Date of Award
2007
Degree Type
Thesis
Degree Name
Master of Science
Program
Microbiology and Immunology
Supervisor
Dr. Alexandra Lucas
Abstract
: The myxomaviral serpin, Serp-1, is a highly effective anti-inflammatory agent. The mechanism by which it binds to and inhibits monocyte activation remains undefined. Serp-1 was tested for association and internalization into THP-1 monocytes and Jurkat T- cells using spectrofluorometry. FACS analysis and a range of activating agents in the presence and absence of blocking antibodies to cell surface proteins was performed. Immunoprecipitation of uPA pretreated THP-1 monocyte membrane proteins was performed, but without an untreated or silver-stained control. Ex vivo treatment of a murine peritoneal exudate was attempted, but without a negative control. Serp-1 associated with both cell types and was internalized, but isolation of the nuclear envelope was not confirmed. Association was partially inihibited in THP-1 monocytes with uPAR blocking antibodies. Spectrofluorometry revealed activation of THP-1 monocytes with uPA resulted in modest increases in Serp-1 binding compared to albumin control (p< 0.0017). Immunoprecipitation with uPAR pulled down Serp-1, filamin-B, CDllb and CD61. Association with Jurkat T-cells is CD1lb/uPAR dependent.
Recommended Citation
Richardson, Jakob Gintaras, "ELUCIDATION OF THE ANTI-INFLAMMATORY VIRAL SERPIN SERP-l’S CELLULAR ASSOCIATION AND MODULATION OF MONOCYTE ADHESION THROUGH DIFFERENTIAL EXPRESSION OF THE ACTIN BINDING PROTEIN FILAMIN-B" (2007). Digitized Theses. 5125.
https://ir.lib.uwo.ca/digitizedtheses/5125