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Biomedical subjects

I Ofek

Publications and source records attributed to I Ofek.

At least 91 records · Page 5Linked to original sources
PubMed1982

Differential effects of antibiotics on adhesins of antibiotic resistant strains of Escherichia coli.

Sublethal concentrations of antibiotics that inhibit protein synthesis were found to diminish the ability of strains of Escherichia coli to bind to human leukocytes and to oral epithelial cells, guinea pig erythrocytes, and mannan-containing yeast cells. In general depressed adherence correlated with diminished production of type 1 fimbriae by the drug-treated bacteria and was affected by the antibiotics in the following order: aminoglycoside (gentamicin, streptomycin, neomycin) greater than spectinomycin greater than tetracycline greater than chloramphenicol. With one notable exception, mutation to resistance to antibiotic-induced growth inhibition resulted in resistance to the sublethal, anti-adherence effects of the same antibiotic. The exceptional strain, VL-2, was the only one out of many streptomycin-resistant strains that was not also resistant to the anti-adherence effects of subinhibitory concentrations of the drug. Compared to control cultures of VL-2, those grown in antibiotic demonstrated decreased amounts of mannose-sensitive hemagglutination (by greater than 99%) and adherence to human epithelial cells (58%) and leukocytes (93%). When treated bacteria were examined by electron microscopy, they were found to be as heavily fimbriate as control bacteria, but their fimbriae were twice as long. Moreover, fimbriae isolated and purified from drug-treated bacteria had no lectin-like properties. Thus, although most antibiotics diminished the adhesive properties only the antibiotics-sensitive bacteria, streptomycin caused one streptomycin-resistant strain of bacteria to produce aberrant (non-adhesive) fimbrial protein.

Animals↗
PubMed1981

Sublethal concentrations of antibiotics and bacterial adhesion.

Various antibiotics in sublethal concentrations markedly impair adhesion of Streptococcus pyogenes and Escherichia coli to human cells. In streptococcal cells penicillin G caused an enhances loss of lipoteichoic acid, the ligand (adhesion) that binds the organism to host cells, with consequent loss of their adhesive properties. In E coli sublethal concentrations of penicillin prevented the surface expression of the mannose-specific adhesion by distorting cell wall biosynthesis. In contrast to streptococci, E coli cells could not be made to lose their adhesions once their adhesions once they had been formed. Streptomycin in subinhibitory concentration similarly suppressed the acquisition of mannose-binding and adhesive activities in several strains of antibiotic-sensitive E. coli but not in isogenic derivatives with ribosomal mutation to high-level streptomycin resistance, rpsL, or in bacteria in the stationary phase of growth, suggesting that streptomycin exerted its sublethal suppressive effects by classic mechanisms of action on the bacterial ribosome. Strain VL2, derived from one streptomycin-resistant mutant, retained a high level (1000 microgram/ml) of resistance to streptomycin but reacquired sensitivity to the sublethal effect; growth in 30 microgram streptomycin/ml suppressed mannose-sensitive haemagglutination (less than 1% of control) as well as mannose-sensitive adhesion to epithelial cells (42%) or leucocytes (7%). Although these streptomycin-treated bacteria demonstrated an unaltered degree of fimbriation their fimbriae were significantly longer than those on the untreated bacteria. Furthermore, in contrast to the untreated bacteria, the fimbriae isolated from the drug-treated bacteria were found to lack mannose-binding activity as measured by haemagglutination. It therefore, appears that streptomycin can cause even resistant bacteria to produce an aberrant fimbrial protein, presumably by causing misreading in "competent" ribosomes. These studies indicate that the use of sublethal doses of certain antibiotics whose mode of action is well known may shed light on the genetic and chemical modulation of bacterial factors involved in mucosal colonization.

Adhesiveness↗
PubMed1981

Bacterial adherence to cell surface sugars.

