Molecular Pathogenesis of Rickettsioses and Development of Novel Anti-Rickettsia Treatment by Comginatorial Peptide-Based Libraries
Abstract
The purpose of this study is to utilize adaptein libraries coded within pantropic retroviral vectors that confer protection against rickettsial pathogens and to study the molecular pathogenesis of rickettsioses. The following specific aims were proposed: 1) To establish heterogeneous cell populations, with each cell expressing 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, 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 third year of the project, rickettsial challenges performed with the transfected rat derived microvascular endothelial cells and the human brain primary microvascular endothelial cells were somewhat disappointing. Expansion of the "resistant colonies" was not possible. Two other human microvascular endothelial cell lines were acquired (cerebral and dermal). These cell lines are far more susceptible to rickettsial infection than their predecessors. We have transfected successfully both cell lines with the adaptein-containing retroviral vectors and "resistant colonies" were obtained after three consecutive challenges. However, expansion of the colonies continues to be elusive. Supernatants of infected monolayers were shown to affect the endothelial monolayer permeability dramatically, suggesting the presence of soluble factors, yet to be characterized, responsible for such changes. Excellent progress continues to be made on the development of an in vitro model for the study of microvascular permeability using human derived microvascular cells of cerebral and dermal origin.
Document Details
- Document Type
- Technical Report
- Publication Date
- Feb 01, 2005
- Accession Number
- ADA442166
Entities
People
- David H. Walker
- Juan P. Olano
Organizations
- University of Texas at Austin