Synthesis of the New Cyanine-Labeled Bacterial Lipooligosaccharides for Intracellular Imaging and in Vitro Microscopy Studies

Autor: Andrea Sala, Chantal Andraud, Fabio A. Facchini, Francesco Peri, Flaviana Di Lorenzo, Thomas R Huser, Olivier Maury, Tung Cheng Wang, Simon Pascal, Lenny Zaffaroni, Florent Cochet, Christelle Serba
Přispěvatelé: Department of Biotechnology and Biosciences, Università degli Studi di Milano-Bicocca = University of Milano-Bicocca (UNIMIB), Institut de Science et d'ingénierie supramoléculaires (ISIS), Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Matériaux et Nanosciences Grand-Est (MNGE), Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Réseau nanophotonique et optique, Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Centre Interdisciplinaire de Nanoscience de Marseille (CINaM), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Chimie - UMR5182 (LC), École normale supérieure de Lyon (ENS de Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Department of Chemical Sciences, Complesso Universitario Monte Sant'Angelo, University of Naples Federico II = Università degli studi di Napoli Federico II, Wang, T, Cochet, F, Facchini, F, Zaffaroni, L, Serba, C, Pascal, S, Andraud, C, Sala, A, Di Lorenzo, F, Maury, O, Huser, T, Peri, F, Università degli Studi di Milano-Bicocca [Milano] (UNIMIB), Réseau nanophotonique et optique, Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Matériaux et nanosciences d'Alsace (FMNGE), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA), Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU), Centre National de la Recherche Scientifique (CNRS)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-École normale supérieure - Lyon (ENS Lyon)-Institut de Chimie du CNRS (INC), Università degli studi di Napoli Federico II
Jazyk: angličtina
Rok vydání: 2019
Předmět:
Lipopolysaccharides
Lipopolysaccharide
media_common.quotation_subject
Fluorescent Dye
Biomedical Engineering
Pharmaceutical Science
Bioengineering
02 engineering and technology
Carbocyanine
In Vitro Techniques
01 natural sciences
chemistry.chemical_compound
CHIM/06 - CHIMICA ORGANICA
Fluorescence microscope
[SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry
Molecular Biology/Biochemistry [q-bio.BM]
Internalization
ComputingMilieux_MISCELLANEOUS
media_common
Fluorescent Dyes
Pharmacology
Microscopy
Bacteria
Endocytosi
010405 organic chemistry
Chemistry
In Vitro Technique
[CHIM.ORGA]Chemical Sciences/Organic chemistry
Organic Chemistry
Colocalization
Carbocyanines
021001 nanoscience & nanotechnology
Subcellular localization
Endocytosis
0104 chemical sciences
Cell biology
Toll-Like Receptor 4
CHIM/08 - CHIMICA FARMACEUTICA
TLR4
0210 nano-technology
Bacterial outer membrane
Intracellular
Biotechnology
Zdroj: Bioconjugate Chemistry
Bioconjugate Chemistry, 2019, ⟨10.1021/acs.bioconjchem.9b00044⟩
Bioconjugate Chemistry, American Chemical Society, 2019, ⟨10.1021/acs.bioconjchem.9b00044⟩
ISSN: 1043-1802
1520-4812
Popis: Endotoxin (lipooligosaccharide, LOS, and lipopolysaccharide, LPS) is the major molecular component of Gram-negative bacteria outer membrane, and very potent pro-inflammatory substance. Visualizing and tracking the distribution of the circulating endotoxin is one of the fundamental approaches to understand the molecular aspects of infection with subsequent inflammatory and immune responses, LPS also being a key player in the molecular dialogue between microbiota and host. While fluorescently labeled LPS has previously been used to track its subcellular localization and colocalization with TLR4 receptor and downstream effectors, our knowledge on lipopolysaccharide (LOS) localization and cellular activity remains almost unexplored. In this study, LOS was labeled with a novel fluorophore, Cy7N, featuring a large Stokes-shifted emission in the deep-red spectrum resulting in lower light scattering and better imaging contrast. The LOS-Cy7N chemical identity was determined by mass spectrometry, and immunoreactivity of the conjugate was evaluated. Interestingly, its application to microscopic imaging showed a faster cell internalization compared to LPS-Alexa488, despite that it is also CD14-dependent and undergoes the same endocytic pathway as LPS toward lysosomal detoxification. Our results suggest the use of the new infrared fluorophore Cy7N for cell imaging of labeled LOS by confocal fluorescence microscopy, and propose that LOS is imported in the cells by mechanisms different from those responsible for LPS uptake.
Databáze: OpenAIRE
Externí odkaz: