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Biomedical subjects

I Ofek

Publications and source records attributed to I Ofek.

At least 37 records · Page 2Linked to original sources
PubMed1991

Lectinophagocytosis of encapsulated Klebsiella pneumoniae mediated by surface lectins of guinea pig alveolar macrophages and human monocyte-derived macrophages.

Macrophages express a mannose/N-acetylglucosamine-specific lectin which serves as a receptor for nonopsonic phagocytosis of mannose-coated particles. We have examined the binding to guinea pig alveolar macrophages in a serum-free medium of 16 Klebsiella pneumoniae serotypes and of the capsular polysaccharides isolated from 7 of these serotypes. Only five polysaccharides containing the repeating sequence Man alpha 2/3Man or L-Rha alpha 2/3-L-Rha bound to the macrophages. Of the 11 bacterial serotypes expressing such disaccharides in their capsular polysaccharides, 7 bound efficiently, 2 bound poorly, and 2 did not bind at all. No binding occurred with five serotypes lacking these disaccharides. Binding of the bacteria was inhibited by homologous and heterologous capsular polysaccharides that contain the disaccharide sequences, by mannan, and by (Man)25BSA (where BSA is bovine serum albumin). Man alpha 2/3Man-containing oligosaccharides were potent inhibitors compared with monosaccharides. Binding was dependent on Ca2+, modulated by cultivating the macrophages on mannan-coated surfaces, and increased in human monocyte-derived macrophages compared with monocytes. The bulk of the bacteria bound to the macrophages was internalized and killed. The data taken together suggest that Klebsiella pneumoniae cells undergo lectinophagocytosis mediated by capsular disaccharides recognized by the mannose/N-acetylglucosamine-specific lectin of macrophages. This may enhance clearance of the organisms from the serum-poor environment of the lung.

Animals↗
PubMed1991

Prospective study of the association between serum antibodies to lipopolysaccharide O antigen and the attack rate of shigellosis.

A means for determining immune status against shigellosis would significantly improve the design and evaluation of interventional and other epidemiologic studies. Previous case-control studies have indicated the potential role of humoral antilipopolysaccharide antibodies. To test this proposition, 190 soldiers serving in a field unit were monitored prospectively for 2.5 months for shigellosis. Blood samples were taken at the beginning of the follow-up period and tested for serological evidence of prior exposure to Shigella sonnei and Shigella flexneri. The risk for acquiring S. sonnei shigellosis was 3.7 times higher for individuals lacking homologous antibodies (P less than 0.02). The risk for acquiring S. flexneri shigellosis was 2.4 times higher for individuals lacking antibodies, although a low attack rate for S. flexneri resulted in numbers too small to achieve statistical significance. While the importance of the serum antilipopolysaccharide antibodies in protection against the disease remains unclear, these findings demonstrate that they are strong markers of acquired immunity. Serological markers should be incorporated in epidemiologic studies of shigellosis and in the design and evaluation of future trials of potential anti-Shigella vaccines.

Antibodies, Bacterial↗
PubMed1990

Inability of toxin inhibitors to neutralize enhanced toxicity caused by bacteria adherent to tissue culture cells.

Toxicity to Y-1 adrenal mouse cells caused by heat-labile toxin secreted by an enterotoxigenic strain of Escherichia coli (H-10407-p) was 40-fold enhanced in mixtures containing organisms capable of adhering to the Y-1 cells compared with monolayers exposed to organisms whose adherence was inhibited by mannoside. Severalfold the concentrations of anti-heat-labile toxin antibodies required to neutralize the toxicity of nonadherent bacteria were unable to neutralize the toxicity caused by adherent bacteria. The cytolytic activity toward tissue culture cells and mouse peritoneal macrophages caused by streptolysin S carried by Streptococcus pyogenes was severalfold increased in mixtures containing organisms capable of adhering to the target cells compared with mixtures containing nonadherent bacteria. The ability of trypan blue and RNA core to inhibit the cell-bound streptolysin S was determined in tissue culture cells containing adherent streptococci and mixtures of streptococci randomly colliding with erythrocytes. Both inhibitors were markedly less effective in neutralizing cytolysis than in their ability to neutralize hemolysis. We conclude that compared with toxins produced by nonadherent bacteria, those produced by bacteria adherent to cells are targeted more efficiently and become relatively inaccessible to neutralization by toxin inhibitors.

