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Biomedical subjects

J M Fournier

Publications and source records attributed to J M Fournier.

At least 55 records · Page 3Linked to original sources
PubMed1991

Production of type 5 capsular polysaccharide by Staphylococcus aureus grown in a semi-synthetic medium.

The concentration of the type 5 capsular polysaccharide (CP) antigen of Staphylococcus aureus can be measured directly in cultures or cell suspensions by a two-step inhibition enzyme-linked immunosorbent assay (ELISA), using monoclonal antibodies. CP was synthesized during growth on a variety of carbon substrates and its production was not affected by the nature of the carbon source. High levels of yeast extract inhibited CP formation. CP was synthesized in batch culture at the same rate during exponential growth as in the post-exponential phase. Post-exponential CP production contributed at least half the final amount of CP measured. This phenomenon was observed in different culture media, although the specific yield of polysaccharide varied from one medium to another. Post-exponential CP production was observed in the pH range 6-7, but not at pH 8. Post-exponential production was strictly dependent on oxygen availability and did not occur under anaerobic conditions.

Antigens, Bacterial↗
PubMed1991

Staphylococcus aureus growth and type 5 capsular polysaccharide production in synthetic media.

The production of type 5 capsular polysaccharide by Staphylococcus aureus in synthetic media was investigated. The influence of medium components on capsular polysaccharide synthesis appeared to relate to the presence or absence of the component rather than to concentration gradient. The production of type 5 capsular polysaccharide was linked to energy availability and energy source, but not to carbohydrate concentration or carbon/nitrogen ratio. Regulation of capsular polysaccharide production by S. aureus in response to medium changes would appear to differ from that typically displayed in other organisms that produce polysaccharides.

Bacterial Capsules↗
PubMed1991

Production of polyclonal and monoclonal antibodies against group A, B, and C capsular polysaccharides of Neisseria meningitidis and preparation of latex reagents.

Polyclonal and monoclonal antibodies against capsular polysaccharides of Neisseria meningitidis serogroups A, B, and C were produced in order to develop immunological reagents allowing both the detection of soluble antigens during meningococcal meningitis and antigenic serogrouping of N. meningitidis cultures. The performance characteristics of monoclonal and polyclonal antibody latex reagents were compared. For the detection of soluble polysaccharide antigen, polyclonal antibody latex reagent was selected for N. meningitidis A and C. The latex reagent prepared with polyclonal antibodies against N. meningitidis B could not detect capsular polysaccharide even at 1 mg/ml. The monoclonal antibody B latex reagent which detected 100 ng of polysaccharide per ml was therefore chosen. For the serogroup identification of N. meningitidis, the use of a confirmatory test results in an overall specificity of 100% with polyclonal or monoclonal antibody latex reagents.

Antibodies, Bacterial↗
PubMed1991

Comparison between lactoferrin and subepithelial matrix protein binding in Staphylococcus aureus associated with bovine mastitis.

Staphylococcus aureus strains (n = 100) isolated from bovine mastitis were classified according to the presence of capsular polysaccharide serotype 5 (n = 46), type 8 (n = 26), and non-5/8 (n = 28). Strains from each type were tested for protein interaction in a 125I-labeled ligand binding assay. A majority of type 5 and type 8 strains showed a higher degree of binding to lactoferrin, fibronectin, and IgG than the non-5/8 strains. Fibrinogen binding was low in all serotypes. Most of the type 5 and non-5/8 strains bound less than 10% laminin, whereas type 8 strains bound laminin in the 11 to 20% range. Non-5/8 strains significantly differed from type 5 in lactoferrin, fibronectin, fibrinogen, and IgG and also from type 8 in fibrinogen and IgG binding. The differences in protein binding between type 5 and type 8 were nonsignificant. The degree of lactoferrin binding in all types positively correlated with laminin binding. Lactoferrin and fibrinogen bindings were correlated in type 5 and type 8 strains. Lactoferrin and fibronectin bindings were correlated only in type 5 strains. These data suggest that bovine lactoferrin binding is common and associated with subepithelial matrix protein interactions in certain serotypes of S. aureus.

Animals↗
PubMed1990

Structure of the type 5 capsular polysaccharide of Staphylococcus aureus.

