Streptococcus Type Research Articles

Identification of group B streptococci by immunofluorescence staining.

Gamma globulin fractions of rabbit antisera prepared with whole cell vaccines of group B types Ia, Ib, II, and III and labeled with fluorescein isothiocyanate stained group B streptococci type specifically. Type Ic cells, which contain the Ia polysaccharide antigen of type Ia and the Ic protein antigen of type Ib, were specifically stained by both Ia and Ib conjugates. A group B conjugate pool (B pool) that contained one conjugate specific for each group B type at its predetermined titer gave positive fluorescent-antibody (FA) reactions (4+ intensity) with group B stock strains and negative FA reactions (less than 2+ intensity) with stock strains of streptococcal groups A, C through H, and K through U, viridans streptococci, Streptococcus pneumoniae, Staphylococcus aureus, Neisseria gonorrhoeae, and representative Enterobacteriaceae. Examination of 883 clinical isolates submitted to the Streptococcus Laboratory (Center for Disease Control, Atlanta, Ga.) for identification revealed a 99.1% agreement between FA and culture-precipitin methods. All 305 group B streptococci identified by culture-precipitin and six nonhemolytic group B streptococci missed initially by culture tests were identified correctly by FA. Results of cultural and FA methods in a double-blind study of 99 vaginal swabs agreed on 96 of 99 strains. Three nonhemolytic group B streptococci were identified first by FA and later confirmed by culture-precipitin tests.

Neuraminidase activity in Streptococcus sanguis and in the viridans group, and occurrence of acylneuraminate lyase in viridans organisms isolated from patients with septicemia.

The enzyme neuraminidase (EC 3.2.1.18) was found to be strongly active in different types of Streptococcus sanguis and S. viridans, and, in addition, the occurrence of the enzyme acylneuraminate pyruvate lyase (EC 4.1.3.3) was described in S. viridans. The enzyme-active bacteria strains were isolated from blood cultures of patients with septicemia. Whereas S. sanguis lost its strong neuraminidase activity after some weeks, S. viridans retained its enzyme activity for a long time in culture. Immunoelectrophoretic studies of the blood cultures of patients with streptococcal infections showed the loss of neuraminic acid in most glycoproteins of the serum, proving the in vivo action of neuraminidase. The pathogenic role of neuraminidase is discussed in streptococcal septicemia from the viewpoint of present knowledge.

Identification of Group B Streptococci by Immunofluorescence Staining

Gamma globulin fractions of rabbit antisera prepared with whole cell vaccines of group B types Ia, Ib, II, and III and labeled with fluorescein isothiocyanate stained group B streptococci type specifically. Type Ic cells, which contain the Ia polysaccharide antigen of type Ia and the Ic protein antigen of type Ib, were specifically stained by both Ia and Ib conjugates. A group B conjugate pool (B pool) that contained one conjugate specific for each group B type at its predetermined titer gave positive fluorescent-antibody (FA) reactions (4+ intensity) with group B stock strains and negative FA reactions (less than 2+ intensity) with stock strains of streptococcal groups A, C through H, and K through U, viridans streptococci, Streptococcus pneumoniae, Staphylococcus aureus, Neisseria gonorrhoeae, and representative Enterobacteriaceae. Examination of 883 clinical isolates submitted to the Streptococcus Laboratory (Center for Disease Control, Atlanta, Ga.) for identification revealed a 99.1% agreement between FA and culture-precipitin methods. All 305 group B streptococci identified by culture-precipitin and six nonhemolytic group B streptococci missed initially by culture tests were identified correctly by FA. Results of cultural and FA methods in a double-blind study of 99 vaginal swabs agreed on 96 of 99 strains. Three nonhemolytic group B streptococci were identified first by FA and later confirmed by culture-precipitin tests.

Filamentous capsulated streptococci from the human respiratory tract. I. Antigenic attributes of provisional capsular type 83 and its relationship to streptococci of so-called group M.