Bacterial adherence to animal cell surfaces is of interest because of its relation to pathogenicity and the insight it provides into determinants of intercellular recognition. The attachment of various strains of Escherichia coli and Salmonella spp. to epithelial cells and phagocytes is inhibited by D-mannose, and the adherence of other bacteria is inhibited by sugars such as L-fucose and D-galactose, suggesting that sugar-mediated adherence is widespread. This intercellular recognition is thought to be mediated by sugar residues (e.g. D-mannose) on the surface of animal cells, to which bacteria attach by a sugar-binding substance on their surface. The nature of the receptors on the animal cells is unknown. There is evidence that E. coli produces lectin-like substances specific for D-mannose, by which it binds to the cells. The most common form of these lectin-like substances appears to be the bacterial pili, which can be reversibly dissociated into their protein subunits. The lectin can also be in the form of bacterial flagella or tightly attached to the outer membrane of the bacteria. Mannose-specific attachment may assist bacteria in colonizing and invading their hosts: methyl alpha-D-mannoside (but not methyl alpha-D-glucoside) significantly reduced infection of the urinary tract of mice by virulent strains of E. coli. Once bacteria penetrate the host their ability to binding sugars on phagocytes may impair their virulence by facilitating phagocytosis. Further studies of the sugar-mediated bacterial adherence by organisms growing in vivo and the structural identification of the host cell receptors may lead to the design of more effective adherence inhibitors that may help to prevent certain bacterial infections.

Adhesiveness↗
PubMed1981

Loss of lectin-like activity in aberrant type 1 fimbriae of Escherichia coli.

Growth of a streptomycin-resistant strain of Escherichia coli (VL-2) in the presence of 30 microgram of streptomycin per ml resulted in the production by these bacteria of structurally altered, nonfunctional type 1 fimbriae. This strain, when grown in this subinhibitory concentration of streptomycin, became incapable of producing mannose-sensitive hemagglutination (<1% of that of the control). Adhering ability to epithelial cells and human leukocytes was also diminished (42 and 7% of that of the control, respectively). Although these streptomycin-treated bacteria were as heavily fimbriated as untreated bacteria, their fimbriae were significantly longer. Furthermore, in contrast to the fimbriae of the untreated bacteria, those isolated from the drug-treated bacteria were found to lack mannose binding activity as measured by hemagglutination. It appears, therefore, that streptomycin can cause even resistant bacteria to produce an aberrant fimbrial protein, possibly by causing misreading of messenger RNA. These studies indicate that the use of sublethal doses of certain antibiotics whose mode of action is well known may shed light on the genetic and chemical modulation of bacterial factors involved in mucosal colonization.

Drug Resistance, Microbial↗
PubMed1981

Characterization of lipoteichoic acid binding to polymorphonuclear leukocytes of human blood.

Human polymorphonuclear leukocytes (PMN) were shown to possess specific binding sites for lipoteichoic acid (LTA). LTA binding was reversible and time and temperature dependent. Scatchard plot analysis revealed an apparently single population of 6.6 X 10(6) LTA binding sites per PMN with a dissociation constant of 5.6 microM. Attachment of an avirulent, unencapsulated, M-negative strain of group A streptococci to PMN was inhibited by LTA, but not by other bacterial somatic antigens tested. Occupation of 30% of the LTA binding sites resulted in greater than 70% inhibition of streptococcal attachment to PMN. In contrast, LTA failed to block attachment of Escherichia coli or antibody-coated streptococci, indicating that binding sites for E. coli and the Fc portion of immunoglobulin G are distinct from those for LTA. Immunofluorescent studies demonstrated that LTA remained uniformly bound to PMN membranes for as long as 2 h at 37 degrees C. Cross-linking of PMN-bound LTA with anti-LTA resulted in rapid capping of LTA receptor sites. The results suggest that LTA is a monovalent ligand interacting with mobile receptors in the plasma membrane of PMN.

Binding Sites↗
PubMed1981

Adherence of Acinetobacter calcoaceticus RAG-1 to human epithelial cells and to hexadecane.