The Staphylococcus aureus type 5 capsular polysaccharide is composed of 2-acetamido-2-deoxy-L-fucose (1 part), 2-acetamido-2-deoxy-D-fucose (1 part), and 2-acetamido-2-deoxy-D-mannuronic acid (1 part). On the basis of methylation analysis, optical rotation, high-field one- and two-dimensional 1H- and 13C-n.m.r. experiments, and selective cleavage with 70% aqueous hydrogen fluoride, the polysaccharide was found to be a partially O-acetylated (50%) polymer of the repeating trisaccharide unit, [----4)-3-O-Ac-beta-D-ManpNAcA-(1----4)-a-L-FucpNAc-(1----3) -beta-D-FucpNAc-(1----]n.

Carbohydrate Sequence↗
PubMed1990

Correlation between esterase electrophoretic types and capsular polysaccharide types 5 and 8 among methicillin-susceptible and methicillin-resistant strains of Staphylococcus aureus.

The relationship between capsular polysaccharide types 5 and 8 and esterase electrophoretic types (zymotypes) in 160 French clinical isolates of Staphylococcus aureus was studied. Methicillin-susceptible strains of capsular types 5 and 8 were represented by 11 zymotypes, indicating a high polymorphism. Methicillin-resistant strains were mainly distributed in only two distinct populations. The predominant population was represented by strains of zymotype 6 and capsular type 5, and the second population was represented by strains of zymotype 14 and capsular type 8.

Bacterial Capsules↗
PubMed1990

Reactivity of coagulase-negative staphylococci isolated from cow and goat milk with monoclonal antibodies to Staphylococcus aureus capsular polysaccharide types 5 and 8.

Monoclonal antibodies to Staphylococcus aureus capsular polysaccharide types 5 and 8 were used in an enzyme-linked immunosorbent assay to serotype 74 and 42 coagulase-negative isolates from cow and goat milk, respectively. Eighteen (15.5%) isolates were typable: 13 Staphylococcus haemolyticus, 1 S. hyicus, 1 S. simulans, and 1 S. warneri from bovine origin and 2 S. lentus from caprine origin. Type 5 was predominant, accounting for about 89% of typable isolates. Reactivity with monoclonal antibodies varied considerably according to isolates. The significance and the potential importance of these findings are discussed.

Animals↗
PubMed1990

Epidemiological markers for epidemic strain and carrier isolates in an outbreak of nosocomial oxacillin-resistant Staphylococcus aureus.

An outbreak of nosocomial infections occurring in a postoperative intensive care unit was caused by a single strain of oxacillin-resistant Staphylococcus aureus. Six patients were infected, or colonized, by this strain, which was traced by using the following four epidemiological markers: antibiogram, bacteriophage type, capsular polysaccharide type, and esterase electrophoretic type. This strain was compared with S. aureus isolates obtained from the noses of 13 carriers from a group of 42 staff members. A good correlation in terms of phenotypic markers was found between the epidemic strain and a strain isolated from one carrier. Both exhibited the same pattern of multiple resistance as well as the same phage type, 77, capsular polysaccharide type, 5, and esterase electrophoretic type, 6. In contrast, an oxacillin-resistant strain, isolated from another carrier, differed from the epidemic strain by susceptibility to rifampin and by susceptibility to four additional bacteriophages. The other 11 strains isolated from carriers were susceptible to oxacillin and exhibited widely different phenotypes. These results confirm the interest of using several epidemiological markers to trace the spread of epidemic S. aureus strains and to delineate the carrier strains.

Bacterial Typing Techniques↗
PubMed1989

Mucoid phenotype of Klebsiella pneumoniae is a plasmid-encoded virulence factor.

We have previously reported that the presence of a 180-kilobase plasmid encoding production of aerobactin was correlated with the virulence of Klebsiella pneumoniae K1 and K2 isolates. This work demonstrates that a variant of a K2 strain which has lost this plasmid, pKP100, becomes avirulent. Labeling of this plasmid with the mobilizable, replication-defective element pME28, used here as a mobilizable transposon, allowed the transfer of this plasmid into a plasmidless derivative. Virulence was restored upon reacquisition of this tagged plasmid, pKP101. In addition to aerobactin production, another phenotype could be correlated with the presence of this virulence plasmid: the mucoid phenotype of the bacterial colonies. Both wild-type and plasmidless strains are encapsulated, but only the former presented mucoid colonies. Participation of this phenotype in the virulence of K. pneumoniae was demonstrated by constructing a mutant altered in the plasmid gene encoding this phenotype. The resulting strain demonstrated a 1,000-fold decrease in virulence. Introduction of the recombinant plasmid pKP200 carrying the gene encoding this mucoid phenotype into Escherichia coli HB101 also led to the production of a mucoid phenotype. Rocket immunoelectrophoresis demonstrated that in E. coli this phenotype was due to the production of colanic acid. On the other hand, neither the overproduction of K2 capsular polysaccharide nor the presence of colanic acid was detected in mucoid strains of K. pneumoniae. We conclude that this mucoid phenotype is definitely an important virulence factor of K. pneumoniae. It is due to the plasmid-encoded production of a substance which is different from colanic acid and the capsular polysaccharide of K. pneumoniae.

