Molecular Pathogenesis of Rickettsioses and Development of Novel Anti-Rickettsia Treatment by Combinatorial Peptide-based Libraries
Abstract
The purpose of this study is to utilize adaptein libraries coded within pantropic retroviral vectors the confer protection against rickettsial pathogens and to study themolecular pathogenesis of rickettsioses. The following specific aims were proposed: 1) To establish heterogeneous cell populations, with each cell expression a unique member of a complex combinatorial peptide-based (e.g., adaptein) library and challenge with R. prowazekii, R. rickettsii, and 0. tsutsugamushi; 2) To determine the role of NF-kB, cytokines (TNF-a, IFN-g, RANTES), ROS, and NO in intracellular killing of rickettsia- infected monolayers containing adapteins and 3) To characterize signal transduction pathways modulating the cytoskeletal events responsible for the increased vascular permeability. During the second year of the project, we were ale to produce and optimize production of adaptein libraries containing 6-, 12-, and 18-mer peptides and EGFP in he retroviral vector. Successful transfection of rat derived brain microvascular endothelial cells and human brain primary microvascular endothelial cells with the recombinant retroviruses was achieved. The results of rickettsial challenges of adaptein-producing RBE4 cells are very encouraging. Successful transfection of RBE4 cells with the pNF-kB- d2EGFP plasmid and demonstration of NF-kB activation after rickettsial infection were also achieved. Great progress was made in development of in vitro models of endothelial barrier using rat-derived and human derived brain microvascular endothelial cells. Demonstration of the role of cytokines in modulation of microvascular permeability and possible role of ROS, calcium and NO in endothelial permeability was also demonstrated.
Document Details
- Document Type
- Technical Report
- Publication Date
- Feb 01, 2004
- Accession Number
- ADA426090
Entities
People
- David H. Walker
Organizations
- University of Texas at Austin