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

ABSTRACT 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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Characterization of an F1 Deletion Mutant of Yersinia pestis CO92, Pathogenic Role of F1 Antigen in Bubonic and Pneumonic Plague, and Evaluation of Sensitivity and Specificity of F1 Antigen Capture-Based Dipsticks

Journal of Clinical Microbiology ◽

10.1128/jcm.00064-11 ◽

2011 ◽

Vol 49 (5) ◽

pp. 1708-1715 ◽

Cited By ~ 28

Author(s):

J. Sha ◽

J. J. Endsley ◽

M. L. Kirtley ◽

S. M. Foltz ◽

M. B. Huante ◽

...

Keyword(s):

Yersinia Pestis ◽

Sensitivity And Specificity ◽

Deletion Mutant ◽

Pathogenic Role ◽

Pneumonic Plague ◽

Antigen Capture ◽

F1 Antigen

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The Yersinia pestis GTPase BipA Promotes Pathogenesis of Primary Pneumonic Plague

Infection and Immunity ◽

10.1128/iai.00673-20 ◽

2020 ◽

Author(s):

Samantha D. Crane ◽

Srijon K. Banerjee ◽

Kara R. Eichelberger ◽

Richard C. Kurten ◽

William E. Goldman ◽

...

Keyword(s):

Yersinia Pestis ◽

Virulence Factors ◽

Virulence Factor ◽

Abrupt Onset ◽

Bacterial Killing ◽

Pneumonic Plague ◽

Distinct Disease ◽

Bacterial Replication ◽

Two Phases ◽

Type 3

Yersinia pestis is a highly virulent pathogen and the causative agent of bubonic, septicemic, and pneumonic plague. Primary pneumonic plague caused by inhalation of respiratory droplets contaminated with Y. pestis is nearly 100% lethal within 4-7 days without antibiotic intervention. Pneumonic plague progresses in two phases, beginning with extensive bacterial replication in the lung with minimal host responsiveness, followed by the abrupt onset of a lethal pro-inflammatory response. The precise mechanisms by which Y. pestis is able to colonize the lung and survive two very distinct disease phases remain largely unknown. To date, a handful of bacterial virulence factors including the Ysc type 3 secretion system are known to contribute to the pathogenesis of primary pneumonic plague. The bacterial GTPase BipA has been shown to regulate expression of virulence factors in a number of Gram-negative bacteria including Pseudomonas aeruginosa, Escherichia coli, and Salmonella enterica serovar Typhi. However, the role of BipA in Y. pestis has yet to be investigated. Here, we show that BipA is a Y. pestis virulence factor that promotes defense against early neutrophil-mediated bacterial killing in the lung. This work identifies a novel Y. pestis virulence factor, and highlights the importance of early bacterial/neutrophil interactions in the lung during primary pneumonic plague.

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Synergistic Protection of Mice against Plague with Monoclonal Antibodies Specific for the F1 and V Antigens of Yersinia pestis

Infection and Immunity ◽

10.1128/iai.71.4.2234-2238.2003 ◽

2003 ◽

Vol 71 (4) ◽

pp. 2234-2238 ◽

Cited By ~ 66

Author(s):

Jim Hill ◽

Catherine Copse ◽

Sophie Leary ◽

Anthony J. Stagg ◽

E. Diane Williamson ◽

...

Keyword(s):

Monoclonal Antibodies ◽

Yersinia Pestis ◽

Pneumonic Plague ◽

F1 Antigen ◽

V Antigen

ABSTRACT Monoclonal antibodies specific for Yersinia pestis V antigen and F1 antigen, administered singly or in combination, protected mice in models of bubonic and pneumonic plague. Antibodies showed synergy when administered prophylactically and as a therapy 48 h postinfection. Monoclonal antibodies therefore have potential as a treatment for plague.

