Streptolysin S: purification and properties.
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Carrier Proteins↗
Biomedical subjects
L W Wannamaker
Publications and source records attributed to L W Wannamaker.
Studies of the receptor for phage A25 in group A streptococci: the role of peptidoglycan in reversible adsorption.
Irreversible adsorption of a virulent phage, phage A25, to heat-killed streptococci, groups A, G, and A variant, has been achieved. Adsorption reflected the observed host range for phage A25 in that heat-killed group B cells were not able to inactivate the phage. Broken cells, cell walls, and peptidoglycan prepared from a group A strain K56 failed to adsorb the phage irreversibly, but retained the potential to carry out reversible adsorption. Experimental data including electron microscopy have demonstrated the specificity of reversible adsorption and have identified the peptidoglycan as a necessary cellular component of the receptor. The sensitivity of whole cells and purified peptidoglycan to muralytic enzymes suggests that the cell wall and peptidoglycan must be intact for optimal adsorption. In general the results are explained by postulating that adsorption of A25 phage particles to group A cells occurs by a two-step process; the first step involves recognition and reversible binding of the phage tail to the cell wall peptidoglycan, the second step is an irreversible reaction catalyzed by a yet unidentified cellular component which is destroyed when cells are ruptured.
Adsorption↗
Hyaluronidase activity of bacteriophages of group A streptococci.
A sensitive dye-binding assay was employed to study the hyaluronidase associated with temperate and virulent phages infected group A streptococci. Some enzyme was detectable in each purified phage preparation examined, but differences of several orders of magnitude separated the lower enzyme levels in virulent phages that required the addition of hyaluronidase for plaque formation and the higher levels in temperate phages that did not. Infection by virulent phage A25 was accompanied by the production of levels of hyaluronidase proportionate to the average burst size. Hyaluronidase was produced during infection by temperate phages at a much higher level than could be accounted for by the number of phage particles formed. The major portion of this hyaluronidase was free and apparently unassociated with phage or phage fragments. The phage-associated enzyme was tightly bound but could be released and solubilized by treatment with urea.
Bacteriophages↗
Typing of Group A streptococci from an urban area (Köln) in West Germany.
Two-hundred and twenty strains of Streptococcus pyogenes (group A streptococcus) from the urban area of Köln, West Germany have been typed by M precipitin and T agglutination tests. Determination of streptococcal nicotinamide adenine dinucleotide glycohydrolase (NADase) production and the serum opacity reaction (SOR) have also been performed. 65.5% of the isolates could be typed by the M precipitin reaction. 92.1% of the remaining strains could be identified by their T protein antigens and could be further subdivided according to their NADase/SOR pattern. Most of the M typable strains originated from upper respiratory tract infections (including scarlet fever), whereas the majority of the M nontypable streptocci had been isolated from skin, wound, and other non-respiratory sites.
Agglutination Tests↗
C-reactive protein in streptococcal pharyngitis.
This study was designed to explore whether the test for C-reactive protein (CRP) is useful in differentiating bona fide streptococcal infection from the symptomatic carrier at the time of the acute visit to the physician. Serial blood samples from 157 children with symptomatic pharyngitis and a positive culture for group A streptococci were analyzed for the presence or absence of CRP. These data were compared with the patients' antibody responses to two streptococcal extracellular antigens (antistreptolysin O and antistreptococcal deoxyribonuclease B). Seventy-eight percent of patients with serologically confirmed streptococcal pharyngitis had a positive CRP test at the initial visit. Conversely, if the CRP test was negative at the acute visit, only about 25% later showed an antibody response. This latter finding held regardless of the degree of positivity of the initial culture, the presence of exudate or adenitis, or the presence of a temperature greater than 38.3 C (101 F) or coryza. These data suggest that the CRP test may be helpful to the clinician, especially if this abnormal protein is absent at the time of the acute visit.
C-Reactive Protein↗
A.H.A. committee report. Prevention of rheumatic fever.
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Anti-Bacterial Agents↗
Suppression of the antistreptolysin O response by cholesterol and by lipid extracts of rabbit skin.
Lipids extracted from rabbit skin block the hemolytic capacity of SO and also suppress the neutralizing antibody response to this streptococcal extracellular antigen in rabbith immunized intravenosly. The modification in antibody response is specific for SO; the antibody responses to streptococcal DNase B and to streptococcal NADase are not affected. Cholesterol, a lipid present in abundance in skin, has a similar specific effect on the antigenicity of SO and may be the component responsible for the demonstrated effects of these lipid extracts of skin. In vitro experiments indicate that lipid extracts of rabbit skin have a greater capacity to block the hemolytic capacity of SO than do lipid extracts of rabbit heart, kidney, lung, liver, or spleen. These data support the view that the feeble ASO response observed in patients with streptococcal pyoderma is a result of the abundance of a local lipid inhibitor, such as cholesterol, in the skin. They may also bear on the pathogenesis of rheumatic fever, a complication which apparently does not occur following group A streptococcal pyoderma. Two possible explanations for this remarkable epidemiologic observation, both related to the presence of a local inhibitor, are considered: (a) suppression of the ASO response, the magnitude of which has been correlated with the risk of developing rheumatic fever after streptococcal infection of the throat, and (b) inhibition of the toxicity of SO, which has been shown to have a direct toxic effect on the mammalian heart and on isolated beating myocytes.