Animals↗
PubMed1990

Stimulation of oxidative burst in human monocytes by lipoteichoic acids.

Lipoteichoic acid isolated from Streptococcus faecalis or Streptococcus pyogenes caused direct activation of the respiratory burst in human peripheral blood monocytes. This activity appears to be related to the ability of lipoteichoic acid to bind to the monocyte membrane and trigger the polarization of receptors (capping).

Adult↗
PubMed1989

Lectinophagocytosis mediated by bacterial surface lectins.

The evidence showing that non-opsonic recognition of bacteria by phagocytes involve interaction between bacterial surface lectin and sugars on the phagocytic cells is summarized. This process, termed lectinophagocytosis, probably occur in vivo as evident from experimental infections with mixed phenotypes one of which express mannose specific (MS) lectin which mediate lectinophagocytosis of the bacteria and the other does not. In all cases studied, the lectin bearing phenotype survived better in phagocytes-poor sites and the phenotype which does not express this lectin survived better in phagocytes-rich sites. Due to the phase variation phenomenon, an off-on switch allowing the bacterial clone to alternate between lectin expressing and non-expressing phenotypes, the invading bacteria grow as a mixture of phenotypes. The phenotype expressing fimbrial lectin for which receptors are accessible on phagocytic cells undergo lectinophagocytosis. The phenotypes not expressing fimbrial lectin or expressing lectin for which receptors are not available on phagocytic cells may escape phagocytosis and proliferate. It is postulated that pathogenesis of inflammation and tissue damage following infections with MS lectin bearing bacteria may be partly due to both bacterial proliferation resulting in the release of toxic products and to lectinophagocytosis associated with the release of inflammatory agents.

Animals↗
PubMed1989

Inhibitory activity of cranberry juice on adherence of type 1 and type P fimbriated Escherichia coli to eucaryotic cells.

Inhibition of bacterial adherence to bladder cells has been assumed to account for the beneficial action ascribed to cranberry juice and cranberry juice cocktail in the prevention of urinary tract infections (A. E. Sobota, J. Urol. 131:1013-1016, 1984). We have examined the effect of the cocktail and juice on the adherence of Escherichia coli expressing surface lectins of defined sugar specificity to yeasts, tissue culture cells, erythrocytes, and mouse peritoneal macrophages. Cranberry juice cocktail inhibited the adherence of urinary isolates expressing type 1 fimbriae (mannose specific) and P fimbriae [specific for alpha-D-Gal(1----4)-beta-D-Gal] but had no effect on a diarrheal isolate expressing a CFA/I adhesin. The cocktail also inhibited yeast agglutination by purified type 1 fimbriae. The inhibitory activity for type 1 fimbriated E. coli was dialyzable and could be ascribed to the fructose present in the cocktail; this sugar was about 1/10 as active as methyl alpha-D-mannoside in inhibiting the adherence of type 1 fimbriated bacteria. The inhibitory activity for the P fimbriated bacteria was nondialyzable and was detected only after preincubation of the bacteria with the cocktail. Cranberry juice, orange juice, and pineapple juice also inhibited adherence of type 1 fimbriated E. coli, most likely because of their fructose content. However, the two latter juices did not inhibit the P fimbriated bacteria. We conclude that cranberry juice contains at least two inhibitors of lectin-mediated adherence of uropathogens to eucaryotic cells. Further studies are required to establish whether these inhibitors play a role in vivo.

Agglutination↗
PubMed1989

Immunoglobulin M, A, and G antibody response to lipopolysaccharide O antigen in symptomatic and asymptomatic Shigella infections.