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In Vivo Transcriptional Profiling of Yersinia pestis Reveals a Novel Bacterial Mediator of Pulmonary Inflammation

mBio ◽

10.1128/mbio.02302-14 ◽

2015 ◽

Vol 6 (1) ◽

Cited By ~ 14

Author(s):

Roger D. Pechous ◽

Christopher A. Broberg ◽

Nikolas M. Stasulli ◽

Virginia L. Miller ◽

William E. Goldman

Keyword(s):

Yersinia Pestis ◽

Inflammatory Responses ◽

Severe Pneumonia ◽

Sudden Onset ◽

Host Factors ◽

Respiratory Pathogen ◽

Bacterial Survival ◽

Pneumonic Plague ◽

Content Type

ABSTRACTInhalation ofYersinia pestisresults in primary pneumonic plague, a highly lethal and rapidly progressing necrotizing pneumonia. The disease begins with a period of extensive bacterial replication in the absence of disease symptoms, followed by the sudden onset of inflammatory responses that ultimately prove fatal. Very little is known about the bacterial and host factors that contribute to the rapid biphasic progression of pneumonic plague. In this work, we analyzed thein vivotranscription kinetics of 288 bacterial open reading frames previously shown by microarray analysis to be dynamically regulated in the lung. Using this approach combined with bacterial genetics, we were able to identify five Y. pestis genes that contribute to the development of pneumonic plague. Deletion of one of these genes,ybtX, did not alter bacterial survival but attenuated host inflammatory responses during late-stage disease. Deletion ofybtXin another lethal respiratory pathogen,Klebsiella pneumoniae, also resulted in diminished host inflammation during infection. Thus, ourin vivotranscriptional screen has identified an important inflammatory mediator that is common to two Gram-negative bacterial pathogens that cause severe pneumonia.IMPORTANCEYersinia pestis is responsible for at least three major pandemics, most notably the Black Death of the Middle Ages. Due to its pandemic potential, ease of dissemination by aerosolization, and a history of its weaponization, Y. pestis is categorized by the Centers for Disease Control and Prevention as a tier 1 select agent most likely to be used as a biological weapon. To date, there is no licensed vaccine against Y. pestis. Importantly, an early “silent” phase followed by the rapid onset of nondescript influenza-like symptoms makes timely treatment of pneumonic plague difficult. A more detailed understanding of the bacterial and host factors that contribute to pathogenesis is essential to understanding the progression of pneumonic plague and developing or enhancing treatment options.

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Inhibition of Neutrophil Primary Granule Release during Yersinia pestis Pulmonary Infection

mBio ◽

10.1128/mbio.02759-19 ◽

2019 ◽

Vol 10 (6) ◽

Cited By ~ 4

Author(s):

Kara R. Eichelberger ◽

Grant S. Jones ◽

William E. Goldman

Keyword(s):

Yersinia Pestis ◽

Critical Role ◽

Immune Defense ◽

Calcium Flux ◽

Human Neutrophils ◽

Pneumonic Plague ◽

Content Type ◽

Neutrophil Degranulation ◽

Primary Granule ◽

Granule Release

ABSTRACT Inhalation of Yersinia pestis causes primary pneumonic plague, the most severe manifestation of plague that is characterized by a dramatic neutrophil influx to the lungs. Neutrophils are ineffective during primary pneumonic plague, failing to control Y. pestis growth in the airways. However, the mechanisms by which Y. pestis resists neutrophil killing are incompletely understood. Here, we show that Y. pestis inhibits neutrophil degranulation, an important line of host innate immune defense. We observed that neutrophils from the lungs of mice infected intranasally with Y. pestis fail to release primary granules throughout the course of disease. Using a type III secretion system (T3SS) injection reporter strain, we determined that Y. pestis directly inhibits neutrophil granule release by a T3SS-dependent mechanism. Combinatorial mutant analysis revealed that a Y. pestis strain lacking both effectors YopE and YopH did not inhibit primary granule release and is killed by neutrophils both in vivo and in vitro. Similarly, Y. pestis strains injecting only YopE or YopH are able to inhibit the majority of primary granule release from human neutrophils. We determined that YopE and YopH block Rac2 activation and calcium flux, respectively, to inhibit neutrophil primary granule release in isolated human neutrophils. These results demonstrate that Y. pestis coordinates the inhibition of neutrophil primary granule release through the activities of two distinct effectors, and this inhibition promotes Y. pestis survival during primary pneumonic plague. IMPORTANCE Yersinia pestis is the causative agent of plague and is one of the deadliest human pathogens. The pneumonic form of Y. pestis infection has played a critical role in the severity of both historical and modern plague outbreaks, yet the host-pathogen interactions that govern the lethality of Yersinia pestis pulmonary infections are incompletely understood. Here, we report that Yersinia pestis inhibits neutrophil degranulation during infection, rendering neutrophils ineffective and allowing unrestricted growth of Y. pestis in the lungs. This coordinated inhibition of granule release not only demonstrates the pathogenic benefit of “silencing” lung neutrophils but also reveals specific host processes and pathways that could be manipulated to reduce the severity of primary pneumonic plague.

