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  4. Bacterial Vaccine Safety: Biomarkers of Virulence and Attenuation in Bordetella Pertussis (Whooping Cough) and Anthrax Bacteria
  1. Science & Research (Biologics)

Bacterial Vaccine Safety: Biomarkers of Virulence and Attenuation in Bordetella Pertussis (Whooping Cough) and Anthrax Bacteria

Principal Investigator: E. Scott Stibitz, PhD
Office / Division / Lab: OVRR / DBPAP / LESTD

General Overview

Our laboratory is studying disease-causing bacteria for which new vaccines and novel therapeutic approaches are being developed.

We are developing new ways to study respiratory infections caused by the bacterium Bordetella pertussis (whooping cough) and the mechanisms by which it causes severe disease. Although a vaccine for this organism exists, the number of whooping cough cases in the US continues to rise, making this research vital to ultimate control.

We also support the development and evaluation of vaccines against Staphylococcus aureus, which causes many infections in the US each year, some of which are extremely serious (e.g., "flesh-eating" bacteria that can cause death in a matter of hours). S. aureus also causes infections of implanted devices such as pacemakers and joint replacements that can be very difficult to treat without removing the device. Currently no vaccine for this disease exists and development of new vaccines against this organism is especially critical due to problems with antibiotic resistance typified by MRSA (methicillin-resistant Staphylococcus aureus), and the new strains of community-acquired MRSA (CA-MRSA).

We also study live biotherapeutic products (i.e., probiotics--live microorganisms that provide health benefits to an individual when consumed in appropriate quantities) to help treat and prevent bacterial infections and other conditions. The investigation of the full potential of these promising products is currently being hindered by difficulties in evaluation of the manufacturing process, purity, safety and quality of these products. We seek to solve this problem by developing better tests for the purity and safety of probiotics, thereby allowing their further testing and development in the clinic.

Diarrheal diseases are also targetted. We are investigating the use of the live attenuated Salmonella strain Ty21a as a platform for eliciting protective immune responses against other serious diseases such as Shigellosis. We are also examining ways to more fully understand some of the as yet uncharacterized genetic bases of attenuation in Ty21a as well as to confer advantageous traits such as acid resistance. We are also seeking to understand why some bacterial pathogens, such as Shigella can cause arthritis and are testing novel approaches to killed Shigella vaccines in order to control this deadly disease.

Another major worldwide health challenge and emerging problem in the US - tuberculosis - is in a class by itself in terms of its difficulty to study, its overall importance, its resistance to date to vaccine interventions, and the threat from antibiotic resistance. Our lab is working to develop and execute new ways to assess the usefullness of vaccine candidates, including the development of animal models and of in vitro tests to measure host responses.

Malaria, while not a bacterial disease, in one which also has a huge worldwide burden. Recent exciting research has brought us much closer to having a useful anti-malaria vaccine. Our lab is studying biomarkers for severe malaria in an effort to both better understand the variables that can lead to tragic outcomes as well as to better assess how current vaccine candidates control disease and infection.

Scientific Overview

Our laboratory takes a genetic approach to studies of bacterial pathogenesis and vaccine development.

In our studies of Bordetella pertussis we introduce specific mutations that affect the expression and regulation of virulence factors, such as toxins and adhesins. We then test the effects of these mutations in a mouse model of respiratory infection that uses bioluminescent B. pertussis to track infection without harming the test animal. This approach has several advantages over older methods that involve sacrificing a group of test animals at each time point, and reduces dramatically the number of animals that must be used.

Our approach to studying Bacillus anthracis pathogenesis has also been largely genetic. We developed and have recently improved, new genetic tools to introduce specific mutations into this organism. These tools, which are now used widely around the world, are significantly easier and more powerful than previous such techniques. They also allow the consecutive deletion or mutation of a unlimited number of genes. This approach has been critical to our studies of anthrax vaccine stability. For example, it has enabled us to create improved strains of the organism used to produce protective antigen (part of the anthrax toxin). These strains lack a number of secreted proteases (currently up to 16 separate deletions). This characteristic increases the production of protective antigen and leads to a purified product that has increased stability.

Our new program in Staphylococcus aureus will follow a similar path, synergizing our genetic capabilities with other OVRR researchers to evaluate new vaccine candidates and identify new candidates for virulence factors and vaccine antigens. We are also examining the utility of a novel approach - bacteriophage therapy - to deal with antibiotic resistant S. aureus.

We are using bacteriophages to develop improved tests for detecting pathogens in probiotic products. We plan to exploit the exquisite specificity of these bacterial viruses to specifically kill product organisms, thereby increasing the sensitivity of detection of extraneous and potentially harmful bacterial pathogens, such as Salmonella and Shigella. Such tests are necessary to ensure the safety of these preparations, a factor that is especially important when tested in clinical trials in very sick or otherwise compromised patients.

Another application of bacteriophages is to actually treat bacterial diseases. Our lab is initiating a project to set up an animal model to examine the usefullness of phage in host decolonization of Staphylococcus aureus. If successful, this could have a major impact on carriage in hospitals and significantly improve the situation there where dangerous, highly antibiotic strains, MRSA, have a high toll.


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    Conformational change of the Bordetella response regulator BvgA accompanies its activation of the B. pertussis virulence gene fhaB.
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    Bacteriophage-antibiotic combination therapy for multidrug-resistant Pseudomonas aeruginosa: in vitro synergy testing.
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    RNase III and RNase E influence posttranscriptional regulatory networks involved in virulence factor production, metabolism, and regulatory RNA processing in Bordetella pertussis.
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    Four single-basepair mutations in the ptx promoter of Bordetella bronchiseptica are sufficient to activate the expression of pertussis toxin.
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    Multiple weak interactions between BvgA~P and ptx promoter DNA strongly activate transcription of pertussis toxin genes in Bordetella pertussis.
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    The BvgASR virulence regulon of Bordetella pertussis.
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