Base Composition↗
PubMed1989

Direct testing of blood culture for detection of the serotype 5 and 8 capsular polysaccharides of Staphylococcus aureus.

Monoclonal antibodies (MAbs) reactive with serotype 5 and 8 capsular polysaccharides of Staphylococcus aureus have been used to test, by enzyme-linked immunosorbent assay (ELISA), blood culture fluids for the presence of S. aureus. A total of 748 blood cultures from 665 patients yielding 706 bacterial isolates belonging to more than 26 bacterial species were studied. All blood cultures containing bacterial strains belonging to species other than S. aureus were negative in ELISA. All 23 blood cultures containing serotype 5 S. aureus were positive in ELISA with the corresponding MAb. Out of 20 blood cultures containing serotype 8 S. aureus, 19 were positive with the corresponding MAb. All 5 blood cultures containing nontypeable S. aureus were negative in ELISA with both MAbs. This method provides reliable identification of serotype 5 or serotype 8 S. aureus by direct testing of blood culture fluids with ELISA.

Animals↗
PubMed1989

Staphylococcus aureus strains which are not identified by rapid agglutination methods are of capsular serotype 5.

A total of 183 recent Staphylococcus aureus clinical isolates were tested with three commercially available rapid agglutination methods. The capsular polysaccharide type and resistance to oxacillin of these isolates were also determined. Seven isolates were not identified correctly by agglutination methods. All isolates not identified by the rapid methods were of capsular serotype 5, and of these isolates, six were resistant to oxacillin. The results suggest that these agglutination kits can be improved by the use of antibodies reactive with S. aureus capsular polysaccharide.

Agglutination Tests↗
PubMed1988

Prevalence of capsular polysaccharide types 5 and 8 among Staphylococcus aureus isolates from cow, goat, and ewe milk.

Monoclonal antibodies to Staphylococcus aureus capsular polysaccharide types 5 and 8 were used to serotype by enzyme-linked immunosorbent assay 212, 54, and 33 isolates from cow, goat, and ewe milk, respectively. Capsular types 5 and 8 accounted for 69.4% of bovine isolates and 71.5 and 78.8% of goat and ewe isolates, respectively. Type 5 was predominant in strains from bovine sources (51.4%), whereas type 8 was prevalent in strains from caprine (68.5%) and ovine (75.8%) sources.

Animals↗
PubMed1988

Staphylococcus aureus capsular types and antibody response to lung infection in patients with cystic fibrosis.

Chronic respiratory tract infections caused by Staphylococcus aureus are common in patients with cystic fibrosis (CF). Recently, it was shown in a few CF patients that S. aureus isolates produce capsular polysaccharides (CPs). However, it is not known whether this is a common feature and whether an immune response to CPs in CF is detectable. Therefore, we examined 170 S. aureus isolates from CF patients and healthy individuals for production of CP types 5 and 8 by using monoclonal antibodies. We found that CP-producing staphylococcal isolates were randomly distributed among CF patients and healthy carriers. Eighty-five percent of all isolates produced CPs, 77% of which were type 8. Examination of one sputum sample by an immunofluorescence technique suggested that production of CPs is not an in vitro phenomenon. S. aureus isolates from various sites of a single person often yielded more than one CP type. A random distribution of S. aureus strains with CP type 5 or 8 from the skin and respiratory tracts of patients and from the skin of healthy individuals was found. Antibody response to CP types 5 and 8, measured by enzyme-linked immunosorbent assay, was not elevated in CF patients with chronic S. aureus lung infection in comparison with healthy carriers. On the contrary, in CF patients the ratios of antibodies to CP 8 were significantly lower (P less than 0.005; alpha = 0.025). The ratios of antibodies to CP types did not change when monitored longitudinally over several months. This study suggests that the production of CPs is a universal property of S. aureus and that infected CF patients do not have elevated ratios of antibodies to these antigens.