Two immunologically reactive polysaccharides have been isolated from the cell walls and from culture filtrates of a filamentous alpha-hemolytic streptococcus provisionally designated capsular type 83. Both polysaccharides were purified by diethylaminoethyl-cellulose chromatography. Analysis indicates that the capsular polysaccharide consists of galactose and phosphorus, whereas the cell wall polysaccharide contains galactosamine, glucosamine, glucose, and phosphorus. On the basis of immunochemical experiments, it is suggested that the capsular polysaccharide is composed of galactose-phosphate units with terminal galactose residues at the nonreducing end. It has also been found that the capsular antigen of streptococcus type 83 is shared by a number of streptococcal strains classified in Lancefield's group M. The cell wall polysaccharide of streptococcus type 83 cross-reacts with antibody to the C(s), or cell wall-like capsular, polysaccharide of Diplococcus pneumoniae, and this cross-reactivity may be a reflection that the streptococcal antigen possesses certain structural features which are similar to those of pneumococcal C and C(s) polysaccharides.

The use of the serum opacity reaction in the typing of group-A streptococci.

SUMMARY A large collection of group-A streptococci was examined for the production of opacity in horse serum and for the presence of M and T antigens. Opacity production is a constant character of certain M types and is consistently absent from the rest. Testing for opacity production is a valuable addition to the serological typing procedure especially if it is carried out first. It has proved useful in (1) reducing the number of tests with M antisera that have to be set up and (2) drawing attention to errors in M typing due to the presence of cross-reacting precipitins in typing sera. The serological specificity of the opacity factor corresponds to that of the M antigen. Inhibition of opacity production therefore provides a useful alternative method of typing strains that form the opacity factor. M antisera for opacity-producing serotypes are often of poor quality and difficult to prepare. In these circumstances, the opacity inhibition test has proved useful for preliminary identification of new M types of streptococci.

EPIDEMIOLOGY OF STREPTOCOCCI IN RYUKYU ISLANDS

Primary and secondary school children in Ryukyu Islands were subjected to throat culture for hemolytic streptococci (HS) to study the incidence of HS as a whole, group and type distribution, and the prevalence of rheumatic heart disease. The results were compared with those obtained in school children in Fuchu City of Tokyo Metropolis by the similar procedure.The incidence of HS as a whole was much higher in Hateruma, Ishigaki and Koza of Ryukyu Islands than in Fuchu City. In typing of group A streptococci by T-agglutination it was interesting to see that the dominant type in Hateruma and Ishigaki was T-11, while in Koza it was T-12, which was similar to the dominat type in Tokyo and other part of Japan. The incidence of rheumatic heart disease in Koza, Ryukyu, was 10 times as high as Fuchu, Tokyo Metropolis, as expected. The discrepancy of types of streptococci in Ryukyu Islands is worth mentioning, and further investigation is continued by us.

An absorption method for preparing anti-M-typing streptococcal sera using acid-hydrolyzed cells.

Abstract The present study describes a procedure for preparing absorption cells that simplifies the preparation of absorbed M-antisera by reducing the number of absorptions necessary to make an antiserum sufficiently specific for use in the typing of streptococci. The method is based on the treatment of the absorbing cells with HCl, thus enchancing the antigenicity of the cells. The method was tested using M-antisera types 11, 12, and 49, and seems to be especially useful for the absorption of those antisera whose absorption by conventional methods is problematic.

Stimulation of Lymphocyte Transformation by Streptococcal Type M1 Protein: Relationship to HL-A Antigens

Abstract M1 protein isolated from group A streptococci, type 1, stimulates human peripheral blood lymphocytes in short-term tissue culture to undergo a dose-dependent increase in DNA synthesis. The degree of this lymphocyte stimulation is not related either to HL-A phenotype, the serum anti-M1 titer of the lymphocyte donor, or the effectiveness of M1 antigen in blocking the cytotoxic activity of HL-A alloantisera. However, both HL-A allo- and heteroantisera are highly effective in specifically inhibiting M1 mitogenic activity. This inhibitory effect clearly depends on anti-HL-A antibody since HL-A alloantisera are rendered ineffective when specifically absorbed with cultured human lymphoid cells. The mechanism by which anti-HL-A antibodies suppress the M1-induced lymphocyte transformation is thought to involve a masking of the M1 protein's mitogenic site such as to prevent interactions with specific lymphocyte receptors required for induction of the proliferative response.

