Bacterial polysaccharides are excellent targets for vaccine development as they are surface exposed and often the most protective antigens expressed on a pathogen. However, purified polysaccharides are generally poor immunogens, particularly if their immunogenicity is not enhanced by conjugation to a protein. Such polysaccharide-protein conjugate vaccines are often complicated and expensive to produce and difficult to deliver. In order to demonstrate the potential of a novel approach to utilize polysaccharides as vaccine candidates, we have expressed the O antigen portion of Pseudomonas aeruginosa lipopolysaccharide (IPS) on an attenuated strain of Salmonella typhimurium. We compared different routes of immunization to protect against acute P. aeruginosa pneumonia in a mouse model system and found that intranasal administration of the vaccine provides better protection than either oral or intraperitoneal delivery. However, we do not know whether it is the location, level, and/or type of antibody response that is responsible for this protection. We have also shown that intranasal delivery of this vaccine can protect mice against P. aeruginosa infections after corneal trauma or after burns, but do not know the optimal route of immunization to induce immunity at these different sites, nor do we know the effect of host factors such as cystic fibrosis on modulating the immunity induced by the attenuated recombinant S. typhimurium. Also unclear is whether such vaccines can alleviate problems associated with polysaccharide-protein conjugate vaccines including subtype specificity and subtype inhibition, and the exclusion of acid-liable O antigens.
The specific aims of this grant are to (1) determine the basis for protective immunity to P. aeruginosa acute pneumonia mediated by the attenuated recombinant S. typhimurium O antigen-based vaccine, (2) determine the optimal route of immunization to protect against P. aeruginosa infections after corneal injury or burns, (3) determine whether the vaccine can protect CF mice from P. aeruginosa lung infections, and (4) test additional O antigen-based vaccines for polysaccharide expression, immunogenicity, and protection. Overall, the long-term goal of this project is to understand the basis of protection against these varied infections, which in turn will allow us to develop rational strategies for vaccine development against this important opportunistic pathogen. We anticipate that this knowledge will also be applicable to other pathogens.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
3R01AI068112-05S1
Application #
8147892
Study Section
Vaccines Against Microbial Diseases (VMD)
Program Officer
Taylor, Christopher E,
Project Start
2006-06-01
Project End
2012-05-31
Budget Start
2010-09-30
Budget End
2012-05-31
Support Year
5
Fiscal Year
2010
Total Cost
$134,702
Indirect Cost
Name
University of Virginia
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
065391526
City
Charlottesville
State
VA
Country
United States
Zip Code
22904
Damron, F Heath; McKenney, Elizabeth S; Barbier, Mariette et al. (2013) Construction of mobilizable mini-Tn7 vectors for bioluminescent detection of gram-negative bacteria and single-copy promoter lux reporter analysis. Appl Environ Microbiol 79:4149-53
Damron, F Heath; Barbier, Mariette; McKenney, Elizabeth S et al. (2013) Genes required for and effects of alginate overproduction induced by growth of Pseudomonas aeruginosa on Pseudomonas isolation agar supplemented with ammonium metavanadate. J Bacteriol 195:4020-36
Damron, F Heath; Goldberg, Joanna B (2012) Proteolytic regulation of alginate overproduction in Pseudomonas aeruginosa. Mol Microbiol 84:595-607
Damron, F Heath; Owings, Joshua P; Okkotsu, Yuta et al. (2012) Analysis of the Pseudomonas aeruginosa regulon controlled by the sensor kinase KinB and sigma factor RpoN. J Bacteriol 194:1317-30
Kamei, Akinobu; Coutinho-Sledge, Yamara S; Goldberg, Joanna B et al. (2011) Mucosal vaccination with a multivalent, live-attenuated vaccine induces multifactorial immunity against Pseudomonas aeruginosa acute lung infection. Infect Immun 79:1289-99
Ivanov, Ivan E; Kintz, Erica N; Porter, Laura A et al. (2011) Relating the physical properties of Pseudomonas aeruginosa lipopolysaccharides to virulence by atomic force microscopy. J Bacteriol 193:1259-66
Damron, F Heath; Davis Jr, Michael R; Withers, T Ryan et al. (2011) Vanadate and triclosan synergistically induce alginate production by Pseudomonas aeruginosa strain PAO1. Mol Microbiol 81:554-70
Kintz, Erica N; Goldberg, Joanna B (2011) Site-directed mutagenesis reveals key residue for O antigen chain length regulation and protein stability in Pseudomonas aeruginosa Wzz2. J Biol Chem 286:44277-84
Goldberg, Joanna B (2010) Why is Pseudomonas aeruginosa a pathogen? F1000 Biol Rep 2:
Priebe, Gregory P; Walsh, Rebecca L; Cederroth, Terra A et al. (2008) IL-17 is a critical component of vaccine-induced protection against lung infection by lipopolysaccharide-heterologous strains of Pseudomonas aeruginosa. J Immunol 181:4965-75

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