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Biomedical subjects

S B Formal

Publications and source records attributed to S B Formal.

At least 55 records · Page 3Linked to original sources
PubMed1984

Unusual lipopolysaccharide antigens of a Salmonella typhi oral vaccine strain expressing the Shigella sonnei form I antigen.

Salmonella typhi 5076-1C, a potential live, oral vaccine for protection against typhoid fever and Shigella sonnei shigellosis, expresses the S. sonnei form I antigen and normal S. typhi somatic antigens. Polysaccharide antigens of this galactose epimeraseless genetic derivative strain were hot phenol-water extracted from cells grown with (+gal) and without (-gal) galactose. Ultracentrifugation of the aqueous layer from (+gal) cells resulted in a lipopolysaccharide (LPS) pellet having core-linked S. typhi O-antigen but no core-linked form I antigen; the LPS from (-gal) cells lacked O-antigen. The form I antigen, obtained from the supernatant, was purified by alcohol precipitation and ion exchange chromatography. Unlinked form I and S. typhi O-polysaccharide antigens, both present in the (+gal) supernatant, were further separated by gel filtration. Chemical analyses revealed the 5076-1C form I antigen to be a polymer (Mr = 14,000-20,000) having O-disaccharide repeating units comprised of 2-acetamido-4-amino-2, 4,6-trideoxy-D-galactose and 2-acetamido-2-deoxy-L-altruronic acid. Unlike parental S. sonnei form I LPS, the 5076-1C form I antigen lacked core lipid A, had low phosphorus content, and migrated in polyacrylamide gels with lower relative mobility. In contrast to current concepts of LPS assembly, these data indicate that 5076-1C form I antigen is transported to the cell surface without covalent linkage to core lipid A, and exists as a polymerized, antigenic surface entity.

Administration, Oral↗
PubMed1984

Safety and antigenicity of typhoid-Shigella sonnei vaccine (strain 5076-1C).

The form-1 antigen of Shigella sonnei was transferred to the avirulent Salmonella typhi strain 21a and the resulting 5076-1C transconjugate strain was tested for safety and immunogenicity as a candidate oral vaccine. The transconjugant strain was shown to be well tolerated and safe in 19 human volunteers who were fed from 1 X 10(7) to 1 X 10(10) organisms. Only two of 10 volunteers tested had developed a rise in antibody titer to the lipopolysaccharide of the hybrid 5076-1C strain.

Administration, Oral↗
PubMed1984

Oral vaccination of monkeys with an invasive Escherichia coli K-12 hybrid expressing Shigella flexneri 2a somatic antigen.

A living oral vaccine, designed to protect against Shigella flexneri 2a infections, was constructed by using Escherichia coli K-12 as a carrier strain. The hybrid strain, designated EC104, contained both chromosomal and plasmid genes from S. flexneri donor strains. In addition to expressing the S. flexneri 2a somatic antigen, it had inherited the property of epithelial-cell invasion. After the oral administration to rhesus monkeys, EC104 was isolated from the feces for up to 3 days, but by day 4 all stool cultures were negative. The serum antibody response against S. flexneri 2a somatic antigen was variable, but the vaccine conferred significant protection against an oral challenge with virulent S. flexneri 2a.

Administration, Oral↗
PubMed1984

Expression of lipopolysaccharide O antigen in Escherichia coli K-12 hybrids containing plasmid and chromosomal genes from Shigella dysenteriae 1.

The requirement for both plasmid and chromosomal genes in the biosynthesis of Shigella dysenteriae 1 lipopolysaccharide O antigen was demonstrated in Escherichia coli-Shigella hybrids. A 6-megadalton S. dysenteriae 1 plasmid, designated pWR23, was phenotypically tagged with the Tn3 ampicillin-resistance transposon. The tagged plasmid, designated pWR24, was transferred by transformation or conjugal mobilization to a rough E. coli K-12 recipient. Although the resultant hybrids were agglutinated in S. dysenteriae 1 antiserum, they did not remove all of the anti-Shiga agglutinins in absorption experiments. Modified lipid A core structure was detected in these hybrids, but Shiga O antigen was not expressed. When the his+ locus of the S. dysenteriae 1 chromosome was transferred by transduction to E. coli K-12 containing pWR24, complete Shiga O antigen was expressed. Lipopolysaccharide extracted from these hybrids was indistinguishable chemically, electrophoretically, and serologically from native S. dysenteriae 1 lipopolysaccharide.

