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Teichoic acids in pathogenic Staphylococcus aureus.

Twenty-six strains of Staphylococcus aureus obtained from patients with endocarditis were studied for the production of alpha- and/or beta-ribitol teichoic acid (TA), using highly specific anti-TA antibodies prepared in rabbits. A counterimmunoelectrophoretic assay was used. Beta-TA was the predominant residue produced by all strains; alpha-TA was found in all strains, but in smaller amounts and with much strain-to-strain variations. Antibodies in patients' sera were found against beta-TA in higher titers and for longer periods than were anti-alpha-TA antibodies. Antibodies against one or both TA residues were present in all but one of 26 patients.

Antibodies, Bacterial↗

[Comparative immunochemical characteristics of extracellular polysaccharide-containing preparations and teichoic acids of Staphylococcus aureus].

The sensitivity of the enzyme immunoassay on the basis of polysaccharide-containing preparations was found to be lower than that of the assay on the basis of teichoic acids. The specificity of both assays was 100%. There was no complete coincidence between the results of the detection of antibodies to these two antigens in patients' sera.

Animals↗

Technical aspects of the Staphylococcus aureus teichoic acid antibody assay: gel diffusion and counterimmunoelecrophoretic assays, antigen preparation, antigen selection, concentration effects, and cross-reactions with other organisms.

Because variable results are being reported from laboratories performing the teichoic acid antibody assay in patients with serious infections due to Staphylococcus aureus, we have thoroughly reviewed all technical aspects of the test. This paper reports on the importance of agar and antigen preparation in standardizing results of the assay and reducing the prevalence of false-negative and -positive tests. Once standardized, the counterimmunoelectrophoretic method is as accurate as the gel diffusion method for both initial screening and generating titers; practically, however, unless numerous tests are to be performed, the gel diffusion technique will suffice for most purposes. The cell wall of the Lafferty strain of S. aureus was used as the standard antigen in assays for serum antibodies. We studied whether it is an appropriate antigen and found that antibody titers obtained with the Lafferty strain antigen were, in three patients with endocarditis, the same as those obtained with antigens from the individual blood stream isolates. We have also confirmed that pooled human gamma globulin can be used, by back titration against newly prepared lots of antigen, to select optimal antigen concentration and is as good as more specific, higher titer serum specimens for that purpose. Finally, cell wall antigens from Staphylococcus epidermidis and a variety of streptococci may react with normal human sera, but such antigens are distinct by immunoprecipitation from those from S. aureus.

Antibodies, Bacterial↗

Studies on the linkage between teichoic acid and peptidoglycan in a bacteriophage-resistant mutant of Staphylococcus aureus H.

1. In addition to poly(ribitol phosphate) the walls of a bacteriophage-resistant mutant of Staphylococcus aureus H contain glycerol phosphate residues that are not removed on digestion with trypsin or extraction with phenol. 2. The glycerol phosphate is present in a chain, containing three or four glycerol phosphate residues, which is covalently attached to the peptidoglycan through a phosphodiester linkage to muramic acid; this linkage is readily hydrolysed by dilute alkali. 3. The degradative studies described suggest that the poly(ribitol phosphate) chains of the wall teichoic acid may be attached to the wall by linkage to this glycerol phosphate oligomer.

Cell Wall↗

A novel teichoic acid from the cell wall of Streptomyces sp. VKM Ac-2275.

The cell wall of a pathogenic strain Streptomyces sp. VKM Ac-2275 isolated from potato tubers infected by scab contains a teichoic acid related to poly(glycosylpolyol phosphate) with a repeating unit established by chemical and NMR spectroscopic methods. About 60% of l-rhamnose residues bear an O-acetyl group at O-2 and 20% of the internal glucose residues contain an additional phosphate at C-4. The polymer is built of 5-6 units. This structure is found in bacteria for the first time. The strain is phylogenetically closest to the scab-causing species Streptomyces scabiei and Streptomyces europaeiscabiei, but differs from both these species in morphological and physiological characters and does not produce thaxtomin A, the main phytotoxin produced by S. scabiei.

