Characterization of the Shigella serotype D (S. sonnei) O polysaccharide and the enterobacterial R1 lipopolysaccharide core by use of mouse monoclonal antibodies

Viret, Jean-François; Bruderer, Urs; Lang, Aloïs B.

doi:10.1128/iai.60.7.2741-2747.1992

Cited by 12 publications

(6 citation statements)

References 34 publications

(37 reference statements)

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“…VOL. 64,1996 CARRIER-BASED S. SONNEI LIVE ORAL VACCINES contain rfaD in addition to the complete rfb locus, the lethal effect may instead be related to a defect in LPS core biosynthesis. If such is the case, the lethality of this particular deletion in the CH9 background would indicate that the integrated rfa R1 locus cannot complement the defect.…”

Section: Discussionmentioning

confidence: 99%

“…Determination of anti-LPS antibodies. The sera from mice were individually tested for the presence of LPS-specific antibodies with purified S. sonnei phase 1 or V. cholerae Inaba LPS as immobilized antigens in an ELISA as previously described (64).…”

Section: Methodsmentioning

confidence: 99%

“…Since we could not produce a CH3 deletion mutant using pSSVI255-20, a genotypically similar strain, CH22, was constructed by the reverse approach, namely the integration of rfb/rfc sonnei genes carried on plasmid pSSVI201-1 into the VOL. 64,1996 CARRIER-BASED S. SONNEI LIVE ORAL VACCINES chromosome of CH19 (Fig. 3).…”

Section: Cloning Of the Rfb Inaba Locus From Cvd103-hgr A Sau3amentioning

confidence: 99%

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Development of Shigella sonnei live oral vaccines based on defined rfbInaba deletion mutants of Vibrio cholerae expressing the Shigella serotype D O polysaccharide

1996

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Previous experimentation has highlighted a number of difficulties in the development of carrier-based bivalent vaccines (J.-F. Viret and D. Favre, Biologicals 22:361-372, 1994). In an attempt to obviate these problems, we decided to express the Shigella sonnei serotype D O polysaccharide (O-PS) in rough Vibrio cholerae carrier strains. Toward this aim, a series of defined rfb Inaba deletion (⌬rfb Inaba) mutants of the cholera vaccine strain V. cholerae CVD103-HgR (O1 Inaba serotype) and derivatives bearing the chromosomally integrated locus encoding the S. sonnei O-PS were constructed and characterized. The various mutations disrupt genes thought to be involved in either the synthesis of perosamine, the synthesis of 3-deoxy-L-glycero tetronic acid, or the O-PS transport functions together with synthesis of the perosamine synthetase. Some deletions were obtained only in strains expressing the heterologous lipopolysaccharide (LPS). Viable ⌬rfb Inaba deletions in CVD103-HgR profoundly altered some of its phenotypic properties. The same deletions present in CVD103-HgR derivatives expressing the heterologous LPS affected their phenotypes only to a lesser extent. Only in strains in which perosamine synthesis was specifically abolished could high amounts of core-bound S. sonnei O-PS be synthesized. Two such strains (CH21, which expresses both the R1 core and the S. sonnei O-PS, and CH22, which expresses only the latter antigenic determinant) were further analyzed and were found to be indistinguishable from CVD103-HgR with regard to lack of enterotoxin activity, choleragenoid production, mercury resistance, pilin production, and, for CH22, motility. Mice immunized with CH22 produced high titers of S. sonnei O-PS-specific antibodies.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Cloning Of the Rfb Inaba Locus From Cvd103-hgr A Sau3amentioning

confidence: 99%

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Development of Shigella sonnei live oral vaccines based on defined rfbInaba deletion mutants of Vibrio cholerae expressing the Shigella serotype D O polysaccharide

1996

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“…SDS-15% PAGE gels, immunoblot analyses of LPS preparations, and ELISA were performed as described previously (Viret et al, 1992). For SDS-PAGE analysis, preparations were loaded in parallel with a pre-stained protein molecularweight standard (low range; Bio-Rad).…”

Section: Sds-page Immunoblots and Elisamentioning

confidence: 99%

“…First antibodies used for the immunoblot analysis of LPS preparations were the S. sonnei O-PS-specific mAb Sh5S and the R1 core-specific mAb Sh9R (Viret et al, 1992), as well as the V. cholerae Inaba/Ogawa O-PS-specific mAb VCO4 (previously termed H4; Gustafsson and Holme, 1983) (provided by D. Moss, Wellcome Research Lab., Beckenham, UK). Peroxidase-conjugated affinity-isolated goat anti-mouse IgM antibodies ( Tago) were used as developing antibodies.…”

Section: Sds-page Immunoblots and Elisamentioning

confidence: 99%

Expression of Shigella sonnei lipopolysaccharide in Vibrio cholerae

Viret

Cryz

Favre

1996

Molecular Microbiology

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Making use of a newly designed mobilizable suicide vector, the genetic determinants encoding Shigella sonnei lipopolysaccharide (LPS) were stably integrated into the chromosome of the live attenuated Vibrio cholerae vaccine strain CVD103-HgR. Expression studies showed that the production of complete S. sonnei O-polysaccharide (O-PS)-bearing LPS was limited in bivalent recombinant strains that were also proficient in the synthesis of the host-encoded Inaba O-PS. Conversely, high amounts of LPS carrying S. sonnei O-PS are produced in monovalent Inaba-deficient derivatives, even in those strains which do not co-express the compatible R1 LPS core. Thus, the non-enterobacterial V. cholerae LPS core efficiently acts as a receptor for covalent binding of S. sonnei O-PS provided that competition with the host O-PS is avoided. Expression of the R1 core interferes with cell division in recombinant V. cholerae without affecting other physiological properties of vaccine strain CVD103-HgR. Both monovalent and bivalent strains stimulated high serum-antibody titres specific for their respective O-serotype(s) when administered to rabbits. The potential of V. cholerae as an expression carrier for heterologous O-serotypes is discussed.

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Antibodies to lipopolysaccharide

Poxton

1995

Journal of Immunological Methods

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Characterization of the Shigella serotype D (S. sonnei) O polysaccharide and the enterobacterial R1 lipopolysaccharide core by use of mouse monoclonal antibodies

Abstract: In the course of developing a live vaccine, we generated three murine monoclonal antibodies (MAb) specific

Cited by 12 publications

References 34 publications

Development of Shigella sonnei live oral vaccines based on defined rfbInaba deletion mutants of Vibrio cholerae expressing the Shigella serotype D O polysaccharide

Development of Shigella sonnei live oral vaccines based on defined rfbInaba deletion mutants of Vibrio cholerae expressing the Shigella serotype D O polysaccharide

Expression of Shigella sonnei lipopolysaccharide in Vibrio cholerae

Antibodies to lipopolysaccharide

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