The ability of Acinetobacter calcoaceticus RAG-1 to adhere to human epithelial cells was investigated and compared with its ability to adhere to a test hydrocarbon (hexadecane). RAG-1, a microorganism originally isolated for growth on hydrocarbon, adhered to epithelial cells when grown under conditions which promote its adherence to hexadecane; similarly, RAG-1 cells adhered poorly to epithelial cells when grown under conditions which cause the cells to possess low affinity towards hexadecane. A mutant derived from RAG-1, MR-481, deficient in its ability to adhere to hydrocarbon, was similarly unable to adhere to epithelial cells. RAG-1 adherence to epithelial cells was not blocked by a number of sugars tested. Streptococcus pyogenes, whose adherence to epithelial cells has been previously attributed to hydrophobic interactions, was also able to adhere to hexadecane. Results suggest that hydrophobic interactions mediate adherence of the strains studied to both epithelial cells and hydrocarbon.

Acinetobacter↗
PubMed1981

Mannose-specific adherence of Escherichia coli freshly excreted in the urine of patients with urinary tract infections, and of isolates subcultured from the infected urine.

The mannose-specific adherence to yeast cells of Escherichia coli excreted in the urine of patients with urinary tract infections was compared with that of isolates from the same urine after growth of the bacteria in broth. The results revealed that although E. coli excreted in only 2 of 24 urine specimens exhibited mannose-specific adherence, about half of the broth cultures from these specimens did so. Examination of representative specimens of E. coli excreted in urine showed that coating antibodies, mannose-containing glycoproteins, and encapsulation were not responsible for the lack of the mannose-specific adherence. Our results suggest that E. coli strains that are genetically capable of exhibiting mannose-specific adherence may, when growing in the bladder, be in a phase of growth which suppresses the phenotypic expression of this trait. Mannose-specific adherence is indicative of the presence on the bacterial surface of adhesions (lectins) that bind the organisms to mannose residues on both epithelial and phagocytic cells. We propose that whereas at the initial stages of infection the bacteria may benefit from their ability to bind to mannose residues on epithelial cells, loss of this ability at the later stages of the infection is also beneficial, since the bacteria can no longer adhere to mannose residues on phagocytes, and are thus resistant to nonimmune phagocytosis.

Cell Adhesion↗
PubMed1980

Binding of streptococcal lipoteichoic acid to the fatty acid binding sites on serum albumin.

The ability of the fatty acid binding sites of serum albumin to bind lipoteichoic acid of Streptococcus pyogenes was investigated. Initial studies indicated that lipoteichoic acid, but not its deacylated deprivative, protected albumin from being denatured by heat (80C for 1 h) and changed its mobility in an electrical field. Albumin covalently linked to agarose beads bound radiolabeled lipoteichoic acid, and the bound [3H]lipoteichoic acid could be specifically eluted with unlabeled lipoteichoic acid or albumin but not with other proteins tested. After binding to albumins, the lipoteichoic acid also could be quantitatively eluted with 50% ethanol and various detergents but not with up to 1.0 M sodium chloride. Binding of lipoteichoic acid to albumin followed first order kinetics, reaching saturation at 12 h. Analysis of the binding data by a Scatchard plot indicated heterogeneity of the binding sites on the albumin molecule similar to that previously reported for fatty acids. The affinity of binding of lipoteichoic acid to albumin was found to be intermediate between that previously reported for octanoic and palmitic acids, respectively. Based on these findings, we prepared affinity columns of immobilized albumin and were able to separate biologically active lipoteichoic acid from heterogeneous extracts of S. pyogenes.

Binding Sites↗
PubMed1980

Heterogeneity of type-specific and cross-reactive antigenic determinants within a single M protein of group A streptococci.

The heterogeneity of a pepsin extract of type-24 M protein (pep M24) was demonstrated by absorption of type-specific and cross-reactive human antisera with M protein fragments and heterologous serotypes of M proteins, pepsin extract of type-5 M protein (pep M5) and pepsin extract of type-6 M protein (pep M6). 2 of 12 individuals immunized with pep M24 developed significant rises in antibody titers against pep M5 and pep M6, as measured by the enzyme-linked immunosorbent assay. The sam individuals also developed opsonic antibodies against type-6, but not type-5, streptococci, which suggested the development of cross-protective immunity. Inhibition studies of one of these sera with the heterologous pep M proteins showed that the cross-reactive antibodies against pep M6 could not be blocked by high concentrations of pep M24, the immunizing antigen; these antibodies could be blocked, however, by cyanogen bromide-derived peptide fragments of pep M24, which suggested that the cross-reactive antibody was raised against an inaccessible site(s) in the pep M24 molecule. Inhibition studies of type-specific immune sera with pep M24 and peptides derived therefrom indicated that the M protein molecule contained multiple distinct as well as identical type-specific antigenic determinants that are unequally distributed among the seven derived peptide fragments.