The antilipopolysaccharide antibody response in sera obtained from subjects involved in 10 outbreaks of shigellosis occurring in Israeli military field units was determined by an enzyme-linked immunosorbent assay and a passive hemagglutination test. Both tests were found to be sensitive and specific for the diagnosis of shigellosis. A significant antibody response was detected in 73 to 82% of the symptomatic and 48 to 60% of the asymptomatic subjects during the Shigella sonnei and Shigella flexneri outbreaks. Fifty percent of the symptomatic and none of the asymptomatic subjects showed a significant antibody response in the Shigella boydii outbreaks. An examination of the kinetics of the antibody levels over a 10-week period after the onset of disease revealed that immunoglobulin A (IgA) levels were highest 2 weeks after infection and had declined to initial levels within 2.5 months. In contrast, IgG levels at the late convalescent stage were half those measured at early convalescence, still being about twice as high as the initial titers. Although the IgM levels showed a pattern similar to that of IgA, their elevation at the early convalescent stage was less pronounced. We conclude that the detection of an increase in the level of the IgA fraction appeared to be the best indicator for recent symptomatic, as well as symptomatic, infections due to Shigella organisms.

Adult↗
PubMed1988

Enzyme-linked immunosorbent assay for quantitation of attachment and ingestion stages of bacterial phagocytosis.

Research on phagocytosis of bacteria is often hampered by the inability to distinguish quantitatively between bacteria that have been ingested by phagocytic cells and those which are attached to the surface of the cells. A method using the enzyme-linked immunosorbent assay technique to simply and accurately measure the rate of bacterial ingestion by phagocytic cells is described. The method is based on the ability of antibacterial antibodies to bind to bacteria attached to but not internalized by phagocytic cells. The attached bacteria were quantitated by enzyme-linked immunosorbent assay. Compared with the number of bacteria at zero time (17 bacteria attached per phagocyte) only 10 to 20% of the bacteria remained attached to phagocytic cells after incubation for 30 min at 37 degrees C. The decrease in detected attached bacteria at 37 degrees C was due to internalization of the bacteria by phagocytic cells, since upon disruption of the monolayer, most of the ingested bacteria were recovered, and at 4 degrees C, most of the bacteria remained extracellularly attached. The proposed attachment and ingestion assay is easy to perform, allows the detection of specific attachment of test bacteria, and provides objective quantitation of attached and ingested bacteria. Most importantly, the assay allows testing of ingestion rates of bacteria under many variables on the same day.

Animals↗
PubMed1987

Hybridoma antibodies to the lipid-binding site(s) in the amino-terminal region of fibronectin inhibits binding of streptococcal lipoteichoic acid.

In this report, we present evidence to suggest that streptococci and lipoteichoic acid (LTA) interact with a fatty acid binding site located near the NH2-terminus of fibronectin. The evidence is based on the following observations. Antibodies directed against a synthetic peptide (residues 1-30 of the amino-terminus of fibronectin) reacted with the two thermolysin-generated peptides (24 and 28 kilodaltons [kDa]) that were adsorbed by and eluted from streptococci. The adsorption of the 24- and 28-kDa peptides to streptococci was inhibited by LTA. The two monoclonal antibodies that inhibited the binding of LTA to fibronectin reacted only with the 24- and 28-kDa fragments of fibronectin. Conversely, LTA, as well as lauric acid and oleic acid, blocked the binding of the same monoclonal antibodies to fibronectin. LTA had no effect on the binding of hybridoma antibodies directed against the collagen or cell-binding domain.

Animals↗
PubMed1987

Aromatic alpha-glycosides of mannose are powerful inhibitors of the adherence of type 1 fimbriated Escherichia coli to yeast and intestinal epithelial cells.

