Abstract

I plan to study the pathogenesis of Salmonella infections using classical and molecular genetics. My long term goal is the development of a live Salmonella vaccine that can be used as a carrier to deliver heterologous antigens to the immune system. Phase I of this grant includes formal coursework in basic science necessary to study pathogenic bacteria and develop live attenuated bacterial vaccines. Phase I research training will be spent in the laboratory of Dr. John Mekalanos studying the molecular pathogenesis of Salmonella infections. Our preliminary investigations demonstrate that Salmonella typhimurium mutants deficient in the gene regulating acid phosphatase production (phoP) have reduced virulence in a mouse model of typhoid fever. We plan to investigate the cause of this reduced virulence. We also plan to identify S. typhimurium genes important to virulence by the use of several techniques including transposon mutagenesis to create gene translational fusions to E. coli alkaline phosphatase. We also plan to develop a vector delivery system for rapid and stable insertion of genes encoding heterologous antigens on the Salmonella chromosome.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Physician Scientist Award (K11)
Project #
5K11AI000917-05
Application #
3085283
Study Section
Microbiology and Infectious Diseases B Subcommittee (MID)
Project Start
1989-04-01
Project End
1994-03-31
Budget Start
1993-04-01
Budget End
1994-03-31
Support Year
5
Fiscal Year
1993
Total Cost
Indirect Cost

  Institution

Name
Massachusetts General Hospital
Department
Type
DUNS #
City
Boston
State
MA
Country
United States
Zip Code
02199

  Publications

Alpuche-Aranda, C M; Racoosin, E L; Swanson, J A et al. (1994) Salmonella stimulate macrophage macropinocytosis and persist within spacious phagosomes. J Exp Med 179:601-8
Miller, S I; Loomis, W P; Alpuche-Aranda, C et al. (1993) The PhoP virulence regulon and live oral Salmonella vaccines. Vaccine 11:122-5
Behlau, I; Miller, S I (1993) A PhoP-repressed gene promotes Salmonella typhimurium invasion of epithelial cells. J Bacteriol 175:4475-84
Ouellette, A J; Miller, S I; Henschen, A H et al. (1992) Purification and primary structure of murine cryptdin-1, a Paneth cell defensin. FEBS Lett 304:146-8
Alpuche Aranda, C M; Swanson, J A; Loomis, W P et al. (1992) Salmonella typhimurium activates virulence gene transcription within acidified macrophage phagosomes. Proc Natl Acad Sci U S A 89:10079-83
Miller, V L; Beer, K B; Loomis, W P et al. (1992) An unusual pagC::TnphoA mutation leads to an invasion- and virulence-defective phenotype in Salmonellae. Infect Immun 60:3763-70
Pulkkinen, W S; Miller, S I (1991) A Salmonella typhimurium virulence protein is similar to a Yersinia enterocolitica invasion protein and a bacteriophage lambda outer membrane protein. J Bacteriol 173:86-93