Animals↗
[Antibody response to group A streptococcal exoenzymes (author's transl)].
Sera from normal controls from patients with streptococcal diseases, and from other patients whose serum was sent for anti-streptolysin 0 determinations were tested for antistreptolysin 0 (ASO) and anti-DNase B (ADB) antibodies. The Streptozyme test was performed on the same sera. The upper limits of normal in the control population were established as 160 units for ASO and as 240 units for ADB, respectively. The usefulness of the ADB test in addition to the ASO test was confirmed: when both tests were performed elevated titers could be demonstrated in a higher percentage of various streptococcal diseases. With respect to streptococcal infections of the skin the anti-DNase B-test was superior to the ASO test. In this study of the Streptozyme test showed an elevated antibody titer in a lower proportion than the other two tests.
Adolescent↗
Transduction of bacteriocin determinants in group A streptococci.
Determinants of streptococcin A-FF22 (SA) production and host cell immunity have been transduced to three serologically distinct Group A streptococci. Streptomycin resistance markers were not cotransducible with bacteriocin determinants. SA+ transductants of strains unrelated to the parent SA+ strain were unstable but SA+ transductants of a spontaneous SA- derivative of the parent appeared to be stable.
Bacteriocins↗
A penicillin shot without culturing the child's throat.
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Carrier State↗
The dynamics of streptococcal infections in a defined population of children: serotypes associated with skin and respiratory infections.
Serial observations including cultures of the upper respiratory tract and of infected skin lesions and streptococcal antibody determinations were made over a two-year period in a semi-closed population of children between 10 months and 15 years of age. There was a high prevalence of group A streptococci in nose and throat cultures and of skin lesions containing these organisms. Almost 90% of the study population developed streptococcal impetigo during the study period. A slightly higher proportion of males than females developed skin infection but there was no relationship to age. Impetigo was observed throughout the calendar year, exceeding 12% of child-visits in one winter month, but was generally more common in the summer and fall. Conversely, group A streptococci were more often isolated from nose and throat cultures in the winter months. The increase in recovery of streptococci from nose and throat cultures lagged behind the increase in streptococcal impetigo and continued into the winter months, when the prevalence of impetigo had declined. Calculation of ratios for individual streptococcal serotypes isolated from different body sites revealed a clear cut distinction between "respiratory" and "impetigo" serotypes, with respect to both prevalence and acquisition rates. Respiratory serotypes were more commonly isolated in the winter and impetigo serotypes in the summer and fall. Significant antibody responses to extracellular antigens of the streptococcus were documented for pharyngeal acquisitions of both impetigo and respiratory serotypes and for skin lesions associated with impetigo serotypes. Group A streptococcal serotypes may be divided into three categories on the basis of their human pathogenicity for body sites: some with the potential for respiratory infection, others with the potential for skin infection and a few unusual serotypes with the potential for infecting both sites.
Age Factors↗
Genetic basis of streptococcin A-FF22 production.
Spontaneous, low-frequency loss of ability to produce streptococcin A-FF22 (SA) by group A streptococcus strain FF22 was observed. The proportion of non-SA-producing (SA(-)) derivatives occurring in strain FF22 cultures grown in Todd Hewitt broth supplemented with 1% of yeast extract (THBY) was increased on treatment with ethidium bromide, acriflavin, or rifampin. The highest incidence of SA(-) organisms, however, was found in untreated THBY cultures that had been aging by incubation at 37 degrees C for several months. The possibility of selective effects in these experiments, operating to enhance the apparent frequency of SA(-) bacteria, was discounted. The survival of SA(-) derivatives in association with populations of SA(+) bacteria was dependent upon the use of culture conditions inimical to SA activity, since a consistent finding was that the loss of ability to produce SA was associated with loss of immunity to the killing action of this bacteriocin. Whereas selective killing of SA(-) derivatives was evident in mixed cultures of SA(+) and SA(-) strains in tryptic soy broth, no such effect was demonstrable in THBY. In these experiments, elimination of SA(-) cells seemed directly related to the presence of active SA. Purified clones of SA(-) substrains did not seem revertible to SA production, either spontaneously or on treatment with nitrosoguanidine. It is suggested that the property of production of SA by group A streptococcus strain FF22, together with that of host cell immunity to the homologous bacteriocin, may be mediated by plasmid-borne genetic determinants.
Bacteriocins↗
Bacteriocins of gram-positive bacteria.
