sulphate. M protein free from non-type-specific antigens was recovered only from M-type 12. Although similar fractions were not recovered from M-types 1, 3, 6 and 17, purified preparations containing a single cross-reactive antigen were obtained. In addition to the M proteins associated with cross-reactive antigens, type-specific antigens that did not stimulate opsonic antibodies were isolated from revealed molecular weights that ranged from 32,000 to 63,000 daltons, total amino acid compositions that were similar, and N-terminal amino acids that were variable.

Amino Acid Sequence↗

PubMed1978

Regulation of the synthesis of M protein by sugars, Todd Hewitt broth, and horse serum, in growing cells of Streptococcus pyogenes.

Various sugars were tested for their effect on the differential rate of synthesis of M protein during the growth of Streptococcus pyogenes strain 0055 M12T12. In a semisynthetic medium alone, a high rate of M protein synthesis occurred with glucose as a substrate; decreasing rates of synthesis occurred with sucrose and trehalose, in that order, although the rates of growth were approximately equal with all sugars. A period of derepressed synthesis of M protein occurred in the lag phase of growth and in the stationary period as the substrates were being depleted. Although glucose inhibited the utilization of other sugars, diauxie was not apparent from the growth curves. However, synthesis of M protein followed strong diauxie curves with a reduction in rate of synthesis during the utilization of the second sugar. With glucose as a substrate, 2-deoxyglucose showed a strong permanent repression of M protein synthesis, whereas both glucose and 2-deoxyglucose caused temporary repression when sucrose was the substrate. Horse serum increased the rate of synthesis of M protein in a manner very similar to that caused by adding cyclic AMP, although quantitative analyses suggested that cyclic AMP, per se, was not the effector in horse serum. Addition of Todd Hewitt broth permitted the organisms to grow on phosphorylated sugars. Although the rates of growth on phosphorylated sugars were similar to that obtained with glucose, M protein was not synthesized when a phosphorylated sugar was the sole substrate. The addition of phosphorylated sugars with glucose or sucrose as substrates strongly repressed the synthesis of M protein with glucose-1-phosphate and with fructose 1,6-diphosphate repressing M protein synthesis the most. Clearly, M protein synthesis, which was not required for growth, was preferentially induced by glucose as compared to the other sugars and was dependent upon the metabolic route by which glucose was utilized.

Animals↗

PubMed1976

Adaptation of an osmotically fragile L-form of Streptococcus pyogenes to physiological osmotic conditions and its ability to destroy human heart cells in tissue culture.

An osmotically fragile L-form of Streptococcus pyogenes, type 12, was quickly rendered osmotically stable by decreasing the sodium chloride content of the growth medium and with the temporary use of oleic acid. The change from osmotic fragility to stability was accompanied by changes in cell yield, generation time, saturated/unsaturated fatty acid ratio of the membrane, and cytoplasmic protein composition. Finally, this resulting osmotically stable L-form survived and was capable of rapidly destroying Girardi human heart cells in tissue culture.

Cell Survival↗

PubMed1976

[The immunobiology of M-proteins of Streptococcus pyogenes. III. Immunization of rhesus monkeys (Macaca mulatta)].

Rhesus monkeys were used for the examination of immunogenicity of various preparations of M-antigens of Streptococcus pyogenes, types 1, 12, and 19. The antigens were applied in various dosages, but always with alumniumhydroxide as adjuvant. The challenge infection was provided by the homologous type, in some cases by an additional heterologous type. After intravenous infection all the animals fell ill, as the high dose of infective agents led to poisoning. The majority of immunized animals, however, did not show an increase of antistreptolysin titers with the type 1-infections - a sign of the absence of any reproduction of the injected streptococci in the animal organism. This behaviour was also observed in animals where no protective antibodies (bactericidal and long-chain antibodies) were verified before the infection. The electrocardiographic findings also indicate the protective effect of M-protein vaccines. In all control animals changes were seen which indicated an acute myocarditis. Normal electrocardiograms were obtained from the monkeys which had been immunized and afterwards infected. The administration of antigen every two weeks over a period of one year did not lead to the occurrence of any visible allergy in the skin test. The lymphocyte transformation test showed an increased rate of transformation in some immunized and infected animals, but this is not type-specific.

Animals↗

PubMed1976

Membrane lipoteichoic acid of Streptococcus pyogenes and its stabilized L-form and the effect of two antibiotics upon its cellular content.

Membrane lipoteichoic acid continues to be synthesized by an osmotically fragile, stabilized L-form of Streptococcus pyogenes. Chromatographic and electrophoretic comparisons indicate that the lipid componenent of lipoteichoic acid in this L-form and its parental streptococcus is glycerophosphoryldiglucosyl diglyceride and not phosphatidylkojibiosyl diglyceride. Based upon dry weight determinations, the yield of lipoteichoic acid from the L-form is 0.19%, as compared with 0.97% from the streptococcus. When grown with bacitracin the L-form contains the same amount of teichoic acid as when grown without this antibiotic; however, its lipoteichoic acid content is reduced by 85%. Similarly, the L-form grown with novobiocin for 10 h contains only 17% of the teichoic acid found in control cells.

Bacitracin↗

PubMed1976

Streptococcus pyogenes preparation OK-432: immunoprophylactic and immunotherapeutic effects on the incidence of spontaneous leukemia in AKR mice.