Adherence of bacteria via their surface lectins to host epithelial cells is considered an important initial event in bacterial pathogenesis. Mannose-specific (type 1) fimbriae are among the most commonly found lectins in enterobacteria. We studied the effect of aromatic alpha-glycosides of mannose on the agglutination of mannan-containing yeasts by different strains of Escherichia coli and on the adherence of the bacteria to guinea pig ileal epithelial cells. In both systems these compounds were considerably more effective inhibitors than methyl alpha-mannoside, with 4-methylumbelliferyl alpha-mannoside and p-nitro-o-chlorophenyl alpha-mannoside being the strongest inhibitors. Both compounds were approximately 400-times stronger inhibitors of yeast agglutination by E. coli O128 than was methyl alpha-mannoside and 1,000- and 470-fold stronger, respectively, than was methyl alpha-mannoside in inhibiting the adherence of the bacteria to ileal epithelial cells. 4-Methylumbelliferyl alpha-mannoside was 540 to 1,000 times more effective in inhibiting yeast agglutination by four additional strains of mannose-specific E. coli. It was also more efficient than methyl alpha-mannoside in removing adherent E. coli O128 from ileal epithelial cells. Our results provide further evidence that type 1 fimbriae of E. coli possess a hydrophobic region next to the mannose-binding site. The results suggest that 4-methylumbelliferyl alpha-mannoside and p-nitro-o-chlorophenyl alpha-mannoside are good candidates for the design of therapeutic agents that may prevent adherence in vivo and infection by E. coli strains that express type 1 fimbriae.

Agglutination↗
PubMed1987

Membrane glycoproteins of human polymorphonuclear leukocytes that act as receptors for mannose-specific Escherichia coli.

Type 1 fimbriated (mannose-specific) Escherichia coli cells bind to mannose residues on human polymorphonuclear leukocytes (PMN); this leads to phagocytosis of the bacteria. To identify the mannose-containing receptors on the PMN, the cells were surface labeled with 125I and lysed in 0.5% Nonidet P-40, and the lysate was fractionated by affinity chromatography on a column of Sepharose-bound fimbriae. Analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography of the material eluted from the column with 500 mM methyl-alpha-mannoside revealed two radioactive bands of Mr 70,000 to 80,000 (gp70-80) and 100,000 (gp100). Another weak band of Mr 150,000 (gp150) was observed after prolonged exposure of the gel. Upon blotting of the glycoproteins separated by polyacrylamide gel electrophoresis and overlaying of the blots with concanavalin A, gp150 appeared as the major band. Membrane preparations of the PMN were enriched in gp70-80, gp100, and gp150, in comparison with the cell homogenates, further suggesting that these glycoproteins are surface components. Fractionation of the membrane preparations on the immobilized fimbriae followed by concanavalin A overlay of blots of the methyl-alpha-mannoside-eluted material revealed that gp150 was the major component in this fraction. The eluted fraction, obtained from a cell lysate (4.4 micrograms/ml), inhibited by 70% the agglutination of yeasts by the intact bacteria. Our results suggest that the three surface glycoproteins isolated by us serve as receptors for mannose-specific E. coli on PMN and may be involved in the lectin-mediated phagocytosis of the bacteria.

Bacterial Adhesion↗
PubMed1987

Growth advantage and enhanced toxicity of Escherichia coli adherent to tissue culture cells due to restricted diffusion of products secreted by the cells.

This study was undertaken to examine whether Escherichia coli adherent to tissue cells gain advantages over nonadherent bacteria due to their proximity to the cells. We used tissue culture cells and isogenic derivatives of a proline auxotrophic strain of E. coli that were fimbriated (Fim+) or nonfimbriated (Fim-), and were heat-labile enterotoxin producing (Tox+) or toxin nonproducing (Tox-). We found that the Fim+ bacteria; which were capable of adhering to tissue culture cells, initiated growth much sooner than did nonadherent Fim- bacteria; the adherent bacteria used tissue cell-derived proline, which was available at high concentrations only in the zone of bacterial adherence. Likewise, cyclic AMP secreted by adherent (Fim+) bacteria was maintained at high concentration on the tissue cell surfaces. As few as 2 X 10(5) adherent Fim+ Tox+ bacteria exert toxic activity upon Y1 adrenal cells, whereas toxin secreted in the medium by 6 X 10(6) Fim- Tox+ bacteria was undetectable. The results suggest that the growth advantage and enhanced toxicity of adherent E. coli is due to restricted diffusion of products secreted by the tissue culture and bacterial cells, respectively.

