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Biomedical subjects

S B Formal

Publications and source records attributed to S B Formal.

At least 37 records · Page 2Linked to original sources
PubMed1989

Electrolyte transport in rabbit cecum. I. Effect of RDEC-1 infection.

To investigate the characteristics of intestinal ion and fluid secretion induced by the adherent, effacing enteropathogenic Escherichia coli strain RDEC-1, we infected weanling rabbits with 10(7)-10(8) RDEC-1 organisms and then studied cecal ion transport under short-circuit conditions in Ussing chambers. Results in tissues with confluent adherent organisms were compared with those in uninfected ceca and in ceca stimulated with dibutyryl adenosine 3',5'-cyclic monophosphate (DBcAMP). The short-circuited cecum normally absorbed Na and Cl, secreted bicarbonate (as represented by the residual ion flux), and displayed a high rate of nondiffusional Na and Cl transport. RDEC-1 infection did not alter the short-circuit current (Isc), but it increased the conductance (Gt), decreased the potential difference (PD), abolished net Na absorption, and reversed Cl absorption to secretion. The changes in Na and Cl net fluxes may be explained by inhibition of a Na-Cl linked absorptive process. In contrast, DBcAMP significantly increased the Isc, PD, and Gt, decreased net Na flux, and abolished net Cl absorption by stimulating electrogenic Cl secretion. These results suggest that RDEC-1-induced changes in cecal ion transport are not mediated by cAMP. The reduction in Na-Cl linked absorption is consistent with anatomic changes in the apical surfaces of absorptive epithelial characteristic of effacing enteroadherence, whereas the increased conductance is consistent with tight junction disruption seen with RDEC-1 infection.

Biological Transport↗
PubMed1988

Deletion of the Shiga toxin gene in a chlorate-resistant derivative of Shigella dysenteriae type 1 that retains virulence.

We used a probe specific for detecting the structural-gene sequences of Shiga toxin to analyze the genetic nature of toxin synthesis in mutant derivatives of Shigella dysenteriae type 1. A chlorate-resistant (chl) mutant (725-78) of S. dysenteriae type 1 strain 3818T, which had retained virulence but had lost production of high levels of cytotoxic activity associated with Shiga toxin synthesis, contained a complete deletion of the Shiga toxin structural-gene sequences. These structural-gene sequences were also absent in a derivative of S. dysenteriae type 1 that contained a substitution of Escherichia coli DNA in the trp region of the chromosome. Isolates of Shigella flexneri and Shigella sonnei also did not react with the probe. The low-level cytotoxic activities associated with the mutant S. dysenteriae type 1 strains or with the virulent S. flexneri and S. sonnei strains are neutralizable with antiserum to Shiga toxin; however, these cytotoxic activities are not determined by the genes encoding classic Shiga toxin.

Bacterial Toxins↗
PubMed1987

Prevention of shigellosis by a Salmonella typhi-Shigella sonnei bivalent vaccine.

We genetically modified attenuated Salmonella typhi strain Ty21a to express the form I O polysaccharide antigen of Shigella sonnei. Three doses of this bivalent, live oral vaccine strain (1-8 X 10(9) organisms/dose) were given to young adults who, along with unvaccinated controls, were challenged one month later with pathogenic S. sonnei. The vaccinees had 40% protection against diarrhea and 56% against Hematest-positive diarrhea. Two of three vaccine lots provided higher levels of protection (53% against diarrhea and 71% against Hematest-positive diarrhea), but the third lot, prepared for a large-scale field trial, demonstrated no protective efficacy. Vaccinees had serum and local intestinal immune responses to S. sonnei lipopolysaccharide, and the presence of specific serum IgA or IgG antibody before challenge with pathogenic S. sonnei was correlated with protection from illness. Some lots of this bivalent vaccine strain provide significant protection against S. sonnei disease, but the problem of lot-to-lot variability must be overcome.

Adult↗
PubMed1987

Introduction of Shigella flexneri 2a type and group antigen genes into oral typhoid vaccine strain Salmonella typhi Ty21a.

For protection against dysentery caused by Shigella flexneri 2a, an in vivo-constructed recombinant plasmid with genes specifying the S. flexneri type and group antigens located near the pro (min 6) and his (min 44) chromosomal markers, respectively, was made and transferred to the galE Salmonella typhi strain Ty21a. Strain Ty21a carrying this recombinant plasmid was shown by immunological and biochemical analyses to express the S. flexneri 2a type and group antigens. Mice immunized with this vaccine strain were found to be protected against challenge with virulent S. flexneri 2a, but not significantly against S. typhi challenge, presumably because synthesis of the Shigella antigens interfered with expression of the typhoid antigens. Elimination of the recombinant plasmid from Ty21a allowed this strain to again express typical S. typhi O antigens. Mouse protection against both S. typhi and S. flexneri 2a challenges was achieved with a whole-cell vaccine mixture composed of equal parts of Ty21a and the Ty21a-S. flexneri 2a hybrid strain.

