title: Dual-Color Bioluminescence Imaging for Simultaneous Monitoring of the Intestinal Persistence of Lactobacillus plantarum and Lactococcus lactis in Living Mice.
abstract: Lactic acid bacteria are found in the gastrointestinal tract of mammals and have received tremendous attention due to their health-promoting properties. We report the development of two dual-color luciferase-producing Lactobacillus (Lb.) plantarum and Lactococcus (Lc.) lactis strains for noninvasive simultaneous tracking in the mouse gastrointestinal tract. We previously described the functional expression of the red luciferase mutant (CBRluc) from Pyrophorus plagiophthalamus in Lb. plantarum NCIMB8826 and Lc. lactis MG1363 (C. Daniel, S. Poiret, V. Dennin, D. Boutillier, and B. Pot, Appl Environ Microbiol 79:1086-1094, 2013, http://dx.doi.org/10.1128/AEM.03221-12). In this study, we determined that CBRluc is a better-performing luciferase for in vivo localization of both lactic acid bacteria after oral administration than the green click beetle luciferase mutant construct developed in this study. We further established the possibility to simultaneously detect red- and green-emitting lactic acid bacteria by dual-wavelength bioluminescence imaging in combination with spectral unmixing. The difference in spectra of light emission by the red and green click beetle luciferase mutants and dual bioluminescence detection allowed in vitro and in vivo quantification of the red and green emitted signals; thus, it allowed us to monitor the dynamics and fate of the two bacterial populations simultaneously. Persistence and viability of both strains simultaneously administered to mice in different ratios was studied in vivo in anesthetized mice and ex vivo in mouse feces. The application of dual-luciferase-labeled bacteria has considerable potential to simultaneously study the interactions and potential competitions of different targeted bacteria and their hosts.
authors.affiliation: Lactic Acid Bacteria and Mucosal Immunity Team, Institut Pasteur de Lille, Center for Infection and Immunity of Lille, Lille, France Université de Lille, Lille, France CNRS, UMR 8204, Lille, France catherine.daniel@ibl.cnrs.fr.
authors.forename: Catherine
authors.initials: C
authors.lastname: Daniel
authors.affiliation: Lactic Acid Bacteria and Mucosal Immunity Team, Institut Pasteur de Lille, Center for Infection and Immunity of Lille, Lille, France Université de Lille, Lille, France CNRS, UMR 8204, Lille, France.
authors.forename: Sabine
authors.initials: S
authors.lastname: Poiret
authors.affiliation: Lactic Acid Bacteria and Mucosal Immunity Team, Institut Pasteur de Lille, Center for Infection and Immunity of Lille, Lille, France Université de Lille, Lille, France CNRS, UMR 8204, Lille, France.
authors.forename: Véronique
authors.initials: V
authors.lastname: Dennin
authors.affiliation: Lactic Acid Bacteria and Mucosal Immunity Team, Institut Pasteur de Lille, Center for Infection and Immunity of Lille, Lille, France Université de Lille, Lille, France CNRS, UMR 8204, Lille, France.
authors.forename: Denise
authors.initials: D
authors.lastname: Boutillier
authors.affiliation: Université de Lille, Lille, France CNRS, UMR 8204, Lille, France BioImaging Center Lille-Nord de France, Lille, France Cellular Microbiology and Physics of Infection, Institut Pasteur de Lille, Center for Infection and Immunity of Lille, Lille, France.
authors.forename: Delphine Armelle
authors.initials: DA
authors.lastname: Lacorre
authors.affiliation: Lactic Acid Bacteria and Mucosal Immunity Team, Institut Pasteur de Lille, Center for Infection and Immunity of Lille, Lille, France Université de Lille, Lille, France CNRS, UMR 8204, Lille, France.
authors.forename: Benoit
authors.initials: B
authors.lastname: Foligné
authors.affiliation: Lactic Acid Bacteria and Mucosal Immunity Team, Institut Pasteur de Lille, Center for Infection and Immunity of Lille, Lille, France Université de Lille, Lille, France CNRS, UMR 8204, Lille, France.
authors.forename: Bruno
authors.initials: B
authors.lastname: Pot
mesh_terms: D000818:Animals; D003116:Color; D041981:Gastrointestinal Tract / Q000382:microbiology*; D017930:Genes, Reporter; D048191:Lactiplantibacillus plantarum / Q000201:enzymology / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D013294:Lactococcus lactis / Q000201:enzymology / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D008156:Luciferases / Q000032:analysis* / Q000235:genetics; D008163:Luminescent Measurements / Q000379:methods*; D051379:Mice; D050296:Microbial Viability; D013194:Staining and Labeling
chemical_list: D008156:Luciferases