Antigens, Bacterial↗
PubMed1983

Genetic transfer of a mucosal adherence factor (R1) from an enteropathogenic Escherichia coli strain into a Shigella flexneri strain and the phenotypic suppression of this adherence factor.

Escherichia coli strain RDEC-1 avidly adheres to rabbit ileal brush borders. Two separate experiments were designed to determine whether pili promote this adherence. (1) Adherence of strain RDEC-1 was phenotypically suppressed by changing the culture medium. Loss of adherence was correlated with the absence of pili. Thus, growth of strain RDEC-1 in Penassay broth (Difco Laboratories, Detroit, Mich.) promoted both adherence and expression of pili on greater than or equal to 90% of organisms, whereas growth in brain-heart infusion medium suppressed adherence and reduced the percentage of piliated organisms to less than or equal to 13%. (2) The adherence ability of strain RDEC-1 was genetically transferred to previously nonadherent and nonpiliated Shigella flexneri. The Shigella exconjugants that inherited the adherence ability were uniformly piliated, while all nonadherent Shigella exconjugants were nonpiliated. Finally, the pili on both RDEC-1 and the Shigella exconjugant strains were shown to be distinct from type 1 pili. Therefore, unique pili confer upon strain RDEC-1 the ability to adhere to rabbit intestinal brush borders.

Adhesiveness↗
PubMed1983

Alterations in the pathogenicity of Escherichia coli K-12 after transfer of plasmid and chromosomal genes from Shigella flexneri.

A 140-megadalton plasmid (pWR110), which has previously been associated with virulence in Shigella flexneri, was transferred to Escherichia coli K-12. Segments of S. flexneri chromosomal material were then transferred to the plamid-bearing K-12 strains. The virulence of these transconjugant hybrids was assessed in the HeLa cell model, in ligated rabbit ileal loops, or in the Sereny test. A K-12 strain which harbored only pWR110 invaded HeLa cells, produced minimal lesions in the rabbit ileal mucosa, and was negative in the Sereny test. Plasmid-containing K-12 hybrids which had incorporated various shigella chromosomal regions gave differential reactions in the rabbit ileal loops and in the Sereny test. Analysis of these transconjugants indicated that three regions were linked with virulent phenotypes. These included the his region (when the genes responsible for O-antigen synthesis were cotransferred) and the kcp locus (linked to the lac-gal region). Either of these chromosomal regions was sufficient to allow invasion of the rabbit ileal mucosa. In addition to both of these regions, another shigella chromosomal segment linked to the arg and mtl loci was necessary for fluid production in the rabbit ileal loop and for a positive Sereny reaction. Thus, derivatives of an E. coli K-12 strain, constructed by the stepwise conjugal transfer of a large plasmid and three chromosomal segments from S. flexneri, appeared to contain the necessary determinants for full pathogenicity in a variety of laboratory models.

Animals↗
PubMed1983

Characterization of virulence plasmids and plasmid-associated outer membrane proteins in Shigella flexneri, Shigella sonnei, and Escherichia coli.

The 140-megadalton plasmids of Shigella flexneri serotypes 1, 3, and 5, in addition to the 120-megadalton plasmid of Shigella sonnei, are associated with virulence. The present study showed that a 140-megadalton plasmid is also associated with virulence in Escherichia coli. When these plasmids were cleaved with EcoRI or BamHI restriction endonucleases, considerable homology was evident in plasmids from S. sonnei strains, whereas only a few common fragments were observed among the S. flexneri and enteroinvasive E. coli plasmids. Nitrocellulose filter hybridization demonstrated that, despite variations in restriction sites, all these plasmids shared a considerable complement of homologous sequences. Minicell-producing strains were obtained by N-methyl-N'-nitro-N-nitrosoguanidine mutagenesis. Transmission electron microscopy of infected HeLa cells showed that minicells from invasive strains retained the invasive phenotype. Sixteen polypeptides were labeled when S. flexneri 5 minicells were incubated with [35S]methionine. Fourteen of these plasmid-coded polypeptides were associated with the outer membrane in invasive strains of S. flexneri 5, and nine polypeptides of similar molecular weight were labeled in the outer membrane of invasive strains of S. flexneri 3, S. sonnei, and E. coli. Seven of the S. flexneri 5 polypeptides were not labeled in a noninvasive strain which had sustained a large deletion in the virulence-associated plasmid, and none were labeled in minicells which no longer harbored this plasmid.