Bacterial Proteins↗
PubMed1980

Characteristics of the binding of streptococcal lipoteichoic acid to human oral epithelial cells.

Membrane receptors for lipoteichoic acid (LTA) may be involved in the adherence of streptococci to human pharyngeal epithelial cells. Since streptococcal binding to oral epithelial cells has been shown to be low at birth with a gradual increase to adult levels within three days, the characteristics of the binding of radiolabeled LTA to epithelial cells were investigated. LTA binding to epithelial cells from both infants and adults was time- and cell concentration-dependent. Binding was reversible in the presence of a 50-fold excess of unlabeled LTA. The number of LTA-binding sites in infant cells was only one-half that of adult cells; the amount of LTA bound increased to adult levels within three days of birth and paralleled an increase in the ability of these cells to bind streptococci. Thus, oral epithelial cells may possess specific binding sites for LTA, and a critical number or arrangement of LTA-binding sites may be required for the optimal binding of streptococci.

Adult↗
PubMed1980

Influence of sublethal concentrations of antibiotics on the expression of the mannose-specific ligand of Escherichia coli.

The ability of streptomycin, in subinhibitory concentrations, to differentially suppress the acquisition of the mannose-binding activity of Escherichia coli was demonstrated in several strains, but not one with a ribosomal mutation to high-level streptomycin resistance, rpsL. We also determined that the growth of bacteria in other antibiotics, notably those that interfere with protein synthesis, resulted in diminished mannose-binding activity (as measured by yeast cell agglutination), degree of piliation (as measured by electron microscopy), and adherence to human oral epithelial cells. The aminoglycoside antibiotics streptomycin, gentamicin, and neomycin had the most marked effects relative to their minimum inhibitory concentrations, followed by tetracycline. Both spectinomycin and chloramphenicol had more effect on adherence than on piliation, although spectinomycin had a more pronounced effect on mannose-binding activity than did chloramphenicol. We conclude that antibiotics, at concentrations below their minimum inhibitory concentration, may have profound effects on surface properties of bacteria that may be pertinent for their ability to colonize and infect human mucosal surfaces. The mechanism(s) may vary from one drug to another, but appear to depend on the classic actions of the antibiotics on inhibiting protein synthesis.

Anti-Bacterial Agents↗
PubMed1980

Fatty acid binding sites of serum albumin as membrane receptor analogs for streptococcal lipoteichoic acid.

The ability of bovine serum albumin to inhibit the binding of group A streptococcal lipoteichoic acid (LTA) to human cells was investigated. Albumin blocked the ability of LTA to sensitize erythrocytes to agglutinate in the presence of anti-LTA in a dose-dependent manner. The inhibition of LTA binding to erythrocytes was demonstrated directly with radiolabeled LTA. At an albumin/LTA molar ratio of 1.5:1, albumin binding of the radiolabeled LTA at erythrocytes was inhibited by 45%. Analysis of the binding of radiolabeled LTA to erythrocytes in the presence of albumin indicated that albumin competitively inhibited the binding of LTA to putative cell membrane receptors, indicating that albumin and erythrocytes both bind to the same moiety on the LTA molecule.

Binding Sites↗
PubMed1980

Mannose-binding activity of Escherichia coli: a determinant of attachment and ingestion of the bacteria by macrophages.

