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Dual-Color Bioluminescence Imaging for Simultaneous Monitoring of the Intestinal Persistence of Lactobacillus plantarum and Lactococcus lactis in Living Mice

Catherine Daniel 1, * Sabine Poiret 1 Véronique Dennin 1 Denise Boutillier 1 Delphine Armelle Lacorre 2, 3 Benoit Foligné 1 Bruno Pot 1
* Corresponding author
1 Lactic Acid Bacteria & Mucosal Immunity - CIIL
CIIL - Centre d’Infection et d’Immunité de Lille - INSERM U 1019 - UMR 9017 - UMR 8204
3 CMPI - Cellular Microbiology and Physics of Infection Group [Lille]
CIIL - Centre d’Infection et d’Immunité de Lille - INSERM U 1019 - UMR 9017 - UMR 8204
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.
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https://hal.archives-ouvertes.fr/hal-03300717
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Submitted on : Tuesday, July 27, 2021 - 11:31:25 AM
Last modification on : Tuesday, October 19, 2021 - 11:56:33 PM

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Catherine Daniel, Sabine Poiret, Véronique Dennin, Denise Boutillier, Delphine Armelle Lacorre, et al.. Dual-Color Bioluminescence Imaging for Simultaneous Monitoring of the Intestinal Persistence of Lactobacillus plantarum and Lactococcus lactis in Living Mice. Applied and Environmental Microbiology, American Society for Microbiology, 2015, 81 (16), pp.5344-5349. ⟨10.1128/aem.01042-15⟩. ⟨hal-03300717⟩

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