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Antibiosis↗
Application of a new method for detecting streptococcal nicotinamide adenine dinucleotide glycohydrolase to various M types of Streptoccus pyogenes.
The production of extracellular nicotinamide adenine dinucleotide glycohydrolase (NADase) by Streptococcus pyogenes can easily be demonstrated using the fluorescence of nicotinamide adenine dinucleotide in ultraviolet light occurring on addition of strong alkali. The new method described here uses microtiter plates and can be read with the naked eye. It permits the screening of large numbers of strains in a short time and with minimal amounts of reagents. The sensitivity of the new method proved to be good in comparison with the bisulfite spectrophotometric method. Culture supernatants of 177 group A streptococci were tested for NADase production by the microtiter fluorescence method. We could confirm former findings that strains within a certain M type of S. pyogenes are usually either producers or nonproducers of the enzyme. The usefulness of the test for screening of streptococcal NADase production is discussed.
Cell-Free System↗
Comparison of three methods for grouping streptococci.
Two new methods for serological grouping of beta-hemolytic streptococci, the nitrous acid extraction procedure of El Kholy et al. and the slide agglutination method of Christensen et al., were compared with the Lancefield hot-hydrochloric acid extraction method in classifying 92 strains of groups A, B, C, and G. The nitrous acid extraction method was easily performed, specific, and sensitive when highly potent antisera were used. For the Christensen method these highly potent antisera had to be diluted to avoid cross-reactions between groups A and C and groups B and G, respectively. A few strains, most of them group B, could not be grouped by the latter method. Using these three grouping methods, two sets of commercial sera were compared with the more potent sera supplied by R. C. Lancefield. The low antibody content of these commercial sera, especially anti-group B and G sera, contributed to the inferior results obtained in some of the grouping reactions.
Agglutination Tests↗
Method for phage typing group A type 49 streptococci.
A method of phage subtyping group A type 49 streptococci is described. The method is similar to that used for phage typing staphylococci, except that lysates obtained by induction with mitomycin C rather than propagated stock phages were used. Five type 49 strains were used as phage donors. Seventy-two strains of type 49 streptococci isolated from 10 worldwide sources were examined by this method. Among these strains, five distinct subtypes (I through V) could be distinguished on the basis of their lytic patterns. Only a few of the type 49 strains could not be classified into one of these phage subtypes (6% using 100 X routine test dilution). Strains from a single source were generally homogeneous with respect to their phage subtype. The method proved useful in discriminating between type 49 strains isolated from different geographical sources and from the same place in different years. Studies in progress suggest that it may be useful for subtyping other strains of special epidemiological interest, such as strains of other serological types associated with nephritis.
Bacteriophage Typing↗
Enumeration of beta-haemolytic streptococci on normal skin by direct agar contact.
Normal skin sites in children from a population in which streptococcal impetigo is endemic were examined for the presence of beta-haemolytic streptococci by a direct agar-contact technique. Ninety-eight of 554 samples (18%) were positive for these organisms. Penicillin prophylaxis reduced the frequency of isolation of streptococci from the normal skin for a period of 3 weeks, perhaps accounting in part for the lower isolation rate in this than in earlier studies. Numbers of streptococcal colony-forming units in positive samples were generally low, both in terms of absolute numbers isolated from the surface area sampled and in comparison with numbers of other aerobic flora recovered. The presence of streptococcal pyoderma at the time of agar contact was not necessarily associated with the presence of or with increased numbers of streptococci on samples obtained from normal skin sites. Low counts were consistently found in early summer and higher counts in some samples in late summer. In a simultaneous comparison of paired samples taken from adjacent sites, the frequency of detection of streptococci by direct agar contact compared favourably with that obtained with a moist-swab method. The increased frequency of detection by the agar-contact method appeared to be related to an increased sensitivity for the detection of low numbers of streptococcal colony-forming units on the normal skin.
Adolescent↗
Bacteriocin of a group B streptococcus: partial purification and characterization.
The production of bacteriocin-like inhibition by two of 135 strains of group B streptococci was demonstrated during the growth of these organisms on solid nutrient media. Although a variety of gram-positive organisms was susceptible, none of a wide range of different gram-negative strains was inhibited. The active substance produced by one of these strains was extracted from cultures grown on solid media (Todd-Hewitt agar) and was shown to be bactericidal in action. This bacteriocin (streptocin B(1)) was partially purified by ammonium sulfate precipitation followed by gel filtration and ion-exchange chromatography. Production of the bacteriocin was repressed by addition of certain fermentable carbohydrates to the basic medium and enhanced by the addition of yeast extract. The bacteriocin was shown to exist in two distinct and interconvertible physical forms: a basic unit of molecular weight 10,000 and an aggregate having a molecular weight of over 200,000. The bacteriocin was inactivated by proteolytic enzymes and, although labile in alkali, it was stable to boiling in mild acids.
Bacteriocins↗