Immune Response in the Mammary Gland

1.Three classes of immunoglobulins have been identified in lacteal secretions. IgG1 appears to be selectively transferred through the glandular epithelium into the udder. Presumably any method of immunization that stimulates circulating antibodies of the IgG1 class will be beneficial, resulting in IgG1 antibodies in the milk.2.IgA class immunoglobulins are also found in the milk, however, in lower concentration than IgG1 immunoglobulins. IgA is produced locally in the gland since an increase is seen in antigen-infused quarters over noninfused quarters. A factor that might be important is that probably the total amount of any immunoglobulin in lacteal secretions will not be increased by systemic immunization whereas local immunization does increase the amount of IgA.3.The method and route of immunization is possibly very important. Intramuscular, subcutaneous, or other routes that result in systemic stimulation of antibodies, to be effective, must for the most part stimulate class IgG1 immunoglobulins. Hammer, Kickhofen, and Henning (7) have recently published that there is a quantitative shift of antibody activity from IgG1 to IgG2 between the first and third courses of immunization of cattle (7). These IgG2 antibodies would be of little value for mammary gland immunity insofar as antibodies in the milk are concerned. However, since the tissues of the mammary gland are involved in a mastitis infection, specific antibodies of all immunoglobulin classes may be beneficial. Immunization by inoculation of a Streptococcus agalactiae, type II preparation into the area of the supramammary lymph node stimulates protective antibodies in the milk. These antibodies have been shown to be of IgG class principally, with less IgA. Finally, by intramammary infusion of antigen into one or more quarters of a nonlactating gland, an increase in specific antibodies of only the IgA class was noted.4.Progress in the determination of the immunogenic substance necessary to stimulate resistance to staphylococcal mastitis has been slow. Serological typing of staphylococci is not available; however, on the basis of phage typing, most mastitis staphylococci may fall into a dozen or so types (14). Serological typing and the immunogenic response of the group B streptococci are much further advanced. The immunology of the group C and group E streptococci is also in a primitive state. It probably is not necessary to separate and purify each immunogen that is required in a preparation; however, it is necessary that inactivated bacteria of each different antigenic type be present in a bacterin in order to stimulate all the various antibodies necessary.One can speculate that a combination of immunization routes that result in the stimulation of several of the classes of immunoglobulins might produce sufficient protective antibodies in the milk and in the mammary gland tissues to increase resistance. Local stimulation would increase specific IgA while systemic stimulation would result in specific IgG1, IgG2 and IgM. Local stimulation would presumably have to occur in the nonlactating gland with a nonirritating or nontissue damaging antigen. The antigen may be a bacterin consisting of a combination of each of the types of staphylococci and streptococci and be inactivated with formalin or some other inactivant. The bacterin should also contain staphylococcal toxoids.

New approaches for the laboratory recognition of M types of group A streptococci.

The successful classification of Group A streptococci by the capillary precipitin technique requires a complete series of M type antisera which are sufficiently potent and specific to give unequivocal type-specific reactions with all the serotypes. Specific antisera for this purpose have been prepared by absorption with heterologous streptococci. Unabsorbed antisera have been employed here in the Ouchterlony double-diffusion agar-gel test to identify the M type of streptococci. Techniques have been developed for making this method of M typing fully reliable. The results reported here confirm and amplify the original findings of Michael and Massell (3). With crude HCl extracts and unabsorbed M type antisera, a precipitin line due to the M protein and another to the group-specific carbohydrate are the two major reactions observed. These reactions, however, are usually readily distinguishable. There was a surprising lack of cross-reactive precipitin lines due to non-type-specific protein antigens in the extracts. Although many of the unabsorbed M type antisera can be employed in the double-diffusion tests, the group-specific antibody must be removed from some of the unabsorbed antisera to avoid confusing cross-reactions. Absorption of these antibodies has been achieved by means of a specific immunoabsorbent column prepared from para-aminophenyl-beta-N-acetylglucosamine and cyanogen bromide-activated Sepharose. Excellent agreement was observed between the M typing results obtained on 117 field strains by the conventional capillary precipitin method and the Ouchterlony double-diffusion method.