An inactivated and lyophilized preparation of a low virulence strain (Su) of Streptococcus pyogenes (group A) was designated OK-432. When 2- and 5-month-old AKR mice were inoculated im with OK-432 twice weekly throughout their life-spans, spontaneous leukemias occurred later and at a lower incidence than in control groups. By virus neutralization and cytotoxicity tests and by immunoelectron microscopy, antibodies against virus and cell-surface antigens of transplanted AKR leukemia were not detectable in sera of nonleukemic mice of any group. Whereas sera from mice treated with OK-432 were the only positive for interferon, viremia was clearly demonstrated in control groups by reverse transcriptase assays of the plasma.

Animals↗

PubMed2026

Understanding carriage of Streptococcus pyogenes directly from the human host.

Using genomic approaches, Faiola et al.1 evaluated the Streptococcus pyogenes carriage state, and pre- and post-treatment pharyngitis, directly from human subjects. This work suggests that carriage isolates may evolve with host-specific mechanisms rather than a unified regulatory phenotype.

Journal Article↗

PubMed1977

Synthesis of "group polysaccharide" by membranes from Streptococcus pyogenes and its stabilized L-form.

Rhamnose and N-acetylglucosamine (GlcNAc) are incorporated from thymidine 5'-diphosphorhamnose and uridine 5-diphospho-N-acetylglucosamine (UDPGlcNAc) into membrane fragments prepared from Streptococcus pyogenes but not into membrane fragements prepared from a stabilized L-form of this organism. Incorporation from TDPrhamnose is partially dependent upon UDPGlcNAc and vice versa. The oligomeric GlcNAc and rhamonose-containing products are easily extracted from membrane particles by sedimentation through detergent solutions. They are substantially extracted into methanol but not into chloroform-methanol (2:1). When product containing both radioactive rhamnose and GlcNAc is deacetylated and hydrolyzed briefy in acid, glucosaminyl rhamnose is obtained, byt not higher oligomers, suggesting that oligomer synthesis in vitro is terminated because unidentified wnzymatic requirements are not satisfied. The data are consistent with the assembly of group A-specific polysaccharide at the cellular membrane with participation of a lipoid anchor (acceptor) molecule.

Acetylglucosamine↗

PubMed1976

Persistence, pathogenesis, and morphology of an L-form of Streptococcus pyogenes adapted to physiological isotonic conditions when in immunosuppressed mice.

Evidence obtained using nonimmunosuppressed and newborn mice suggests that the immune response of the host plays a role in the rapid removal of a physiologically isotonic L-form of Streptococcus pyogenes, since its inability to persist in vivo was not due to osmotic lysis. With mice immunosuppressed with methylprednisolone sodium succinate, viability and detection of this L-form by fluorescent antibody was prolonged for at least 2 weeks, the approximate duration of immunosuppression in these mice. However, heat-killed L-form cells only persisted for 3 days in such mice. Therefore, persistence of a viable L-form in these treated mice was not simply due to the lack of removal of L-forms by a compromised host. At no time was there any indication of illness in nonimmunosuppressed or immunosuppressed mice after L-form injection, and all internal organs, when examined macroscopically, remained normal. Thus, overt pathogenesis was not a characteristic of this L-form in a suitable host even when its immune response had been compromised. The microscopic morphology of the L-form after isolation from immunosuppressed mice changed drastically. It was typically micrococcal in appearance and exemplified the cellular variability achievable by this organism in vivo. Also, streptolysin S production was increased markedly by passage of the L-form in immunosuppressed mice. However, M protein, as a cellular component, was not detected serologically, nor was any reformation of a rigid cell wall apparent by electron microscopy after isolation of this streptococcal L-form from such mice.

Animals↗

PubMed1976

Membrane studies of Streptococcus pyogenes and its L-form growing in hypertonic and physiologically isotonic media. An electron spin resonance spectroscopy approach.

Electron spin resonance spectroscopy (ESR) was used to compare the lipid organization, thermal stability and the physical state of the membrane of a human pathogen, Streptococcus pyogenes and its osmotically fragile L-form with this same L-form now adapted to grow under physiologically isotonic conditions (physiological L-form). Comparison of the hyperfine splittings of a derivative of 5-ketostearic acid spin label, I(12, 3), after incorporation into the membrane, revealed that the lipid chain rigidity of these membranes is in the order physiological L-form greater than osmotically fragile L-form greater than streptococcus. The signal intensity (of the center magnetic field line) versus temperature analysis showed two transitions for these membranes. The first with melting points of 45, 26 and 36 degrees C and second transition at 70, 63 and 60 degrees C for the physiological L-form, osmotically fragile L-form and streptococcal membranes, respectively. This same order of membrane lipid chain rigidity was seen from the cooperativities obtained for each of these systems from analysis based on the expression for an n-order reaction. The I(12, 3) and other probes with the paramagnetic group close to the methyl end of the molecule suggested that this difference in lipid chain rigidity between these organisms resides in the environment closer to the lipid head group region rather than in the hydrophobic lipid core. Another major finding was the binding of I(12, 3) at two or more different sites in each of the membranes examined. This change in lipid chain rigidity now provides an explanation to account for the survival of a previously osmotically fragile L-form in physiologically isotonic media by focusing on changes in the physical nature of its membrane. In so doing, it adds to and reinforces the speculation of the potential survival in vivo and involvement in pathogenesis of osmotically fragile aberrant forms of bacteria.

Cell Membrane↗

PubMed1978

Complex surveillance of Streptococcus pyogenes. IV. The phenomenon of interference of human serum under the conditions of the bactericidal test.

The phenomenon of interference of human sera under the conditions of the bactericidal test was described. In the presence of these sera, secondarily non-resistant strains of Streptococcus pyogenes were capable of vigorous growth and anti M 12 bactericidal activity was impaired. The factor was found in the albumin fraction and has probably the character of a blocking cytophilic lymphokin. It can markedly distort the interpretation of the bactericidal test.

Adult↗