C-Type Lectin Receptor (CLR)-Fc Fusion Proteins As Tools to Screen for Novel CLR/Bacteria Interactions: An Exemplary Study on Preselected Campylobacter jejuni Isolates

Frontiers in Immunology
Sabine MayerBernd Lepenies

Abstract

C-type lectin receptors (CLRs) are carbohydrate-binding receptors that recognize their ligands often in a Ca2+-dependent manner. Upon ligand binding, myeloid CLRs in innate immunity trigger or inhibit a variety of signaling pathways, thus initiating or modulating effector functions such as cytokine production, phagocytosis, and antigen presentation. CLRs bind to various pathogens, including viruses, fungi, parasites, and bacteria. The bacterium Campylobacter jejuni (C. jejuni) is a very frequent Gram-negative zoonotic pathogen of humans, causing severe intestinal symptoms. Interestingly, C. jejuni expresses several glycosylated surface structures, for example, the capsular polysaccharide (CPS), lipooligosaccharide (LOS), and envelope proteins. This "Methods" paper describes applications of CLR-Fc fusion proteins to screen for yet unknown CLR/bacteria interactions using C. jejuni as an example. ELISA-based detection of CLR/bacteria interactions allows a first prescreening that is further confirmed by flow cytometry-based binding analysis and visualized using confocal microscopy. By applying these methods, we identified Dectin-1 as a novel CLR recognizing two selected C. jejuni isolates with different LOS and CPS genotypes. In co...Continue Reading

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Citations

May 5, 2020·The Journal of Infectious Diseases·Theodore J KottomAndrew H Limper
Aug 1, 2020·Frontiers in Cellular and Infection Microbiology·Malgorzata E MnichNina M van Sorge
Oct 27, 2018·Frontiers in Microbiology·Nuria Castro-BravoPatricia Ruas-Madiedo
Jul 24, 2020·International Journal of Molecular Sciences·Dimitri Leonid Lindenwald, Bernd Lepenies
May 2, 2019·Frontiers in Microbiology·Mariano Prado AcostaMaria Guadalupe Martinez
Jul 13, 2019·Frontiers in Immunology·Mariano MalamudMaría de Los Ángeles Serradell
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Mar 19, 2021·Frontiers in Immunology·Mariano Prado AcostaBernd Lepenies
Apr 19, 2021·Journal of Controlled Release : Official Journal of the Controlled Release Society·Verónica DuránUlrich Kalinke
Aug 28, 2021·International Journal of Molecular Sciences·Ankur MidhaSusanne Hartmann
Aug 21, 2021·Journal of Applied Microbiology·Diego C CarneiroSuzana T Cunha Lima
Oct 11, 2020·Nature Communications·Emily RodriguesMatthew S Macauley
Dec 11, 2021·Journal of Medical Microbiology·Theodore J KottomAndrew H Limper

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Methods Mentioned

BETA
confocal microscopy
transfection
electrophoresis
glycosylation
ELISA
Assay
flow cytometry

Software Mentioned

GraphPad Prism
Mincle
FlowJo

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