Carbohydrate Sequence↗

Coordinated incorporation of nascent peptidoglycan and teichoic acid into pneumococcal cell walls and conservation of peptidoglycan during growth.

Choline-containing pneumococcal cell wals are sensitive to autolysin, whereas ethanolamine-containing walls are not. Bacteria were labeled with radioactive peptidoglycan precursors while growing either in choline- or in ethanolaminecontaining media. Subsequently, the labeled cells were allowed to grow for four to five generations in nonradioactive medium supplemented with the alternative amino alcohol source (i.e. cells labeled in choline medium yields ethanolamine; cells labeled in ethanolamine medium yields choline). The autolysin sensitivity of the isotope label in cell walls prepared from such bacteria indicates that nascent peptidoglycan and teichoic acid units that are synthesized at the same time are attached to one another, incorporated into the cell surface at the cellular equator, and remain conserved during growth the division of the bacteria.

Alanine↗

[Significance of the determination of teichoic acid antibodies in the diagnosis of Staphylococcus aureus bacteremia].

Sera of 47 patients with serious Staphylococcus aureus infections, 55 patients with septicemias caused by other organisms and 147 healthy controls were examined for teichoic acid antibodies by ELISA, counterimmunoelectrophoresis (CIE) and immunodiffusion (ID). The specificity and the predictive value of positive results for metastatic-septic complications were 100% each for ELISA, 93%/88% for CIE and 87%/80% for ID. The low sensitivity with all three test methods of 50-60% relates to single sera. The indication for prolonged antibiotic treatment should therefore be derived only from the analysis of consecutive serum samples. With 12% of the controls above the cut-off, the ID test seems not a very good screening test.

Anti-Bacterial Agents↗

The essential nature of teichoic acids in Bacillus subtilis as revealed by insertional mutagenesis.

A 30 kb DNA segment from the region of the Bacillus subtilis strain 168 chromosome which contains most, if not all, loci specifically involved in teichoic acid biosynthesis, has been cloned. A restriction map was established to which genetic markers were assigned. Four loci, tagA, tagB, gtaA and gtaD, are located on a DNA segment of about 7 kb, whereas the gtaB locus lies some 10 kb distant. The tagA and tagB loci are apparently transcribed independently. Insertional mutagenesis, using integrational plasmids carrying relevant fragments from the tag region, provides strong evidence that biosynthesis of polyglycerol phosphate [poly(groP)], so far largely considered as a dispensable polymer, is in fact essential for growth.

Bacillus subtilis↗

Correlation of teichoic acid D-alanyl esterification with the expression of methicillin resistance in Staphylococcus aureus.

Methicillin-susceptible and -resistant isolates of Staphylococcus aureus, including a teichoic acid-deficient mutant, were examined under varying physiological conditions for their degree of teichoic acid and lipoteichoic acid D-alanyl esterification. Methicillin-resistant strains grown in the presence of methicillin and NaCl possessed significantly decreased amounts of D-alanyl ester when compared with methicillin-susceptible isolates. These strains also exhibited reduced autolysis. An autoradiographic procedure was used to detect mutants, isolated by Tn917 and nitrosoguanidine mutagenesis of methicillin-susceptible strains, which were defective in D-alanyl ester formation. The quantitative uptake of D-[14C]alanine in these mutants was determined and the effect of methicillin on the growth and viability of each mutant was compared with the wild-type strain. S. aureus mutant strains, defective in the uptake and incorporation of D-alanine, were shown to exhibit slightly reduced autolysis and an enhanced expression of methicillin resistance.

Alanine↗

Analysis of teichoic acid biosynthesis regulation reveals that the extracytoplasmic function sigma factor sigmaM is induced by phosphate depletion in Bacillus subtilis W23.