Animals↗
PubMed1982

Production of Shigella dysenteriae type 1-like cytotoxin by Escherichia coli.

Strains of Escherichia coli previously implicated or proven to be causes of diarrhea were examined for production of a toxin similar to that of Shigella dysenteriae type 1 (Shiga). Organisms grown in an iron-depleted broth were lysed by pressure disruption followed by ultracentrifugation. Saline-dialyzed extracts were tested for cytotoxic effects on HeLa cells that were neutralizable with antiserum to Shiga toxin. Among the 13 E. coli strains so analyzed, 11 made a Shiga-like cytotoxin in levels ranging from trace (two avirulent isolates) to amounts equivalent to S. dysenteriae type 1 (two noninvasive strains that did not make E. coli heat-labile or -stable enterotoxins but were isolated from infants with diarrhea). As with extracts of Shiga toxin, lysates of these E. coli strains that produced high levels of Shiga-like toxin were enterotoxic for rabbits, paralytic and lethal for mice, and inhibited protein synthesis in HeLa cells. Thus, these data suggest that Shiga-like toxin may be another heretofore undiscovered factor in the pathogenesis of diarrhea caused by some E. coli strains.

Animals↗
PubMed1982

Involvement of a plasmid in the invasive ability of Shigella flexneri.

Representative Shigella flexneri strains were studied to determine whether plasmids are involved in their virulence. All invasive S. flexneri strains, irrespective of serotype, were found to harbor a large plasmid of approximately 140 megadaltons in size, although some strains carried additional plasmid species. Spontaneous variants of strains of serotypes 1, 2, and 5 had lost this 140-megadalton plasmid and had concomitantly become avirulent, i.e., could neither invade HeLa cell monolayers nor produced keratoconjunctivitis in guinea pigs. To monitor plasmid transfer, the 140-megadalton plasmid of strain M90T (serotype 5) was tagged with the kanamycin resistance transposon Tn5. This tagged plasmid, pWR110, was not self-transmissible, but was mobilized by one of several different conjugative plasmids into avirulent derivatives of the heterologous serotypes 1 and 2 which had lost the comparable large plasmid. Transconjugants of both serotypes which had received pWR110 regained virulence. These data directly demonstrate that this large S. flexneri plasmid encodes or regulates some function(s) required for epithelial cell penetration.

Animals↗
PubMed1982

Intestinal immunoglobulin A responses in rabbits to a Salmonella typhi strain harboring a Shigella sonnei plasmid.

Salmonella typhi 5076-IC, which contains a plasmid that encodes the form I antigen of Shigella sonnei and which expresses S. typhi 9 and 12 and S. sonnei form I antigens, was used to immunize rabbits via chronically isolated ileal loops. Intestinal immunoglobulin A activity was detected against S. typhi, S. sonnei form I, and S. typhi strain 5076-IC. Thus S. typhi 5076-IC can effectively elicit mucosal immunoglobulin A to both S. typhi and S. sonnei.

Animals↗
PubMed1981

Role of antigen form in development of mucosal immunoglobin A response to Shigella flexneri Antigens.

One major stumbling block in the development of an effective means to immunize against shigellosis and other enteric diseases has been the lack of a means to assess sequential mucosal immune responses to different potential immunogens. In the present study, we compared the abilities of live invasive organisms, noninvasive organisms, and nonviable antigen preparations of shigella to elicit mucosal immune responses. Whereas previous studies have found that effective immunity was produced best by vaccination with live invasive strains of shigella, in the present study, live noninvasive strains that did not produce any histopathological damage were consistently able to produce local (immunoglobulin A) immune responses as vigorous as those of the invasive strains. Further, acetone-killed shigella antigen was also an effective mucosal immunogen, whereas hot phenol-water-extracted shigella lipopolysaccharide was ineffective, possibly due to the method of preparation. A single oral or parenteral priming was ineffective in enhancing the mucosal immune response when restimulated 1 month later with the same antigen. However, a mucosal memory response was found to be present several months after a triple mucosal stimulation with a locally invasive vaccine strain.