Application of Enzyme Production Properties in Subtyping of Group A Streptococci According to T Type

The production of extracellular nicotinamide adenine dinucleotide glycohydrolase (NADG) and the cell-bound lipoproteinase (serum opacity reaction, SOR) by strains of different serological types of group A streptococci, in relation to the T typing, was studied. The production of both NADG and SOR, or only one of them, was found to be characteristic of serotypes, as determined by M and T antigen. No difference in the production of these enzymes was found in relation to M-positive and M-negative variants. Investigation into NADG and SOR production as related to the T type enabled the division of a single agglutination pattern into four main groups, each of which corresponds to one specific M type or more. Of the 370 strains belonging to 12 different T-agglutination patterns, 21% produced both enzymes and 42.5% failed to produce any of them, whereas the remaining 36.5% produced only one out of the two enzymes. Five streptococcal types which did not produce NADG and SOR also failed to synthesize streptolysin S at the early logarithmic phase of growth, indicating that streptolysin S production by young cultures may be also related to serotype. No correlation was found between the production of NADG-SOR as related to serotype and the production of streptolysin O, acid phosphotase, esterase, N -acetylglucosaminidase, hyaluronidase, streptokinase, and the cell-sensitizing factor. The practical and potential usefulness of NADG and SOR production in epidemiological studies is discussed.

Cross-protective antigens of group A streptococci types 3 and 31 and types 46 and 51.

Reciprocal cross-protective antigens have been demonstrated between types 3 and 31 cocci, and one-way cross-protective antigens have been demonstrated between types 46 and 51 cocci. The reciprocal cross-protective antigen of types 3 and 31 is distinct from the specific M protein of either type. In the one-way cross relationship, type 46 cocci contain both type 46 and type 51 M proteins and therefore stimulate protective antibodies against both types. Type 51 cultures contain only the homologous M antigen. These relationships were demonstrated by capillary precipitin tests, indirect bactericidal tests, and in agar-gel diffusion patterns. The practical significance of these relationships in the serological typing of group A streptococci is discussed along with their possible role in immunity to streptococcal infections in man.

Changing types of nephritogenic streptococci in Trinidad.

The relation of seven different M types of streptococci to acute glomerulonephritis associated with skin lesions in South Trinidad has been studied by means of type-specific antibody assays as well as by isolation and identification of the strains. The data indicate that, one after another, five of these strains have prevailed among patients with acute glomerulonephritis during the past five years. At least three of the strains (M-types 55, 49, 57, and/or 60) were associated with epidemic increases in nephritis cases. The appearance of five consecutively predominant types of nephritogenic streptococci during a relatively short period of time is in contrast to the continuing prevalence of M-type 12 strains among nephritogenic streptococci primarily associated with respiratory infections in temperate zones. These observations suggest that the skin sores commonly found on children in tropical Trinidad, provide a particularly suitable environment for development of nephritogenic types. It remains to be seen whether these types will recur or whether new types will continue to emerge in Trinidad.

Application of Enzyme Production Properties in Subtyping of Group A Streptococci According to T Type

The production of extracellular nicotinamide adenine dinucleotide glycohydrolase (NADG) and the cell-bound lipoproteinase (serum opacity reaction, SOR) by strains of different serological types of group A streptococci, in relation to the T typing, was studied. The production of both NADG and SOR, or only one of them, was found to be characteristic of serotypes, as determined by M and T antigen. No difference in the production of these enzymes was found in relation to M-positive and M-negative variants. Investigation into NADG and SOR production as related to the T type enabled the division of a single agglutination pattern into four main groups, each of which corresponds to one specific M type or more. Of the 370 strains belonging to 12 different T-agglutination patterns, 21% produced both enzymes and 42.5% failed to produce any of them, whereas the remaining 36.5% produced only one out of the two enzymes. Five streptococcal types which did not produce NADG and SOR also failed to synthesize streptolysin S at the early logarithmic phase of growth, indicating that streptolysin S production by young cultures may be also related to serotype. No correlation was found between the production of NADG-SOR as related to serotype and the production of streptolysin O, acid phosphotase, esterase, N-acetylglucosaminidase, hyaluronidase, streptokinase, and the cell-sensitizing factor. The practical and potential usefulness of NADG and SOR production in epidemiological studies is discussed.

Further immunological studies of aortitis syndrome.