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Specific Detection of Yersinia pestis Based on Receptor Binding Proteins of Phages

Pathogens ◽

10.3390/pathogens9080611 ◽

2020 ◽

Vol 9 (8) ◽

pp. 611 ◽

Cited By ~ 1

Author(s):

Friederike Born ◽

Peter Braun ◽

Holger C. Scholz ◽

Gregor Grass

Keyword(s):

Yersinia Pestis ◽

Receptor Binding ◽

Fluorescent Proteins ◽

Diagnostic Methods ◽

Tail Fiber ◽

Yersinia Species ◽

Recombinant Receptor ◽

F1 Antigen ◽

Phage Receptor ◽

Bacterial Plaque

The highly pathogenic bacterium Yersinia pestis is the causative agent of plague, a notorious infectious zoonotic disease. When transmitted from person to person as pneumonic plague via droplets, Y. pestis is highly contagious and in most cases is fatal if left untreated. Thus, when plague is suspected, rapid diagnosis is crucial, as a serious course of the infection is only averted by early antibiotic therapy. The bacterium is easy to cultivate, accessible and has a high potential for nefarious use such as bioterrorism. Highly specific, rapid and easy-to-use confirmatory diagnostic methods are required to reliably identify the pathogen independently from PCR-based methods or F1 antigen-based immunological detection. Yersinia pestis specific phages such as L-413C and ΦA1122 are already used for detection of Y. pestis in bacterial plaque or biosensor assays. Here, we made use of the host specificities conferred by phage receptor binding (or tail fiber/spike) proteins (RBP) for developing a specific, fast and simple fluorescence-microscopy-based detection method for Y. pestis. Genes of putative RBP of phages L-413C (gpH) and ΦA1122 (gp17) were fused with those of fluorescent proteins and recombinant receptor-reporter fusion proteins were produced heterologously in Escherichia coli. When first tested on attenuated Y. pestis strain EV76, RBP-reporters bound to the bacterial cell surface. This assay could be completed within a few minutes using live or formaldehyde-inactivated cells. Specificity tests using cultures of closely related Yersinia species and several inactivated fully virulent Y. pestis strains exhibited high specificities of the RBP-reporters against Y. pestis. The L-413C RBP proved to be especially specific, as it only detected Y. pestis at all temperatures tested, whereas the RBP of ΦA1122 also bound to Y. pseudotuberculosis strains at 37 °C (but not at 28, 20 or 6 °C). Finally, the Y. pestis-specific capsule, produced when grown at 37 °C, significantly reduced binding of phage ΦA1122 RBP, whereas the capsule only slightly diminished binding of L-413C RBP.

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Flagellin Is an Effective Adjuvant for Immunization against Lethal Respiratory Challenge with Yersinia pestis

Infection and Immunity ◽

10.1128/iai.74.2.1113-1120.2006 ◽

2006 ◽

Vol 74 (2) ◽

pp. 1113-1120 ◽

Cited By ~ 188

Author(s):

Anna N. Honko ◽

Nammalwar Sriranganathan ◽

Cynthia J. Lees ◽

Steven B. Mizel

Keyword(s):