Administration, Oral↗
PubMed1986

Colonization, virulence, and mucosal interaction of an enteropathogenic Escherichia coli (strain RDEC-1) expressing shigella somatic antigen in the rabbit intestine.

The group factor 3,4 somatic antigen was transferred by recombination from a donor strain of Shigella flexneri type 2a to the Escherichia coli O15 rabbit pathogen strain RDEC-1. A hybrid clone (7482-1-1) that expressed only the S. flexneri 3,4 somatic antigen and a second clone (7482-1-7) from the same mating mixture that expressed only the O15 somatic antigen were compared for virulence in rabbits. The 7482-1-1 strain produced diarrhea in 10% of rabbits versus 84% for the 7482-1-7 strain (P less than .001). In fluorescent antibody-stained, frozen, sectioned tissues, fewer 7482-1-1 bacteria adhered to ileum (P = .013) and cecum (P = .044). Light and electron microscopic studies demonstrated that the recombinant strain, which adhered to membranous (M) cells to the same degree as the 7482-1-7 strain, was found beneath the epithelium, where its presence resulted in a marked acute inflammatory response. Both the 7482-1-1 and 7482-1-7 strains exhibited characteristic close adherence to M cells and absorptive ileal and cecal mucosa.

Animals↗
PubMed1986

Effect of guinea pig or monkey colonic mucus on Shigella aggregation and invasion of HeLa cells by Shigella flexneri 1b and 2a.

The effects of guinea pig and rhesus monkey colonic mucus preparations on Shigella aggregation and invasion of HeLa cell monolayers by Shigella flexneri serotype 1b, 2a, and 5 strains were investigated. Guinea pig mucus caused agglutination of S. flexneri serotype 1b but not of S. flexneri serotype 2a or 5. Guinea pig mucus also inhibited HeLa cell invasion by S. flexneri serotypes 1b and 2a. Monkey mucus neither agglutinated any Shigella strain nor inhibited HeLa cell invasion.

Adhesiveness↗
PubMed1986

Serum immune response to Shigella protein antigens in rhesus monkeys and humans infected with Shigella spp.

The serum antibody response to proteins encoded by the virulence-associated plasmid of Shigella flexneri was determined in monkeys challenged with virulent S. flexneri serotype 2a. With water-extractable antigen in an enzyme-linked immunosorbent assay, a significant increase in antibody titer against proteins from a plasmid-carrying, virulent strain of S. flexneri serotype 5 could be demonstrated in convalescent sera. There were minimal antibody titers against proteins from an avirulent (plasmid-free) organism. Previously identified plasmid-coded polypeptides a, b, c, and d were predominant antigens recognized by a majority of the convalescent sera in immunoblots. An additional 140-megadalton plasmid-coded polypeptide was also recognized by half of the sera. Convalescent serum from an infected monkey recognized antigens on the bacterial surface in several different plasmid-containing Shigella species and in an enteroinvasive Escherichia coli strain. A survey of sera obtained from children 5 to 10 years of age who had been infected with S. flexneri or S. sonnei revealed high enzyme-linked immunosorbent assay titers in both acute and convalescent sera against a water extract from a virulent Shigella strain. In contrast, children under 3 years of age had no antibody titer in either acute or convalescent sera against the virulence-associated shigella proteins, while 3- to 4-year-old children mounted an immune response against these proteins only in convalescence.

Animals↗
PubMed1986

Secretory immunoglobulin A response following peroral priming and challenge with Shigella flexneri lacking the 140-megadalton virulence plasmid.

This study evaluates the ability of noninvasive Shigella spp., lacking the 140-megadalton virulence plasmid, to elicit a mucosal immunoglobulin A immune response in the intestine. For these studies, we used Shigella flexneri M4243A1 (which lacks the plasmid and is Sereny test negative) to prime and challenge three groups of rabbits perorally. Both primary and immunoglobulin A memory responses were detectable in these secretions. These findings indicate that a mucosal memory response can be primed by nonpathogenic strains of Shigella which lack the virulence plasmid.

Administration, Oral↗
PubMed1986

Morphologic evaluation of the effects of Shiga toxin and E coli Shiga-like toxin on the rabbit intestine.

The effects of a Shiga toxin derived from Shigella dysenteriae Type 1, Strain 60R, and a Shiga-like toxin from the enterohemorrhagic Escherichia coli O157:H7, Strain 933, were studied in the in vivo rabbit ileal loop model. The effects of both toxins were similar and resulted in severe villus blunting by 18-24 hours after exposure. With both toxins, a dose effect was noted; and the lesions, first detected at 2 hours after inoculation, became more severe over time. Both toxins appeared to act directly and selectively on the mature columnar absorptive epithelium of the intestinal villus, which resulted in the premature expulsion of these cells from the lateral villus wall, with a decrease in the villus/crypt ratio. The goblet mucous cells remained attached and frequently formed clusters on the blunt villus apices. The crypt epithelium underwent a rapid proliferation and maintained the epithelial integrity. The ultrastructural changes observed in the toxin-injured villus absorptive cells suggested that these cells underwent a process of apoptosis, rather than necrosis. These findings suggest that both toxins act in vivo in the small intestine on a specific cell population, the mature, differentiated absorptive villus epithelium.