The expression of the Bacillus subtilis W23 tar genes specifying the biosynthesis of the major wall teichoic acid, the poly(ribitol phosphate), was studied under phosphate limitation using lacZ reporter fusions. Three different regulation patterns can be deduced from these beta-galactosidase activity data: (i) tarD and tarL gene expression is downregulated under phosphate starvation; (ii) tarA and, to a minor extent, tarB expression after an initial decrease unexpectedly increases; and (iii) tarO is not influenced by phosphate concentration. To dissect the tarA regulatory pattern, its two promoters were analysed under phosphate limitation: The P(tarA)-ext promoter is repressed under phosphate starvation by the PhoPR two-component system, whereas, under the same conditions, the P(tarA)-int promoter is upregulated by the action of an extracytoplasmic function (ECF) sigma factor, sigma(M). In contrast to strain 168, sigma(M) is activated in strain W23 in phosphate-depleted conditions, a phenomenon indirectly dependent on PhoPR, the two-component regulatory system responsible for the adaptation to phosphate starvation. These results provide further evidence for the role of sigma(M) in cell-wall stress response, and suggest that impairment of cell-wall structure is the signal activating this ECF sigma factor.

Bacillus subtilis↗

Possible role of a choline-containing teichoic acid in the maintenance of normal cell shape and physiology in Streptococcus oralis.

Streptococcus oralis ATCC 35037 took up radioactively labeled choline from growth medium. Most of the choline (80 to 90%) was incorporated into the cell wall teichoic acid, and about 10% was localized in the plasma membrane. While cells grew in choline-free medium, they did so at slow rates and produced cell walls with greatly reduced amounts of phosphate and no detectable choline. Cells grown in choline-free medium had grossly abnormal shape and size. Both biochemical and morphological abnormalities were reversible by addition of choline to the medium.

Choline↗

Role of teichoic acids in Staphylococcus aureus nasal colonization, a major risk factor in nosocomial infections.

Colonization of the anterior nares in approximately 37% of the population is a major risk factor for severe Staphylococcus aureus infections. Here we show that wall teichoic acid (WTA), a surface-exposed staphylococcal polymer, is essential for nasal colonization and mediates interaction with human nasal epithelial cells. WTA-deficient mutants were impaired in their adherence to nasal cells, and were completely unable to colonize cotton rat nares. This study describes the first essential factor for S. aureus nasal colonization.

Animals↗

Comparison of anti-alpha-haemolysin and teichoic acid antibody tests in patients with Staphylococcus aureus endocarditis or bacteraemia.

To assess the value of serological tests in differentiating endocarditis and complicated bacteraemia due to Staphylococcus aureus from uncomplicated S. aureus bacteraemia and from nonstaphylococcal endocarditis we measured teichoic acid antibodies (TAA) (by means of a gel diffusion method) and antibodies to alpha-haemolysin (ASta) in the serum of 22 patients with S. aureus endocarditis, 42 patients with complicated S. aureus bacteraemia, 21 patients with uncomplicated S. aureus bacteraemia, 27 patients with other than S. aureus endocarditis, 17 patients with culture-negative endocarditis and 337 non-infected control patients. TAA and ASta titres were found significantly more often in staphylococcal endocarditis than in non-staphylococcal endocarditis, 59 per cent versus 4 per cent for the TAA test and 32 per cent versus 0 per cent for the ASta assay. The combined use of the two tests proved best for differentiating the two groups of endocarditis from each other, 72 per cent versus 4 per cent respectively. In the culture-negative endocarditis group there were two serologically positive patients whose anti-staphylococcal antibiotic therapy based on the serological findings was successful, supporting the clinical usefulness of staphylococcal serological tests in endocarditis of unknown bacterial aetiology. The serological tests were not useful, however, in differentiating S. aureus endocarditis from complicated or uncomplicated bacteraemia due to S. aureus without endocarditis.

Adolescent↗

A novel mannitol teichoic acid with side phosphate groups of Brevibacterium sp. VKM Ac-2118.