Antibodies, Bacterial↗
PubMed1988

[Influence of different types of experimental obesity on resistance of the mouse to infection by Salmonella typhimurium and Klebsiella pneumoniae].

Resistance to infections inducing two types of immune response, humoral and cell-mediated, has been measured in mice after Salmonella typhimurium and Klebsiella pneumoniae inoculation; the animals exhibited different kinds of obesity: genetic, ob/ob and db/db mutants, induced by fat diet or gold thioglucose (aurothioglucose) injection (determining obesity of central origin). Klebsiella infection was aggravated in all types of obesity. Salmonella infection was aggravated in genetically diabetic and dietary-obese mice. The two kinds of genetically obese mice show an important functional decrease in splenic lymphocytes. In contrast, aurothioglucose-obese mice were more resistant than controls.

Animals↗
PubMed1988

[Detection of heterogeneous Staphylococci by the Autobac system using a nafcillin disk].

Detection of Methicillin Resistant Staphylococcus aureus (MRSA) is still a problem for automated system users. Forty-nine strains were studied--two kinds of tests were performed: the first by using a 1.0 microgram nafcillin disk with the Autobac system, the second by testing an oxacillin disk by diffusion on Mueller-Hinton agar base complemented with natrium chloride incubated at 37 degrees C. Between the two tests, we observed a concordance for 87.8% of the strains. Six strains showed a discrepancy with the reference method. But for five of them, the Light Scattering Index (LSI) showed a decreasing sensibility near for the cut off. Whereas, only one strain is considered as susceptible. The 1.0 micrograms nafcillin disk allow better results than the 5.0 micrograms oxacillin disk in the automated system, Autobac.

Automation↗
PubMed1987

Predominance of capsular polysaccharide type 5 among oxacillin-resistant Staphylococcus aureus.

The relationship between capsular polysaccharide types 5 and 8 and resistance of Staphylococcus aureus to oxacillin was studied with a collection of 406 clinical isolates from six French hospitals. Of 175 type 5 isolates, 84 (48%) were resistant to oxacillin. In contrast, only 8 of 160 type 8 isolates (5%) and 5 of 71 nontypeable isolates (7%) were resistant to oxacillin. Therefore, capsular typing of clinical isolates of S. aureus may facilitate the choice of first-line antibiotic therapy.

Bacterial Capsules↗
PubMed1986

Immunogenic properties of Klebsiella pneumoniae type 2 capsular polysaccharide.

The immunoprotective activity of Klebsiella pneumoniae K2 cell surface preparations and purified capsular polysaccharide was tested in mice. The 50% protective dose (PD50), expressed as capsular polysaccharide content, was 2 ng for cell surface preparations and 50 ng for purified capsular polysaccharide. Both preparations lost their immunoprotective activity after alkali treatment. Immune sera were raised in rabbits immunized with cell surface preparations. The precipitating and hemagglutinating capacity of these antisera was tested against either purified capsular polysaccharide or alkali-treated capsular polysaccharide. No difference was observed between the reactivity of the antisera against each antigen. The protective activity of these sera was tested on mice in passive transfer experiments, before and after absorption with either purified capsular polysaccharide or alkali-treated capsular polysaccharide. The sera lost their protective activity after absorption with purified capsular polysaccharide and after absorption with alkali-treated capsular polysaccharide. These experiments show that the difference in immunoprotective activity of cell surface preparations, purified capsular polysaccharide, and alkali-treated capsular polysaccharide is not due to a difference in their antigenic determinants. Cell surface preparations and purified capsular polysaccharide were fractionated by gel filtration on Sepharose 4B and by ultracentrifugation on cesium chloride density gradients. Three forms of capsular polysaccharide have been characterized. (i) A form of capsular polysaccharide with a very high protective activity (PD50 = 2 ng) that copurified with protein and lipopolysaccharide and was characterized by a low coefficient of distribution (Kd = 0.20) and a low density (1.5 to 1.6 g/cm3). (ii) A form of capsular polysaccharide with an intermediate protective activity (PD50 = 50 ng), contamined by less than 3% protein and 1% lipopolysaccharide, with a Kd of 0.35, and a density of 1.7 to 1.8 g/cm3. (iii) A nonimmunoprotective capsular polysaccharide obtained after alkali treatment of either cell surface preparations or purified capsular polysaccharide. The Kd of these fractions varied from 0.20 to 0.90 and their density from 1.7 to 1.8 g/cm3.

Animals↗