Project 2

Project 2: Ischemia-Reperfusion Injury following Transplantation: implications for the modulation of CXCL8 and CXCL1 chemokine function

Injury following reperfusion of the ischemic organ, following transplantation is a major contributor to subsequent graft dysfunction. Furthermore, Post-translational modifications of chemokines, which are generated during inflammation, such as nitration or citrullination, can significantly affect their function.

BMB has characterised HL-60 neutrophil cell line and PMN for their chemokine receptor expression. Furthermore, she is studying the role of nitration on chemokine CXCL8 function. CXCL8 nitration could significantly decrease trans-endothelial neutrophil migration (P<0.05), possibly altering the chemokine receptor-binding or endothelial glycosaminoglycan-binding interaction. It has been shown through chemotaxis, and through Cellix flow-based adhesion assays over fibronectin-treated, TNF-alfa-stimulated HUVEC layer on chip.

BMB has synthesised a range of CXCL8 chemokine peptide mutants during her secondment to better understand the role of CXCL8 glycosaminoglycan-binding region, and the chemokine nitration on chemokine-GAG interaction, through chemotaxis assays and flow-based adhesion assays. Additionally, monoclonal antibody which can detect nitrated chemokine has been generated and tested, and is currently being further validated. This antibody will be used to detect the presence of nitrated CXCL8 in renal Transplant patient serum and urine samples. Better understanding of these effects could offer therapeutic opportunities to protect from neutrophil- derived damage during IRI.