title: Use of bioluminescence imaging to track neutrophil migration and its inhibition in experimental colitis.
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 β-actin-luciferase mice were isolated 12h 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 16-22h post-transfer. Anti-KC antibody or an isotype control were administered at 20 µg/mouse 1h before transfer, followed by whole-body and organ imaging 4h 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 4h 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.
authors: Alimentary Pharmabiotic Centre, University College Cork, National University of Ireland, Cork, Ireland.
authors: C T
authors: CT
authors: Murphy
authors: G
authors: G
authors: Moloney
authors: L J
authors: LJ
authors: Hall
authors: A
authors: A
authors: Quinlan
authors: E
authors: E
authors: Faivre
authors: P
authors: P
authors: Casey
authors: F
authors: F
authors: Shanahan
authors: S
authors: S
authors: Melgar
authors: K
authors: K
authors: Nally
mesh_terms: D000199:Actins / Q000235:genetics / Q000378:metabolism; D000818:Animals; D002465:Cell Movement; D002634:Chemotaxis, Leukocyte; D003092:Colitis / Q000139:chemically induced / Q000235:genetics / Q000378:metabolism*; D016264:Dextran Sulfate; D004195:Disease Models, Animal; D005260:Female; D005434:Flow Cytometry; D007700:Kinetics; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D049449:Luminescence; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008822:Mice, Transgenic; D009504:Neutrophils / Q000166:cytology* / Q000378:metabolism; D023063:Receptors, Interleukin-8B / Q000378:metabolism
chemical_list: D000199:Actins; D023063:Receptors, Interleukin-8B; D016264:Dextran Sulfate; D008156:Luciferases