Recently, it was suggested that a mannose-specific lectin on the bacterial cell surface is responsible for the recognition by phagocytic cells of certain nonopsonized Escherichia coli strains. In this study we assessed the interaction of two strains of E. coli at different phases of growth with a monolayer of mouse peritoneal macrophages and developed a direct method with [(14)C]mannan to quantitate the bacterial mannose-binding activity. Normal-sized bacteria were obtained from logarithmic and stationary phases of growth. Nonseptated filamentous cells were formed by growing the organisms in the presence of cephalexin or at a restrictive temperature. Attachment to macrophages of all bacterial forms was inhibited by methyl alpha-d-mannoside and mannan but not by other sugars tested. The attachment of stationary phase and filamentous bacteria to macrophages, as well as their mannose-binding activity, was similar, whereas in the exponential-phase bacteria they were markedly reduced. The results show a linear relation between the two parameters (R = 0.98, P < 0.001). The internalization of the filamentous cells attached to macrophages during 45 min of incubation was much less efficient (20%) compared to that of exponential-phase, stationary-phase, or antibody-coated filamentous bacteria (90%). The results indicate that the mannose-binding activity of E. coli determines the recognition of the organisms by phagocytes. They further suggest that administration of beta-lactam antibiotics may impair elimination of certain pathogenic bacteria by inducing the formation of filaments which are inefficiently internalized by the host's phagocytic cells.

Animals↗
PubMed1980

Lipoteichoic acid-binding and biological properties of T protein of group A streptococcus.

T protein was extracted with trypsin from an avirulent, M protein-deficient, type 1 group A Streptococcus and purified by ammonium sulfate precipitation and anion-exchange chromatography. The latter procedure removed contaminating lipoteichoic acid (LTA) from the T protein, which consisted of a heterogeneous mixture of polypeptides resistant to digestion by trypsin and ranged in molecular size from 160,000 to 200,000 daltons. Threonine, aspartic acid, glutamic acid, lysine, and valine were the most predominant amino acids. The binding of LTA to an affinity column of T protein was reversible with increasing concentrations of ethanol but not with increasing ionic strength. T protein bound less palmitic acid and LTA than did fatty acid-free bovine albumin and did not stimulate human peripheral lymphocytes. Because the surface and cell wall distribution of the T proteins and LTA appear similar, the possibility exists that T proteins and LTA may interact in situ by weakly hydrophobic bonds. Such ligand-ligand interaction may be indirectly involved in the adherence of group A streptococci to host cell membranes that is known to be mediated by LTA.

Amino Acids↗
PubMed1979

Human immune response to immunization with a structurally defined polypeptide fragment of streptococcal M protein.

We tested the ability of pepsin-extracted, highly purified M protein to induce type-specific immunity in experimental animals and humans. M protein was prepared from limited peptic digests of whole group A type 24 streptococci and was purified to chemical homogeneity as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, quantitative amino acid analysis, and Edman degradation. For vaccination, the lyophilized M24 protein preparation (pep M24) was precipitated in aluminum hydroxide. When injected into laboratory animals, alum-precipitated pep M24 produced type-specific protective antibodies and was free of non-type-specific immunoreactivity. In man, skin tests with 1-microgram doses of pep M24 were negative in all 37 adults tested. 12 adult human volunteers received two-four subcutaneous injections of 100-200 micrograms of alum-precipitated pep M24 at intervals of at least 2 wk. The immune response to pep M24 was measured by a variety of assays designed to detect (a) type-specific humoral antibodies (opsonophagocytic, long chain, and mouse protection tests); (b) total humoral antibodies (complement fixation and enzyme-linked immunosorbent assay); (c) cellular immunity (skin tests); and (d) heart cross-reactive antibodies (immunofluorescence). Type-specific opsonic antibodies developed in 10 of the 12 vaccinees, and positive delayed-type skin tests developed in 11. Immune sera from two of the vaccinees were effective in mouse-protection tests against challenge with M24 but not M6 streptococci. None of the volunteers developed heart-reactive antibodies or antibodies to non-type-specific M protein antigens. Alum-precipitated pep M24 was well-tolerated in man, and no serious local or systemic reactions were observed. Thus, pep M24 induces type-specific, protective antibodies in doses that are well-tolerated in man.

Adult↗