title: Detection of leukotriene receptor CysLT1R in inflammatory diseases by molecular imaging with near-infrared fluorescence-based contrast agents.
abstract: As leukotriene D4 receptor CysLT1R upregulation is an early event in inflammatory processes, specific detection of CysLT1R via molecular imaging might be a promising diagnostic tool for inflammatory diseases. We coupled a specific anti-CysLT1R IgG antibody to near-infrared (NIR) hemicyanine fluorophore DY-734. The fluorophore was also coupled to unspecific rabbit-IgG antibody or corresponding Fab fragments. Expression of CysLT1R in HL-60 human promyelocytic leukemia cells in vitro could be proven by reverse transcriptase-polymerase chain reaction (PCR), real-time PCR, and flow cytometry. Detection of the probes by flow cytometry showed that CysLT1R*DY-734 probe binds distinctly stronger to HL-60 cells than IgG*DY-734. Induction of ear edema in mice was conducted to test signaling of the synthesized probes in vivo. A markedly higher fluorescence intensity was observed in the edematous region than in the healthy region by a whole-body imaging system. Semiquantitative analysis showed that CysLT1R*DY-734 and Fab-CysLT1R*DY-734 probes bind 1.9- and 1.2-fold stronger, respectively, than the unspecific probes. Biodistribution studies revealed an enrichment of full-length IgG probes in liver and spleen, whereas Fab-containing probes are mostly found in liver and kidneys. Taken together, we present an approach that might improve early diagnosis of inflammatory diseases in the long term.
authors: Institute of Diagnostic and Interventional Radiology, Department of Experimental Radiology, University Hospital Jena, Jena, Germany. Corinna.Busch@med.uni-jena.de
authors: Corinna
authors: C
authors: Busch
authors: Marta
authors: M
authors: Passon
authors: Matthias
authors: M
authors: Wenzel
authors: Ines
authors: I
authors: Socher
authors: Werner A
authors: WA
authors: Kaiser
authors: Ingrid
authors: I
authors: Hilger
mesh_terms: D000818:Animals; D003287:Contrast Media; D004487:Edema / Q000378:metabolism* / Q000473:pathology; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000737:chemistry / Q000378:metabolism; D018922:HL-60 Cells; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D007249:Inflammation / Q000378:metabolism* / Q000473:pathology; D007259:Infrared Rays; D008297:Male; D051379:Mice; D057054:Molecular Imaging / Q000379:methods*; D011817:Rabbits; D018077:Receptors, Leukotriene / Q000378:metabolism*; D014018:Tissue Distribution
chemical_list: D003287:Contrast Media; D005456:Fluorescent Dyes; D007074:Immunoglobulin G; D018077:Receptors, Leukotriene; C083169:leukotriene D4 receptor