Adrenal Glands↗
PubMed1986

Infant mouse model of adherence and colonization of intestinal tissues by enterotoxigenic strains of Escherichia coli isolated from humans.

The ability of enterotoxigenic Escherichia coli H10407, which possesses colonization factor antigen I, to colonize the intestinal mucosa of infant mice was considerably better than that of its colonization factor antigen I-negative derivative H10407-P. The latter strain previously was shown to lack cell adhering ability in vitro and to have a diminished capacity to infect human volunteers as compared with the parent strain. D-Mannose blocked both colonization by an enterotoxigenic E. coli isolate (801) possessing both mannose-resistant and mannose-sensitive adhesins and the in vitro adherence of the strain to intestinal segments of infant mice. A derivative of another enterotoxigenic E. coli strain (lacking both mannose-sensitive and mannose-resistant adhesins obtained by in vivo passage showed a significant increase in colonizing ability in comparison with the parent strain. We conclude that the infant mouse model of infection of intestinal mucosa complemented by in vitro adherence assays with excised intestinal tissue is suitable for the study of the bacterial properties responsible for the various stages of intestinal colonization by human enterotoxigenic E. coli.

Adhesiveness↗
PubMed1986

Binding of Streptococcus pyogenes to soluble and insoluble fibronectin.

The interaction of soluble and insoluble fibronectin with Streptococcus pyogenes was investigated. Soluble fibronectin bound to S. pyogenes in a dose-dependent and irreversible manner. Lipoteichoic acid competitively inhibited the binding of fibronectin to S. pyogenes but had little effect on the binding of fibronectin to staphylococci or pneumococci. The phase of growth of the streptococci had a slight effect on binding of fibronectin, with optimal binding occurring in the late log phase. S. pyogenes cells bound to fibronectin immobilized on microtiter plates in a dose-dependent and saturable manner. Both soluble fibronectin and lipoteichoic acid inhibited the binding of streptococci to immobilized fibronectin, suggesting that streptococci interact with soluble and insoluble fibronectin in a similar manner. Antibodies to fibronectin blocked the attachment of streptococci to immobilized fibronectin, whereas normal serum had no effect. Adherence of streptococci to buccal epithelial cells was inhibited by antibodies to fibronectin, but not by normal sera or by antibodies to buccal epithelial cells. The data suggest that lipoteichoic acid on the surface of S. pyogenes binds to fibronectin exposed on the host cell and that such binding mediates the attachment of streptococci to host cells.

Adhesiveness↗
PubMed1986

Enzyme-linked immunosorbent assay for adherence of bacteria to animal cells.

Epithelial cells scraped from human oral mucosa and from pig intestines were immobilized onto the flat bottom surfaces of microtiter plates to study the adherence of various bacterial species to host cells. Bacterial adherence was quantitated either by an enzyme-linked immunosorbent assay technique with specific antibacterial serum as the first antibody followed by peroxidase-conjugated second antibody or by using biotinylated bacteria and avidin-peroxidase as the detecting agent. Unlabeled Escherichia coli and purified E. coli 987P fimbriae inhibited the adherence of biotinylated E. coli to immobilized enterocytes. The adherence of a mannose-sensitive strain of E. coli to immobilized oral epithelial cells was inhibited by mannose derivatives. The adherence of fimbriated E. coli 987P to immobilized enterocytes was approximately four times higher than the adherence of a nonfimbriated variant of the same strain. The adherence of Streptococcus pyogenes to oral cells was detected in the range of 10 to 150 bacteria per cell and was inhibited by lipoteichoic acid and albumin. The data suggest that the putative receptors which bind bacteria on the immobilized cells retain a functional form similar to that of native cells in suspension. The proposed adherence assay is easy to perform, allows the detection of specific adherence of test bacteria, and provides objective quantitation of adherence with a sensitivity of 10 bacteria per cell. Most importantly, the assay allows the testing of many variables in the same day.

Animals↗