Animals↗
PubMed1985

Plaque formation by virulent Shigella flexneri.

An in vitro tissue culture plaque assay was developed to investigate the intracellular replication and intercellular spread of virulent shigellae. Shigella plaques were formed in HeLa cell monolayers in the presence of an agarose overlay containing tissue culture medium and gentamicin, which eliminated extracellular bacterial growth. Microscopically, the plaques were characterized by a central area of dead host cells surrounded by cells infected with shigellae. Cells further away from the plaque center were uninfected. Inclusion of chloramphenicol or nalidixic acid in the overlay completely abolished plaque formation. Plaque formation was completely inhibited when infected monolayers were shifted from 37 to 30 degrees C. Shifting infected monolayers from 30 degrees C, where plaques do not form, to 37 degrees C resulted in the formation of plaques. Cultures of Shigella boydii, Shigella sonnei (form I), and all six serotypes of Shigella flexneri produced plaques. Shigellae isolated from plaques were Sereny test positive, contained a 140-megadalton plasmid, and were gentamicin sensitive. Noninvasive shigellae did not form plaques.

Adsorption↗
PubMed1985

Synthesis of aerobactin and a 76,000-dalton iron-regulated outer membrane protein by Escherichia coli K-12-Shigella flexneri hybrids and by enteroinvasive strains of Escherichia coli.

One of the chromosomal segments associated with the virulence of Shigella flexneri and transferred to Escherichia coli K-12 by conjugation has been shown to code for the production of aerobactin and for the synthesis of an iron-regulated 76,000-dalton (76K) outer membrane protein. Analysis of various E. coli K-12-S. flexneri transconjugants showed that the genes involved with the synthesis of aerobactin and with the production of the 76K protein were linked to the mtl region of the S. flexneri chromosome. S. flexneri itself synthesized a 76K protein in its outer membrane under iron restriction as well as traces of 81K and 74K proteins. An examination of four enteroinvasive strains of E. coli showed that each produced aerobactin and a 76K outer membrane protein during iron-restricted growth. The profile of the iron-regulated proteins expressed by the enteroinvasive strains of E. coli was virtually identical to that expressed by the laboratory-constructed E. coli K-12-S. flexneri hybrids under the same growth conditions.

Animals↗
PubMed1985

Identification and antigenic characterization of virulence-associated, plasmid-coded proteins of Shigella spp. and enteroinvasive Escherichia coli.

Seven plasmid-coded polypeptides, designated a through g, were identified by two-dimensional nonequilibrium pH gradient electrophoresis of radiolabeled extracts from minicells of virulent Shigella flexneri serotypes 2a and 5 and enteroinvasive Escherichia coli O143. These polypeptides were deemed to be products of 140-megadalton (MDa) virulence-associated plasmids because they were not synthesized in minicells which were not harboring a 140-MDa plasmid or in minicells which were carrying an F lac plasmid of the same incompatibility group. Synthesis of these polypeptides was repressed in minicells incubated at 30 degrees C and in minicells isolated from a noninvasive opaque colonial variant, even though these strains harbored a 140-MDa plasmid. Enriched fractions of polypeptides b, c, and d were obtained from S. flexneri serotype 5 by preparative isoelectric focusing, and polyclonal rabbit antisera recognizing each polypeptide were raised. These antisera were able to detect cross-reacting plasmid-coded polypeptide antigens in S. flexneri serotype 3, Shigella sonnei, and enteroinvasive E. coli O143. In addition, Western blots of minicell extracts from S. flexneri serotype 5 or E. coli O143 indicated that plasmid-coded polypeptides a through d were recognized by convalescent antiserum from a monkey infected with S. flexneri serotype 2a.

Animals↗
PubMed1984

Shiga-like toxin-converting phages from Escherichia coli strains that cause hemorrhagic colitis or infantile diarrhea.

Escherichia coli K-12 acquired the ability to produce a high titer of Shiga-like toxin after lysogenization by either of two different bacteriophages isolated from a highly toxinogenic Escherichia coli O157:H7 strain that causes hemorrhagic colitis. One of these phages and another Shiga-like toxin-converting phage from an Escherichia coli O26 isolate associated with infantile diarrhea were closely related in terms of morphology, virion polypeptides, DNA restriction fragments, lysogenic immunity, and heat stability, although a difference in host range was noted. These phages are currently the best-characterized representatives from a broader family of Shiga-like toxin-converting phages.

Animals↗