Abstract

The central goal of this project is to understand the role of cellular immunity in infections caused by LPS-smooth strains of P. aeruginosa. These strains are significant pathogens for hospitalized patients and wearers of extended-use contact lenses. Recent evidence suggests that during the course of lung and eye infections, P. aeruginosa enters epithelial cells via the cystic fibrosis transmembrane conductance regulator. To study the immune response to the intracellular base of infection, live, attenuated P. aeruginosa strains having an unmarked deletion of the aroA gene have been constructed. Preliminary studies reveal that intranasal (IN) immunization with one such mutant protects against keratitis or lethal pneumonia in murine models. Splenic T cells from immunized mice can kill intracellular P. aeruginosa. Interestingly, passive transfer of antiserum is more protective against corneal infections than lung infections. The candidate will further define the cellular immunity to aroA deletion mutants of P. aeruginosa and test the following hypothesis: The protective efficacy of IN immunization with live, attenuated P. aeruginosa vaccine strains requires both humoral and cellular immune effecters, and the relative contribution of each effecter towards protection is different in different sites of infection.
In Aim 1, the cellular immune effecters elicited by aroA deletants of P. aeruginosa will be delineated using immunomagnetic depletion coupled with assays for intracellular bacterial killing, T cell proliferation, and cytolysis.
In Aim 2, P. aeruginosa specific T cell clones from IN-immunized mice will be derived and characterized. The candidate seeks an intensive, formal, mentored training to provide the necessary intellectual and technical tools to achieve independence as a scientist. As a specialist in pediatric critical care and infectious diseases, his long-term goal is to develop effective immune-based interventions to prevent or ameliorate the consequences of P. aeruginosa infections.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Clinical Investigator Award (CIA) (K08)
Project #
5K08AI050036-04
Application #
6755956
Study Section
Microbiology and Infectious Diseases B Subcommittee (MID)
Program Officer
Korpela, Jukka K
Project Start
2001-07-01
Project End
2006-06-30
Budget Start
2004-07-01
Budget End
2005-06-30
Support Year
4
Fiscal Year
2004
Total Cost
$128,520
Indirect Cost

  Institution

Name
Brigham and Women's Hospital
Department
Type
DUNS #
030811269
City
Boston
State
MA
Country
United States
Zip Code
02115

  Publications

Long, Kelly R; Lomonosova, Elena; Li, Qilan et al. (2018) Efficacy of hepatitis B virus ribonuclease H inhibitors, a new class of replication antagonists, in FRG human liver chimeric mice. Antiviral Res 149:41-47
Sakala, Isaac G; Eickhoff, Christopher S; Blazevic, Azra et al. (2013) Dipterinyl calcium pentahydrate inhibits intracellular mycobacterial growth in human monocytes via the C-C chemokine MIP-1? and nitric oxide. Infect Immun 81:1974-83
Morrey, John D; Motter, Neil E; Chang, Stella et al. (2011) Breaking B and T cell tolerance using cationic lipid--DNA complexes (CLDC) as a vaccine adjuvant with hepatitis B virus (HBV) surface antigen in transgenic mice expressing HBV. Antiviral Res 90:227-30
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
Yu, Wenquan; Goddard, Cally; Clearfield, Elizabeth et al. (2011) Design, synthesis, and biological evaluation of triazolo-pyrimidine derivatives as novel inhibitors of hepatitis B virus surface antigen (HBsAg) secretion. J Med Chem 54:5660-70
Moheno, Phillip; Morrey, John; Fuchs, Dietmar (2010) Effect of dipterinyl calcium pentahydrate on hepatitis B virus replication in transgenic mice. J Transl Med 8:32
Morrey, John D; Siddharthan, Venkatraman; Wang, Hong et al. (2010) Neurological suppression of diaphragm electromyographs in hamsters infected with West Nile virus. J Neurovirol 16:318-29
Kamei, Akinobu; Koh, Andrew Y; Gadjeva, Mihaela et al. (2010) Analysis of acquisition of Pseudomonas aeruginosa gastrointestinal mucosal colonization and horizontal transmission in a murine model. J Infect Dis 201:71-80
Coughlin, John E; Padmanabhan, Seetharamaiyer; Zhang, Guangrong et al. (2010) Orally bioavailable anti-HBV dinucleotide acyloxyalkyl prodrugs. Bioorg Med Chem Lett 20:1783-6
Morrey, John D; Korba, Brent E; Beadle, James R et al. (2009) Alkoxyalkyl esters of 9-(s)-(3-hydroxy-2-phosphonomethoxypropyl) adenine are potent and selective inhibitors of hepatitis B virus (HBV) replication in vitro and in HBV transgenic mice in vivo. Antimicrob Agents Chemother 53:2865-70

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