Yersinia Pestis ◽

Neutrophil Infiltration ◽

Specific Ige ◽

Innate Immune ◽

Igg Antibody ◽

Adjuvant Activity ◽

Necrosis Factor Alpha ◽

Potent Inducer ◽

Cytokines And Chemokines ◽

F1 Antigen

ABSTRACT Gram-negative flagellin, a Toll-like receptor 5 (TLR5) agonist, is a potent inducer of innate immune effectors such as cytokines and nitric oxide. In the lung, flagellin induces a localized and transient innate immune response characterized by neutrophil infiltration and the production of cytokines and chemokines. In view of the extraordinary potency of flagellin as an inducer of innate immunity and the contribution of innate responses to the development of adaptive immunity, we evaluated the efficacy of recombinant Salmonella flagellin as an adjuvant in an acellular plague vaccine. Mice immunized intranasally or intratracheally with the F1 antigen of Yersinia pestis and flagellin exhibited dramatic increases in anti-F1 plasma immunoglobulin G (IgG) titers that remained stable over time. In contrast, control mice had low or undetectable antibody responses. The IgG1/IgG2a ratio of antibody titers against F1 in immunized mice is consistent with a Th2 bias. However, no significant antigen-specific IgE production was detected. Interferons, tumor necrosis factor alpha, and interleukin-6 were not essential for the adjuvant effects of flagellin. Preexisting antiflagellin antibodies had no significant effect on the adjuvant activity of flagellin. Importantly, intranasal immunization with flagellin and the F1 antigen was protective against intranasal challenge with virulent Y. pestis CO92, with 93 to 100% survival of immunized mice. Lastly, vaccination of cynomolgus monkeys with flagellin and a fusion of the F1 and V antigens of Y. pestis induced a robust antigen-specific IgG antibody response.

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Yersinia pestis CO92ΔyopH Is a Potent Live, Attenuated Plague Vaccine

Clinical and Vaccine Immunology ◽

10.1128/cvi.00137-07 ◽

2007 ◽

Vol 14 (9) ◽

pp. 1235-1238 ◽

Cited By ~ 30

Author(s):

Sarah S. Bubeck ◽

Peter H. Dube

Keyword(s):

Yersinia Pestis ◽

Live Attenuated Vaccine ◽

Pneumonic Plague ◽

Degree Of Protection ◽

Attenuated Vaccine ◽

Respiratory Challenge ◽

High Degree ◽

Plague Vaccine

ABSTRACT An in-frame deletion of the yopH gene in Yersinia pestis CO92 attenuates virulence in both bubonic and pneumonic plague models. When it is used as a live, attenuated vaccine, CO92ΔyopH provides a high degree of protection from parental and respiratory challenge with Y. pestis CO92.

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A Replication-Defective Human Type 5 Adenovirus-Based Trivalent Vaccine Confers Complete Protection against Plague in Mice and Nonhuman Primates

Clinical and Vaccine Immunology ◽

10.1128/cvi.00150-16 ◽

2016 ◽

Vol 23 (7) ◽

pp. 586-600 ◽

Cited By ~ 10

Author(s):

Jian Sha ◽

Michelle L. Kirtley ◽

Curtis Klages ◽

Tatiana E. Erova ◽

Maxim Telepnev ◽

...

Keyword(s):

Fusion Gene ◽

The United States ◽

Pneumonic Plague ◽

Content Type ◽

Human Type ◽

Protective Antigens ◽

Type 3 Secretion System ◽

Needle Protein ◽

Trivalent Vaccine ◽

Type 3

Currently, no plague vaccine exists in the United States for human use. The capsular antigen (Caf1 or F1) and two type 3 secretion system (T3SS) components, the low-calcium-response V antigen (LcrV) and the needle protein YscF, represent protective antigens ofYersinia pestis. We used a replication-defective human type 5 adenovirus (Ad5) vector and constructed recombinant monovalent and trivalent vaccines (rAd5-LcrV and rAd5-YFV) that expressed either the codon-optimizedlcrVor the fusion gene designatedYFV(consisting ofycsF,caf1, andlcrV). Immunization of mice with the trivalent rAd5-YFV vaccine by either the intramuscular (i.m.) or the intranasal (i.n.) route provided protection superior to that with the monovalent rAd5-LcrV vaccine against bubonic and pneumonic plague when animals were challenged withY. pestisCO92. Preexisting adenoviral immunity did not diminish the protective response, and the protection was always higher when mice were administered one i.n. dose of the trivalent vaccine (priming) followed by a single i.m. booster dose of the purified YFV antigen. Immunization of cynomolgus macaques with the trivalent rAd5-YFV vaccine by the prime-boost strategy provided 100% protection against a stringent aerosol challenge dose of CO92 to animals that had preexisting adenoviral immunity. The vaccinated and challenged macaques had no signs of disease, and the invading pathogen rapidly cleared with no histopathological lesions. This is the first report showing the efficacy of an adenovirus-vectored trivalent vaccine against pneumonic plague in mouse and nonhuman primate (NHP) models.

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