Use of bioluminescence imaging to track neutrophil migration and its inhibition in experimental colitis
Murphy, Carola T.; Moloney, Gerard M.; Hall, Lindsay J.; Quinlan, Aoife; Faivre, Emilie; Casey, Pat G.; Shanahan, Fergus; Melgar, Silvia; Nally, Kenneth
Date:
2010-10
Copyright:
© 2010 The Authors. Clinical and Experimental Immunology © 2010 British Society for Immunology. The definitive version is available at www.blackwell-synergy.com
Citation:
Murphy, C. T., Moloney, G., Hall, L. J., Quinlan, A., Faivre, E., Casey, P., Shanahan, F., Melgar, S. and Nally, K. (2010) 'Use of bioluminescence imaging to track neutrophil migration and its inhibition in experimental colitis', Clinical & Experimental Immunology, 162(1), pp. 188-196. doi: 10.1111/j.1365-2249.2010.04234.x
Abstract:
Inflammatory bowel disease (IBD) is associated with neutrophil infiltration into the mucosa and crypt abscesses. The chemokine interleukin (IL)-8 [murine homologues (KC) and macrophage inflammatory protein (MIP)-2] and its receptor CXCR2 are required for neutrophil recruitment; thus, blocking this engagement is a potential therapeutic strategy. In the present study, we developed a preclinical model of neutrophil migration suitable for investigating the biology of and testing new drugs that target neutrophil trafficking. Peritoneal exudate neutrophils from transgenic beta-actin-luciferase mice were isolated 12 h after intraperitoneal injection with thioglycollate, and were assessed phenotypically and functionally. Exudate cells were injected intravenously into recipients with dextran sodium sulphate (DSS)-induced colitis followed by bioluminescence imaging of whole-body and ex vivo organs at 2, 4 and 16-22 h post-transfer. Anti-KC antibody or an isotype control were administered at 20 mu g/mouse 1 h before transfer, followed by whole-body and organ imaging 4 h post-transfer. The peritoneal exudate consisted of 80% neutrophils, 39% of which were CXCR2+. In vitro migration towards KC was inhibited by anti-KC. Ex vivo bioluminescent imaging showed that neutrophil trafficking into the colon of DSS recipients was inhibited by anti-KC 4 h post-cell transfer. In conclusion, this study describes a new approach for investigating neutrophil trafficking that can be used in preclinical studies to evaluate potential inhibitors of neutrophil recruitment.
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