An elevated titer of anti-aorta antibodies was detected with a significantly higher frequency in cases of aortitis syndrome than in those of other diseases by the complement fixation test and tannic acid hemagglutination test. This finding indicates participation of immune mechanism in the development of aortitis syndrome. The direct fluorescent antibody technique disclosed that aorta antigens are present in the media and the adventitia in a reticular form surrounded by the elastic fibers.A lower titer of anti-aorta antibodies was found with aorta, previously treated with collagenase, than that with untreated aorta, suggesting that collagen might be the main part of the aorta antigen. Boyden's tannic acid hemagglutination technique as modified by Kumagai, was used to study streptococcal type specific antibodies. Most sera from patients with aortitis syndrome showed significantly high titers in one or more types. It is suggested that streptococcal infection may be one of the triggers working in its origin of the development of aortitis syndrome.

A Community-Wide Streptococcal Control Project

A unique, legally enforced, mass-culture program for detection and treatment of streptococcal pharyngitis has been in effect in Natrona County, Wyoming, since 1954. To evaluate the impact of such a program on the streptococcal milieu, the Streptococcal Disease Section, Ecological Investigations Division, Public Health Service, studied a randomized sample of the children under surveillance. The study revealed an extremely low total streptococcal isolation rate (1.8%), as well as antibody levels consistent with a low level of streptococcal infection. The percentage of students with group A antibody was half that of comparable groups in the region; type-specific antibody titers against six streptococcal types revealed values significantly lower than in uncontrolled schoolchild groups in the Rocky Mountain region. The control measures appear to have altered the streptococcal milieu in a favorable manner.

A community-wide streptococcal control project. The Natrona County Primary Prevention Program, Casper, Wyo.

A unique, legally enforced, mass-culture program for detection and treatment of streptococcal pharyngitis has been in effect in Natrona County, Wyoming, since 1954. To evaluate the impact of such a program on the streptococcal milieu, the Streptococcal Disease Section, Ecological Investigations Division, Public Health Service, studied a randomized sample of the children under surveillance. The study revealed an extremely low total streptococcal isolation rate (1.8%), as well as antibody levels consistent with a low level of streptococcal infection. The percentage of students with group A antibody was half that of comparable groups in the region; type-specific antibody titers against six streptococcal types revealed values significantly lower than in uncontrolled schoolchild groups in the Rocky Mountain region. The control measures appear to have altered the streptococcal milieu in a favorable manner.

Endemic Nephritis and Streptococcal Infections in South Trinidad

Surveillance of streptococcal infections has been continued in South Trinidad following a major epidemic of acute glomerulonephritis. Observations made during the first postepidemic year (July 1966 to July 1967) are described in the present report. Skin lesions, streptococcal infections, and serum antibody titers are compared in 148 patients with nephritis and 101 clinically well school children as well as in 882 members of patients' families and 667 members of families without nephritis. Skin lesions were as common among the "well" school children as they had been during the epidemic while streptococci were isolated even more frequently from these lesions. However, prevalent types of streptococci differed from those found in 1965. Antistreptolysin O titers were less often increased in patients while antihyaluronidase titers continued to be elevated in all groups. Anti-group A carbohydrate titers were increased in patients.

Beta-hemolytic streptococcal and concurrent infections in adults and children with respiratory disease, 1958 to 1969.

Surveillance of respiratory disease in adults and children from various populations in Louisiana and Mississippi from 1958 to 1969 revealed that 7.1 per cent of adults and 9.3 per cent of children had beta-hemolytic streptococci in their pharyngeal cultures at onset of illness. In adults, one third of the streptococci were Group A and two-thirds were Groups B, C, and G whereas in children this distribution was reversed. The type of streptococci present could not be differentiated by clinical criteria. There were no epidemic strains and no sizeable outbreaks of infection. The greatest percentage (18 per cent to 25 per cent) of cultures yielding streptococci occurred in patients with pharyngitis, but all clinical types of respiratory disease were included. Evidence of concurrent infection by viruses or Mycoplasma pneumoniae and beta-hemolytic streptococci was found frequently, but there was no pattern of association with a particular agent and no evidence of enhancement of illness.

Identification of types of potentially cariogenic streptococci on human tooth surfaces by the fluorescent antibody technique.

The frequency distribution of four potentially cariogenic streptococcal types, AHT, BHT, GS-5, and HHT, on seven specific maxillary and mandibular tooth surface segments of 27 male teenagers was investigated. BHT and HHT serotypes were widespread in the dentition of all subjects. AHT and GS-5 organisms were identified in 7 and 89% of the population studied, respectively.