Administration, Oral↗
PubMed1981

Protein synthesis in HeLa or Henle 407 cells infected with Shigella dysenteriae 1, Shigella flexneri 2a, or Salmonella typhimurium W118.

The incorporation of [14C]leucine into protein was studied in two mammalian cell lines which had been infected with strains of Shigella dysenteriae 1, Shigella flexneri 2a, or Salmonella typhimurium W118. These cell lines differed in susceptibility to the effects of exogenously applied Shiga cytotoxin. All invasive shigella strains (which synthesize this toxin to a greater or lesser degree) were found to inhibit protein synthesis in both cell lines with equal efficiency. Leucine accumulation continued in these cells, but the labeled amino acid was preferentially incorporated into bacterial protein. S. typhimurium W118, which has not been shown to elaborate a Shiga-like toxin, had little effect on protein synthesis in infected host cells.

Bacterial Proteins↗
PubMed1981

Shigella sonnei plasmids: evidence that a large plasmid is necessary for virulence.

Virulent form I Shigella sonnei strains contain a 120-megadalton plasmid that is absent in their form II derivatives, which are always avirulent and devoid of O side chains. In the present study, 165 biochemical and antibiotic traits were assessed, but no experimentally useful phenotype could be associated with this large form I plasmid. Therefore, the form I plasmids of several S. sonnei strains were tagged with the antibiotic resistance transposons Tn3, Tn5, or Tn10. Transposon-tagged form I plasmids were not self-transmissible, but could be mobilized by the plasmid R386. Form II S. sonnei transconjugants for the form I plasmid acquired both virulence and the ability to synthesize form I antigen, establishing that these properties are plasmid mediated. Further studies indicate that this 120-megadalton form I plasmid is physically unstable in any of several host bacteria and suggest that it is a member of the FI incompatibility group. Also, two commonly observed, small plasmids of S. sonnei, of 3.2 and 3.9 megadaltons, were shown to encode either colicin E1 production or resistance to streptomycin and sulfonamide, respectively.

Anti-Bacterial Agents↗
PubMed1981

Construction of a potential bivalent vaccine strain: introduction of Shigella sonnei form I antigen genes into the galE Salmonella typhi Ty21a typhoid vaccine strain.

Shigella sonnei, an intestinal pathogen, produces a characteristic form I cell surface antigen now known to be plasmid encoded. We considered that the GalE Salmonella typhi Ty21a oral vaccine strain, highly effective against typhoid, might be modified so as to be protective also against shigellosis due to S. sonnei. The plasmid responsible for form I antigen synthesis was therefore conjugally transferred to the galE S. typhi strain. Serological studies revealed that the derivative strain produces the form I antigen in addition to the normal S. typhi somatic antigens. Testing in mice demonstrated that the derivative form I galE S. typhi strain is protective against both S. sonnei and S. typhi challenges. These data suggest that the galE S. Ty21a oral vaccine strain, which presumably stimulates the local immune system in the intestine, may also serve as a useful carrier for other antigenic determinants to protect against different intestinal infections.

Animals↗
PubMed1980

Cytotoxicity of Shigella dysenteriae 1 for cultured mammalian cells.

The cytotoxicity of an invasive toxigenic wild-type strain of Shigella dysenteriae 1 (3818T) was compared with that of noninvasive, toxigenic strain 38180 and hypotoxigenic strain 725. Cytolysis of HeLa or Henle 407 cells exposed to these strains was measured by release of (3H) uridine from prelabeled monolayers. HeLa cells exposed to noninvasive, toxigenic strain 38180, or to partially purified Shiga toxin were lysed only after a latent period of more than 8 hr. During this period, protein synthesis was inhibited. In contrast, Henle 407 cells that were exposed to strain 38180 or to exogenous Shiga toxin were unaffected. When either Henle 407 or HeLa cells were infected with invasive toxigenic strains, rapid lysis ensued. Quantitative microassay of cytosol toxicity showed that Shiga toxin was produced intracellularly by strain 3818T. The data suggest that cytolysis of infected mammalian cells is caused, at least in part, by intracellular Shiga toxin.

Bacterial Toxins↗