The long-term objectives and specific aims of this application are to understand the pathogenesis of Rickettsia rickettsii, and other members of the spotted fever group of rickettsiae by better studying the processes involved in causing cell injury. The investigators are interested in the interaction of R. rickettsii (Rocky Mountain spotted fever) with the endothelial cell, the target cell in human infection, and the mechanism(s) of cell injury caused by this obligate intracellular bacterium. It has been determined by the Principal Investigator's laboratory that cell injury appears to be initiated by reactive oxygen species produced during internalization and intracellular rickettsial growth. With an interest in abrogating endothelial cell injury, they have incorporated antioxidants into infected cell culture systems. One antioxidant, (alpha-lipoic acid, was able to effectively reverse the leading predictors of cell injury due to reactive oxygen species and to substantially prolong the viability of infected cell cultures. Studies to date have been carried out both in human umbilical vein endothelial cells and the permanent human endothelial cell line, EA.hy926. The investigators will continue to carry out experiments in these cells to characterize changes in the cytoskeleton of endothelial cells following R. rickettsii infection and to evaluate the link between these changes and increases in endothelial permeability. They will determine whether R. rickettsii-induced oxidative injury is responsible for the alterations in the actin pools and whether these changes can be prevented (controlled) by the use of antioxidants. They believe that it is also important to test our in vitro observations on oxidant-mediated injury in a relevant animal model to determine whether oxidant injury occurs and if antioxidants are effective in altering the course of disease. The most suitable model for these studies is the Rickettsia conorii-C3H/HeN mouse model which has been extensively studied by Walker and his colleagues. R. conorii is closely related to R. rickettsii, causes Mediterranean spotted fever, a syndrome clinically similar to RMSF, and as they have demonstrated, results in the same dramatic morphological manifestations of cell injury as R. rickettsii in human endothelial cells. These preliminary in vitro studies and those in the mouse model, indicate that this rickettsia is also able to cause oxidative stress. They propose to test the hypothesis that oxidant injury contributes to the pathologic changes within the animal. They will determine whether oxidative injury correlates with the local presence of rickettsiae within particular organs and tissues and whether the antioxidant, alpha-lipoic acid, can ameliorate oxidative injury as they have shown in vitro.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI017416-16
Application #
6169392
Study Section
Bacteriology and Mycology Subcommittee 2 (BM)
Program Officer
Baker, Phillip J
Project Start
1980-12-01
Project End
2003-07-31
Budget Start
2000-08-01
Budget End
2001-07-31
Support Year
16
Fiscal Year
2000
Total Cost
$333,093
Indirect Cost
Name
University of Maryland Baltimore
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
003255213
City
Baltimore
State
MD
Country
United States
Zip Code
21201
Rydkina, Elena; Silverman, David J; Sahni, Sanjeev K (2005) Activation of p38 stress-activated protein kinase during Rickettsia rickettsii infection of human endothelial cells: role in the induction of chemokine response. Cell Microbiol 7:1519-30
Rydkina, Elena; Sahni, Sanjeev K; Santucci, Lisa A et al. (2004) Selective modulation of antioxidant enzyme activities in host tissues during Rickettsia conorii infection. Microb Pathog 36:293-301
Joshi, Suresh G; Francis, Charles W; Silverman, David J et al. (2004) NF-kappaB activation suppresses host cell apoptosis during Rickettsia rickettsii infection via regulatory effects on intracellular localization or levels of apoptogenic and anti-apoptotic proteins. FEMS Microbiol Lett 234:333-41
Eremeeva, Marina E; Klemt, Ryan M; Santucci-Domotor, Lisa A et al. (2003) Genetic analysis of isolates of Rickettsia rickettsii that differ in virulence. Ann N Y Acad Sci 990:717-22
Eremeeva, Marina E; Liang, Zhongxing; Paddock, Christopher et al. (2003) Rickettsia rickettsii infection in the pine vole, Microtus pinetorum: kinetics of infection and quantitation of antioxidant enzyme gene expression by RT-PCR. Ann N Y Acad Sci 990:468-73
Eremeeva, Marina E; Dasch, Gregory A; Silverman, David J (2003) Evaluation of a PCR assay for quantitation of Rickettsia rickettsii and closely related spotted fever group rickettsiae. J Clin Microbiol 41:5466-72
Sahni, Sanjeev K; Rydkina, Elena; Joshi, Suresh G et al. (2003) Interactions of Rickettsia rickettsii with endothelial nuclear factor-kappaB in a ""cell-free"" system. Ann N Y Acad Sci 990:635-41
Joshi, Suresh G; Francis, Charles W; Silverman, David J et al. (2003) Nuclear factor kappa B protects against host cell apoptosis during Rickettsia rickettsii infection by inhibiting activation of apical and effector caspases and maintaining mitochondrial integrity. Infect Immun 71:4127-36
Rydkina, Elena; Sahni, Abha; Silverman, David J et al. (2002) Rickettsia rickettsii infection of cultured human endothelial cells induces heme oxygenase 1 expression. Infect Immun 70:4045-52
Eremeeva, M E; Dasch, G A; Silverman, D J (2001) Quantitative analyses of variations in the injury of endothelial cells elicited by 11 isolates of Rickettsia rickettsii. Clin Diagn Lab Immunol 8:788-96

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