Reactive Oxygen Species Imaging in a Mouse Model of Inflammatory Bowel Disease

PurposeReactive oxygen species (ROS) are important contributors to inflammatory bowel disease (IBD); however, there are insufficient tools for their in vivo evaluation.ProceduresTo determine if a chemiluminescent ROS reporter, coelenterazine, would be a useful tool for the detection of immune cell activation, the macrophage cell line (RAW 264.7) was treated with phorbol myristate acetate (PMA). Additionally, coelenterazine was used to monitor the changes in ROS production over time in a mouse model of IBD.ResultsIn vitro, coelenterazine enabled the dynamic monitoring of the RAW 264.7 cell oxidative burst. In vivo, there were early, preclinical, changes in the localization and magnitude of coelenterazine chemiluminescent foci.ConclusionsCoelenterazine offers a high-throughput method for assessing immune cell activation in culture and provides a means for the in vivo detection and localization of ROS during IBD disease progression.

[1]  D. Piwnica-Worms,et al.  Spying on cancer: molecular imaging in vivo with genetically encoded reporters. , 2005, Cancer cell.

[2]  A. Keshavarzian,et al.  Increased production of luminol enhanced chemiluminescence by the inflamed colonic mucosa in patients with ulcerative colitis. , 1993, Gut.

[3]  K. Yasunari,et al.  Carotid artery intima-media thickness and reactive oxygen species formation by monocytes in hypertensive patients , 2006, Journal of Human Hypertension.

[4]  D. Rampton,et al.  Chemiluminescence assay of mucosal reactive oxygen metabolites in inflammatory bowel disease. , 1992, Gastroenterology.

[5]  J. Viney,et al.  Methods of Inducing Inflammatory Bowel Disease in Mice , 2009, Current protocols in pharmacology.

[6]  F. Fang Antimicrobial reactive oxygen and nitrogen species: concepts and controversies , 2004, Nature Reviews Microbiology.

[7]  C. Plieth,et al.  A coelenterazine-based luminescence assay to quantify high-molecular-weight superoxide anion scavenger activities , 2010, Nature Protocols.

[8]  Christopher H. Contag,et al.  In vivo imaging using bioluminescence: a tool for probing graft-versus-host disease , 2006, Nature Reviews Immunology.

[9]  M. Lucas,et al.  Coelenterazine is a superoxide anion-sensitive chemiluminescent probe: its usefulness in the assay of respiratory burst in neutrophils. , 1992, Analytical biochemistry.

[10]  Z. Cohn,et al.  Increased superoxide anion production by immunologically activated and chemically elicited macrophages , 1978, The Journal of experimental medicine.

[11]  A. Keshavarzian,et al.  Excessive production of reactive oxygen metabolites by inflamed colon: analysis by chemiluminescence probe. , 1992, Gastroenterology.

[12]  A. Keshavarzian,et al.  Role of reactive oxygen metabolites in experimental colitis. , 1990, Gut.

[13]  C. Nathan,et al.  Release of reactive nitrogen intermediates and reactive oxygen intermediates from mouse peritoneal macrophages. Comparison of activating cytokines and evidence for independent production. , 1988, Journal of immunology.

[14]  S. Gambhir,et al.  Optical imaging of Renilla luciferase reporter gene expression in living mice , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[15]  Christopher H Contag,et al.  Molecular imaging using labeled donor tissues reveals patterns of engraftment, rejection, and survival in transplantation. , 2005, Transplantation.

[16]  R. Löfberg,et al.  Subclinical time span of inflammatory bowel disease in patients with primary sclerosing cholangitis , 1995, Diseases of the colon and rectum.

[17]  A. Keshavarzian,et al.  Mitomycin C-induced colitis in rats: a new animal model of acute colonic inflammation implicating reactive oxygen species. , 1992, The Journal of laboratory and clinical medicine.

[18]  M. Azadniv,et al.  Imaging CD8+ T cell dynamics in vivo using a transgenic luciferase reporter. , 2007, International immunology.

[19]  M. Dubuisson,et al.  The origins of marine bioluminescence: turning oxygen defence mechanisms into deep-sea communication tools. , 1998, The Journal of experimental biology.

[20]  C. Contag,et al.  Multimodality Imaging of Cancer Superoxide Anion Using the Small Molecule Coelenterazine , 2016, Molecular Imaging and Biology.

[21]  W. Phillips,et al.  Activation of the macrophage respiratory burst by phorbol myristate acetate: evidence for both tyrosine-kinase-dependent and -independent pathways. , 1994, Biochimica et biophysica acta.

[22]  L. Mcphail,et al.  Phorbol myristate acetate induces neutrophil NADPH‐oxidase activity by two separate signal transduction pathways: dependent or independent of phosphatidylinositol 3‐kinase , 2000, Journal of leukocyte biology.

[23]  A. Szalay,et al.  Imaging of light emission from the expression of luciferases in living cells and organisms: a review. , 2002, Luminescence : the journal of biological and chemical luminescence.

[24]  Christopher H Contag,et al.  Characterization of coelenterazine analogs for measurements of Renilla luciferase activity in live cells and living animals. , 2004, Molecular imaging.

[25]  C. Contag,et al.  Chemiluminescence Imaging of Superoxide Anion Detects Beta-Cell Function and Mass , 2016, PloS one.

[26]  O. Shimomura,et al.  Coelenterazine analogs as chemiluminescent probe for superoxide anion. , 1997, Analytical biochemistry.

[27]  Irving L. Weissman,et al.  Shifting foci of hematopoiesis during reconstitution from single stem cells , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[28]  C. Nathan,et al.  Reactive oxygen and nitrogen intermediates in the relationship between mammalian hosts and microbial pathogens. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[29]  S. Reddy,et al.  Reactive oxygen species in inflammation and tissue injury. , 2014, Antioxidants & redox signaling.