Laboratory Maintenance and Characterization of <i>Yersinia pestis</i>

Bearden, Scott W.; Perry, Robert D.

doi:10.1002/9780471729259.mc05b01s11

Cited by 8 publications

(3 citation statements)

References 19 publications

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“…pestis has been classified as a Category A select agent (8) requiring special precautions, particularly because of the risk of pneumonic plague due to bacterial aerosolization. One of the major impediments to studying the biology and pathogenesis of Y. pestis is that many laboratories do not have access to the required biosafety level 3 (BSL-3) facilities and therefore must work with attenuated strains that were excluded from the CDC select-agents list.…”

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confidence: 99%

Biosafety Level 2 Model of Pneumonic Plague and Protection Studies with F1 and Psa

et al. 2010

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Attenuated Yersinia pestis pgm strains, such as KIM5, lack the siderophore yersiniabactin. Strain KIM5 does not induce significant pneumonia when delivered intranasally. In this study, mice were found to develop pneumonia after intranasal challenge with strain KIM5 when they were injected intraperitoneally with iron dextran, though not with iron sulfate. KIM5-infected mice treated daily with 4 mg iron dextran died in 3 days with severe pneumonia. Pneumonia was less severe if 4 mg iron dextran was administered only once before infection. The best-studied experimental vaccine against plague currently consists of the Yersinia pestis capsular antigen F1 and the type 3 secreted protein LcrV. The F1 antigen was shown to be protective against KIM5 infections in mice administered iron dextran doses leading to light or severe pneumonia, supporting the use of an iron dextran-treated model of pneumonic plague. Since F1 has been reported to be incompletely protective in some primates, and bacterial isolates lacking F1 are still virulent, there has been considerable interest in identifying additional protective subunit immunogens. Here we showed that the highly conserved Psa fimbriae of Y. pestis (also called pH 6 antigen) are expressed in murine organs after infection through the respiratory tract. Studies with iron dextran-treated mice showed that vaccination with the Psa fimbrial protein together with an adjuvant afforded incomplete but significant protection in the mouse model described. Therefore, further investigations to fully characterize the protective properties of the Psa fimbriae are warranted.

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confidence: 99%

Biosafety Level 2 Model of Pneumonic Plague and Protection Studies with F1 and Psa

et al. 2010

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“…Since Y. pestis is classified as a category A select agent by the CDC [23] , manipulation of wt Y. pestis requires a Biosafety Level 3 (BSL-3) facility, which limits access to and work with this pathogen. Thus, in our study, we established a platform for analyzing Y. pestis virulence factors by means of the wt BSL-2 pathogen, S. Typhimurium.…”

Section: Discussionmentioning

confidence: 99%

Vulnerabilities in Yersinia pestis caf Operon Are Unveiled by a Salmonella Vector

et al. 2012

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During infection, Yersinia pestis uses its F1 capsule to enhance survival and cause virulence to mammalian host. Since F1 is produced in large quantities and secreted into the host tissues, it also serves as a major immune target. To hold this detrimental effect under proper control, Y. pestis expresses the caf operon (encoding the F1 capsule) in a temperature-dependent manner. However, additional properties of the caf operon limit its expression. By overexpressing the caf operon in wild-type Salmonella enterica serovar Typhimurium under a potent promoter, virulence of Salmonella was greatly attenuated both in vitro and in vivo. In contrast, expression of the caf operon under the regulation of its native promoter exhibited negligible impairment of Salmonellae virulence. In-depth investigation revealed all individual genes in the caf operon attenuated Salmonella when overexpressed. The deleterious effects of caf operon and the caf individual genes were further confirmed when they were overexpressed in Y. pestis KIM6+. This study suggests that by using a weak inducible promoter, the detrimental effects of the caf operon are minimally manifested in Y. pestis. Thus, through tight regulation of the caf operon, Y. pestis precisely balances its capsular anti-phagocytic properties with the detrimental effects of caf during interaction with mammalian host.

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“…An avirulent strain, A1122, of Y. pestis was procured from Dr. Scott Bearden, Bacterial Diseases Branch, Centers for Disease Control and Prevention (CDC; Fort Collins, CO) (22,23). Glycerol stocks of Y. pestis were streaked on tryptic soy agar (TSA) plates with colonies appearing after 48 h, when incubated at 30°C.…”

Section: Test Organism Preparationmentioning

confidence: 99%

Investigations into Enhancing Yersinia pestis Cells Viability following Environmental Sampling for Forensic Analysis

Rastogi

Smith

Burton

et al. 2020

Journal of Forensic Sciences

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Following an intentional or accidental bio‐warfare agent (BWA) release, environmental sample analysis is absolutely critical to determine the extent of contamination. When dealing with nonspore forming BWA (e.g., Yersinia pestis), retention of cell viability is central to such analyses. Even though significant advances have been achieved in DNA sequencing technologies, a positive identification of BWAs in environmental samples must be made through the ability of cells to form colony‐forming units upon culturing. Inability to revive the cells between collection and analysis renders such studies inconclusive. Commercial kits designed to preserve the viability of pathogens contained within clinical samples are available, but many of them have not been examined for their ability to preserve samples containing suspected BWAs. The study was initiated to examine the applicability of commercial solutions aiding in retention of Y. pestis viability in samples stored under nonpermissive temperatures, that is, 40 and 37°C. While none of the tested solutions sustained cell viability at 40°C, the results show five out of 17 tested preservatives were capable of supporting viability of Y. pestis at 37°C.

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Laboratory Maintenance and Characterization of Yersinia pestis

Cited by 8 publications

References 19 publications

Biosafety Level 2 Model of Pneumonic Plague and Protection Studies with F1 and Psa

Biosafety Level 2 Model of Pneumonic Plague and Protection Studies with F1 and Psa

Vulnerabilities in Yersinia pestis caf Operon Are Unveiled by a Salmonella Vector

Investigations into Enhancing Yersinia pestis Cells Viability following Environmental Sampling for Forensic Analysis

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