The difference in the IcrV sequences between Y. pestis and Y. pseudotuberculosis and its application for characterization of Y. pseudotuberculosis strains

Motin, Vladimir L.; Pokrovskaya, M. S.; Telepnev, Maksim V.; Кутырев, Владимир; Vidyaeva, Nadeshda A.; Filippov, Andrew A.; Smirnov, G. B.

doi:10.1016/0882-4010(92)90050-x

Cited by 30 publications

(31 citation statements)

References 23 publications

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“…This 37.3-kDa (327-amino acid residue) pure protein (Motin et al, 1992) is unstable and undergoes marked degradation during purification by traditional chromatographic procedures Motin et al, 1994). Nevertheless, LcrV expressed in E. coli in the form of fusion proteins engineered to contain the ligands staphylococcal Protein A (Motin et al, 1994;Nakajima et al, 1995), glutathione S-transferase (Leary et al, 1995), or sequences encoding polyhistidine Motin et al, 1996;Schmidt et al, 1999) can readily be purified to homogeneity by ligand affinity chromatography.…”

Section: Pcd-encoded Functionsmentioning

confidence: 96%

Yersinia pestis and Bubonic Plague

Brubaker¹

2006

The Prokaryotes

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Section: Pcd-encoded Functionsmentioning

confidence: 96%

Yersinia pestis and Bubonic Plague

Brubaker¹

2006

The Prokaryotes

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“…Passive transfer of both polyclonal and monoclonal anti-V Abs was shown previously to protect against lethal Y. pestis infection (2,20,30,34,40,46). In studies with polyclonal anti-V antisera adsorbed with different fragments of V, Motin et al showed that sera specific for the amino acids 168 to 275 (of the 326-amino-acid V) were protective (32). Hill et al cloned truncated fragments of V to help identify its antigenic and protective regions (20).…”

mentioning

confidence: 99%

Development of In Vitro Correlate Assays of Immunity to Infection withYersinia pestis

Bashaw

Norris

Weeks

et al. 2007

Clin Vaccine Immunol

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Pneumonic plague is a severe, rapidly progressing disease for which there is no effective vaccine. Since the efficacy of new vaccines cannot be tested in humans, it is essential to develop in vitro surrogate assays that are valid predictors of immunity. The F1 capsule antigen stimulates a protective immune response to most strains of Yersinia pestis. However, strains of Y. pestis that are F1؊ but still virulent have been isolated, and an in vitro assay, the results which can predict protection against both F1؉ and F1 ؊ strains, is needed. The virulence antigen (V) is an essential virulence factor of Y. pestis and stimulates protective antibodies. We investigated potential correlates of plague immunity that are based on anti-V antibodymediated neutralization of Yersinia-induced macrophage cytotoxicity. The neutralizing activity of sera from mice vaccinated with an F1-V fusion candidate vaccine was determined. The decrease in the level of the apoptosis-specific enzyme caspase-3 significantly predicted survival in one-and two-dose vaccination experiments. Sera from F1-V-vaccinated nonhuman primates were evaluated with macrophage assays based on caspase-3 and on other markers manifested at the different stages in cell death. Using murineand human-derived macrophages in microscopic and fluorescence-activated-cell-sorting-based live/dead staining assays of terminal necrosis, we demonstrated a strong association between in vitro neutralization of macrophage cytotoxicity induced by serum-treated Yersinia and in vivo protection against lethal infection. These results provide a strong base for the development of reliable in vitro correlate bioassays that are predictive of protective immunity to plague.Pneumonic plague is a severe and rapidly progressing disease for which no fully effective vaccine exists. There is no licensed vaccine available that elicits complete immunity in animal models to pneumonic plague. Although the previously licensed vaccine for plague protected experimental animals against parenteral challenge, it was ineffective against pneumonic challenge (19, 27; M. L. Pitt, unpublished data). Furthermore, the former vaccine, which consisted of killed whole cells of virulent Yersinia pestis (27), did not protect against virulent nonencapsulated (F1 Ϫ ) strains (19) since the vaccine did not contain immunogenic quantities of the virulence antigen (V) or other potentially protective immunogens (8,9,21). It was previously demonstrated that a combination of both F1 capsular protein antigen and V effectively protects against both encapsulated (F1 ϩ ) and nonencapsulated (F1 Ϫ ) strains of Y. pestis (3). New candidate plague vaccines containing a single recombinant F1-V fusion protein or a combination of these two proteins have been developed (19, 49a).The protective efficacy against lethal challenge of these new candidate plague vaccines cannot be ethically tested in humans. Thus, it is essential that an in vitro surrogate marker that can reliably predict the level of protective immunity in sera from vaccinated i...

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“…The probes were constructed by PCR amplification with primers directed at published sequences of the plasmidial genes caf1, encoding the structural sub-unit of the F1 antigen (10), pla encoding coagulase and the plasminogen activator (19) lcrV encoding the V antigen (16), and the chromosomal irp2 gene (11). The primer sequences used are described in Table 2.…”

Section: Methodsmentioning

confidence: 99%

Homology among extra-cryptic DNA bands and the typical plasmids in Brazilian Yersinia pestis strains

Leal

Sobreira

Leal

et al. 2000

Braz. J. Microbiol.

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Yersinia pestis, the etiologic agent of plague, harbors three well-characterized plasmids: pFra (90-110kb), pYV (70 kb) and pPst (9.5 kb). Furthermore, some extra-cryptic DNA bands have been observed in a number of wild strains from several foci of the world. Additional bands have also been reported in Brazilian strains. Looking for any relationship among these cryptic DNA bands and the three-prototypical plasmids, we analyzed twelve strains displaying different plasmid content. The DNA bands were hybridized by southern blot with probes directed at the genes caf1, lcrV and pla located respectively on the plasmids pFra, pYV and pPst. The probes were constructed by PCR amplification and labeled with digoxigenin. The Pla probe hybridized with its target (pPst) and with bands of about 35 kb suggesting some homology among them. The Caf1 probe hybridized with the target (pFra) as well as with higher bands. The LcrV also hybridized with the target (pYV) and both with the bands higher than pFra and the bands between pFra and pYV. These results suggest that the large-cryptic bands could represent some rearrangement, open circular or linearized forms of the pFra and pYV plasmids.

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The difference in the IcrV sequences between Y. pestis and Y. pseudotuberculosis and its application for characterization of Y. pseudotuberculosis strains

Cited by 30 publications

References 23 publications

Yersinia pestis and Bubonic Plague

Yersinia pestis and Bubonic Plague

Development of In Vitro Correlate Assays of Immunity to Infection withYersinia pestis

Homology among extra-cryptic DNA bands and the typical plasmids in Brazilian Yersinia pestis strains

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