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

Wang G, Cochet F, Facchini FA, Zaffaroni L, Serba C, Pascal S, Andraud C, Sala A, Di Lorenzo F, Maury O, Huser T, et al. (2019)
BIOCONJUGATE CHEMISTRY 30(6): 1649-1657.

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Zeitschriftenaufsatz | Veröffentlicht | Englisch
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Abstract / Bemerkung
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.
Erscheinungsjahr
Zeitschriftentitel
BIOCONJUGATE CHEMISTRY
Band
30
Ausgabe
6
Seite(n)
1649-1657
ISSN
PUB-ID

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Wang G, Cochet F, Facchini FA, et al. Synthesis of the New Cyanine-Labeled Bacterial Lipooligosaccharides for Intracellular Imaging and in Vitro Microscopy Studies. BIOCONJUGATE CHEMISTRY. 2019;30(6):1649-1657.
Wang, G., Cochet, F., Facchini, F. A., Zaffaroni, L., Serba, C., Pascal, S., Andraud, C., et al. (2019). Synthesis of the New Cyanine-Labeled Bacterial Lipooligosaccharides for Intracellular Imaging and in Vitro Microscopy Studies. BIOCONJUGATE CHEMISTRY, 30(6), 1649-1657. doi:10.1021/acs.bioconjchem.9b00044
Wang, G., Cochet, F., Facchini, F. A., Zaffaroni, L., Serba, C., Pascal, S., Andraud, C., Sala, A., Di Lorenzo, F., Maury, O., et al. (2019). Synthesis of the New Cyanine-Labeled Bacterial Lipooligosaccharides for Intracellular Imaging and in Vitro Microscopy Studies. BIOCONJUGATE CHEMISTRY 30, 1649-1657.
Wang, G., et al., 2019. Synthesis of the New Cyanine-Labeled Bacterial Lipooligosaccharides for Intracellular Imaging and in Vitro Microscopy Studies. BIOCONJUGATE CHEMISTRY, 30(6), p 1649-1657.
G. Wang, et al., “Synthesis of the New Cyanine-Labeled Bacterial Lipooligosaccharides for Intracellular Imaging and in Vitro Microscopy Studies”, BIOCONJUGATE CHEMISTRY, vol. 30, 2019, pp. 1649-1657.
Wang, G., Cochet, F., Facchini, F.A., Zaffaroni, L., Serba, C., Pascal, S., Andraud, C., Sala, A., Di Lorenzo, F., Maury, O., Huser, T., Peri, F.: Synthesis of the New Cyanine-Labeled Bacterial Lipooligosaccharides for Intracellular Imaging and in Vitro Microscopy Studies. BIOCONJUGATE CHEMISTRY. 30, 1649-1657 (2019).
Wang, Gabriel, Cochet, Florent, Facchini, Fabio Alessandro, Zaffaroni, Lenny, Serba, Christelle, Pascal, Simon, Andraud, Chantal, Sala, Andrea, Di Lorenzo, Flaviana, Maury, Olivier, Huser, Thomas, and Peri, Francesco. “Synthesis of the New Cyanine-Labeled Bacterial Lipooligosaccharides for Intracellular Imaging and in Vitro Microscopy Studies”. BIOCONJUGATE CHEMISTRY 30.6 (2019): 1649-1657.

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