The cell wall of Brevibacterium sp. VKM Ac-2118 isolated from a frozen (mean annual temperature -12 degrees C) late Pliocene layer, 1.8-3 Myr, Kolyma lowland, Russia, contains mannitol teichoic acid with a previously unknown structure. This is 1,6-poly(mannitol phosphate) with the majority of the mannitol residues bearing side phosphate groups at O-4(3). The structure of the polymer was established by chemical methods, NMR spectroscopy, and MALDI-TOF mass spectrometry.

Brevibacterium↗

Cell walls, peptidoglycans, and teichoic acids of gram-positive bacteria as polyclonal inducers and immunomodulators of proliferative and lymphokine responses of human B and T lymphocytes.

In this study, the mitogenic and immunomodulating effects of bacterial cell wall preparations were investigated. Cell walls, peptidoglycans, and teichoic acids from Bacillus subtilis and Staphylococcus aureus Wood 46 activated both human T cells (supplemented with 10% monocytes) and B cells to proliferative and to produce leukocyte migration inhibitory factor. Similar results were obtained with adult and umbilical cord blood cells, suggesting that these bacterial preparations acted as mitogens. Cell walls and peptidoglycans had a modulating effect on purified protein derivative-induced and protein A-induced proliferation. In the presence of suboptimal concentrations of these stimulants, bacterial components enhanced the proliferative response. However, at optimal concentrations of purified protein derivative or protein A, bacterial components suppressed lymphocyte proliferation. Peptidoglycans solubilized by lysozyme activated B lymphocytes but not T cells. Solubilization had no effect on immunomodulating capacity.

Age Factors↗

Cell walls, peptidoglycans, and teichoic acids of Gram-positive bacteria as polyclonal inducers and immunomodulators of proliferative and lymphokine responses of human B and T lymphocytes.

In this study the mitogenic and immunomodulating effects of bacterial cell wall preparations were investigated. Cell walls, peptidoglycans, and teichoic acids from Bacillus subtilis and Staphylococcus aureus Wood 45 activated both human T cells (supplemented with 10% monocytes) and B cells to proliferate and to produce leukocyte-inhibitory factor. Similar results were obtained with adult and umbilical cord blood cells, suggesting that these bacterial preparations were acting as mitogens. Cell walls and peptidoglycans had a modulating effect on purified protein derivative- or protein A-induced proliferation. In the presence of suboptimal concentrations of these stimulants, bacterial components enhanced the proliferative response. However, at optimal concentrations of purified protein derivative or protein A, bacterial components suppressed lymphocyte proliferation. Peptidoglycans solubilized by lysozyme activated B lymphocytes but not T cells. Solubilization had no effect on the immunomodulating capacity.

B-Lymphocytes↗

Poly(glucosyl-N-acetylgalactosamine 1-phosphate), a wall teichoic acid of Bacillus subtilis 168: its biosynthetic pathway and mode of attachment to peptidoglycan.

The ggaAB operon of Bacillus subtilis 168 encodes enzymes responsible for the synthesis of poly(glucosyl N-acetylgalactosamine 1-phosphate) [poly(GlcGalNAc 1-P)], a wall teichoic acid (WTA). Analysis of the nucleotide sequence revealed that both GgaA and GgaB contained the motif characteristic of sugar transferases, while GgaB was most likely to be bifunctional, being endowed with an additional motif present in glucosyl/glycerophosphate transferases. Transcription of the operon was thermosensitive, and took place from an unusually distant sigma(A)-controlled promoter. The incorporation of the poly(GlcGalNAc 1-P) precursors by various mutants deficient in the synthesis of poly(glycerol phosphate), which is the most abundant WTA of strain 168, revealed that both WTAs were most likely to be attached to peptidoglycan (PG) through the same linkage unit (LU). The incorporation of poly(GlcGalNAc 1-P) precursors by protoplasts confirmed the existence of this LU, and provided further evidence that incorporation takes place at the outer surface of the protoplast membrane. The data presented here strengthen the view that biosynthesis of the LU, and the hooking of the LU-endowed polymer to PG, offer distinct widespread targets for antibiotics specific to Gram-positive bacteria.

Bacillus subtilis↗