Carbohydrates from Pseudomonas aeruginosa biofilms interact with immune C-type lectins and interfere with their receptor function

Singh, Sonali; Almuhanna, Yasir; Alshahrani, Mohammad Y.; Lowman, Douglas W.; Rice, Peter J.; Gell, Chris; Ma, Zuchao; Graves, Bridget; Jackson, Darryl; Lee, Kelly; Juarez, Rucha; Koranteng, Janice; Muntaka, Sirina; Mitchell, Daniel A.; da Silva, Ana C.; Hussain, Farah; Yilmaz, Gokhan; Mastrotto, Francesca; Irie, Yasuhiko; Williams, Paul; Williams, David L.; Cámara, Miguel; Martinez-Pomares, Luisa

Abstract

Bacterial biofilms represent a challenge to the healthcare system because of their resilience against antimicrobials and immune attack. Biofilms consist of bacterial aggregates embedded in an extracellular polymeric substance (EPS) composed of polysaccharides, nucleic acids and proteins. We hypothesised that carbohydrates could contribute to immune recognition of Pseudomonas aeruginosa biofilms by engaging C-type lectins. Here we show binding of Dendritic Cell-Specific Intercellular adhesion molecule-3-Grabbing Non-integrin (DC-SIGN, CD209), mannose receptor (MR, CD206) and Dectin-2 to P. aeruginosa biofilms. We also demonstrate that DC-SIGN, unlike MR and Dectin-2, recognises planktonic P. aeruginosa cultures and this interaction depends on the presence of the common polysaccharide antigen. Within biofilms DC-SIGN, Dectin-2 and MR ligands appear as discrete clusters with dispersed DC-SIGN ligands also found among bacterial aggregates. DC-SIGN, MR and Dectin-2 bind to carbohydrates purified from P. aeruginosa biofilms, particularly the high molecular weight fraction (HMW; >132,000 Da), with KDs in the nM range. These HMW carbohydrates contain 74.9–80.9% mannose, display α-mannan segments, interfere with the endocytic activity of cell-associated DC-SIGN and MR and inhibit Dectin-2-mediated cellular activation. In addition, biofilm carbohydrates reduce the association of the DC-SIGN ligand Lewisx, but not fucose, to human monocyte-derived dendritic cells (moDCs), and alter moDC morphology without affecting early cytokine production in response to lipopolysaccharide or P. aeruginosa cultures. This work identifies the presence of ligands for three important C-type lectins within P. aeruginosa biofilm structures and purified biofilm carbohydrates and highlights the potential for these receptors to impact immunity to P. aeruginosa infection.

Citation

Singh, S., Almuhanna, Y., Alshahrani, M. Y., Lowman, D. W., Rice, P. J., Gell, C., Ma, Z., Graves, B., Jackson, D., Lee, K., Juarez, R., Koranteng, J., Muntaka, S., Mitchell, D. A., da Silva, A. C., Hussain, F., Yilmaz, G., Mastrotto, F., Irie, Y., Williams, P., …Martinez-Pomares, L. (2021). Carbohydrates from Pseudomonas aeruginosa biofilms interact with immune C-type lectins and interfere with their receptor function. npj Biofilms and Microbiomes, 7(1), Article 87. https://doi.org/10.1038/s41522-021-00257-w

• Journal Article Type: Article
• Acceptance Date: Nov 3, 2021
• Online Publication Date: Dec 8, 2021
• Publication Date: Dec 8, 2021
• Deposit Date: Nov 10, 2021
• Publicly Available Date: Dec 8, 2021
• Journal: npj Biofilms and Microbiomes
• Electronic ISSN: 2055-5008
• Publisher: Nature Publishing Group
• Peer Reviewed: Peer Reviewed
• Volume: 7
• Issue: 1
• Article Number: 87
• DOI: https://doi.org/10.1038/s41522-021-00257-w
• Keywords: Applied Microbiology and Biotechnology; Microbiology; Biotechnology
• Public URL: https://nottingham-repository.worktribe.com/output/6677744
• Publisher URL: https://www.nature.com/articles/s41522-021-00257-w
• Additional Information: Received: 11 March 2021; Accepted: 3 November 2021; First Online: 8 December 2021; : The authors declare no competing interests.