? We are proposing a translational investigation, including clinical as well as basic scientists, to test the hypothesis that specific Pseudomonas virulence products determine whether lung infection occurs. The results from the proposed experiments will be used to develop diagnostic tools that can identify virulent Pseudomonas in patients. The results will also identify critical bacterial gene products involved in lung injury. The present application carefully dissects associations between bacterial virulence and lung injury so that future mechanistic investigations in patients can be proposed. Without the data from the proposed investigations, we do not have the information to proceed. We already have Pseudomonas strains obtained from a cohort of intubated, mechanically ventilated patients from another grant.
Our first aim i s to perform a genomic analysis of Pseudomonas strains obtained from 2 distinct patient populations; intubated, ventilated patients with signs of lung infections that have disseminated and intubated, ventilated patients who have no sign of disease. Using microarray analysis, we will determine the significantly different genes between the Pseudomonas strains found in these two patient groups and produce consensus primers to these genes. We will then utilize these consensus primers in a prospective investigation of 300 intubated, ventilated patients to confirm the prevalence of the genetic differences in colonized vs infected intubated, ventilated patients.
The second aim i s to compare the in vivo virulence gene expression of Pseudomonas strains obtained from colonized patients to the expression profile of the strains obtained from patients who have Pseudomonas lung infections Analysis of bacterial gene expression in vivo will be done using real-time PCR. The results of all these experiments will allow us to prove (or disprove) that there are different genes or different patterns of toxin gene expression between the Pseudomonas strains infecting patients and those colonizing patients. In the final aim, we will evaluate the interaction of type III toxin genes in vivo By utilizing an isogenic bacterial strain (PA103) that has each of the toxin genes added alone or in combination, we will be able to define the effects of each of the type III toxins as well as other genes we find of significance in the first two aims. ? ?

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL069809-03
Application #
6994434
Study Section
Surgery, Anesthesiology and Trauma Study Section (SAT)
Program Officer
Harabin, Andrea L
Project Start
2003-12-11
Project End
2007-11-30
Budget Start
2005-12-01
Budget End
2006-11-30
Support Year
3
Fiscal Year
2006
Total Cost
$374,163
Indirect Cost
Name
University of California San Francisco
Department
Anesthesiology
Type
Schools of Medicine
DUNS #
094878337
City
San Francisco
State
CA
Country
United States
Zip Code
94143
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Fricks-Lima, J; Hendrickson, C M; Allgaier, M et al. (2011) Differences in biofilm formation and antimicrobial resistance of Pseudomonas aeruginosa isolated from airways of mechanically ventilated patients and cystic fibrosis patients. Int J Antimicrob Agents 37:309-15
Srinivasan, Ramya; Song, Yuanlin; Wiener-Kronish, Jeanine et al. (2011) Plasminogen activation inhibitor concentrations in bronchoalveolar lavage fluid distinguishes ventilator-associated pneumonia from colonization in mechanically ventilated pediatric patients. Pediatr Crit Care Med 12:21-7
Singh, G; Wu, B; Baek, M S et al. (2010) Secretion of Pseudomonas aeruginosa type III cytotoxins is dependent on pseudomonas quinolone signal concentration. Microb Pathog 49:196-203
Lynch, Susan V; Flanagan, Judith L; Sawa, Teiji et al. (2010) Polymorphisms in the Pseudomonas aeruginosa type III secretion protein, PcrV - implications for anti-PcrV immunotherapy. Microb Pathog 48:197-204
Yang, Katherine; Zhuo, Hanjing; Guglielmo, B Joseph et al. (2009) Multidrug-resistant Pseudomonas aeruginosa ventilator-associated pneumonia: the role of endotracheal aspirate surveillance cultures. Ann Pharmacother 43:28-35
El-Solh, Ali A; Amsterdam, Daniel; Alhajhusain, Ahmad et al. (2009) Matrix metalloproteases in bronchoalveolar lavage fluid of patients with type III Pseudomonas aeruginosa pneumonia. J Infect 59:49-55
Wiener-Kronish, Jeanine P; Dorr, Henry Isaiah (2008) Ventilator-associated pneumonia: problems with diagnosis and therapy. Best Pract Res Clin Anaesthesiol 22:437-49
Lynch, Susan V; Wiener-Kronish, Jeanine P (2008) Novel strategies to combat bacterial virulence. Curr Opin Crit Care 14:593-9
El Solh, Ali A; Akinnusi, Morohunfolu E; Wiener-Kronish, Jeanine P et al. (2008) Persistent infection with Pseudomonas aeruginosa in ventilator-associated pneumonia. Am J Respir Crit Care Med 178:513-9

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