Bacterial Surface Glycans: Microarray and QCM Strategies for Glycophenotyping and Exploration of Recognition by Host Receptors

Kalograiaki, Ioanna; Campanero-Rhodes, María Asunción; Proverbio, Davide; Euba, Begoña; Garmendia, Junkal; Aastrup, Teodor; Solı́s, Dolores

doi:10.1016/bs.mie.2017.06.011

Cited by 10 publications

(18 citation statements)

References 36 publications

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“…Bacterial cells, purified LPS/LOSs, and control glycoproteins were printed on either single-pad or 16-pad nitrocellulose-coated glass slides (Maine mfg FAST-slides or Grace Biolabs ONCYTE NOVA) using a manual glass-slide arraying system (V&P Scientific) or a non-contact arrayer (Sprint, Arrayjet Ltd.), respectively, essentially as described [19,22]. Probes were printed as triplicates in a dose-response format.…”

Section: Microarray Binding Assaysmentioning

confidence: 99%

Exploration of Galectin Ligands Displayed on Gram-Negative Respiratory Bacterial Pathogens with Different Cell Surface Architectures

et al. 2021

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Galectins bind various pathogens through recognition of distinct carbohydrate structures. In this work, we examined the binding of four human galectins to the Gram-negative bacteria Klebsiella pneumoniae (Kpn) and non-typeable Haemophilus influenzae (NTHi), which display different surface glycans. In particular, Kpn cells are covered by a polysaccharide capsule and display an O-chain-containing lipopolysaccharide (LPS), whereas NTHi is not capsulated and its LPS, termed lipooligosacccharide (LOS), does not contain O-chain. Binding assays to microarray-printed bacteria revealed that galectins-3, -4, and -8, but not galectin-1, bind to Kpn and NTHi cells, and confocal microscopy attested binding to bacterial cells in suspension. The three galectins bound to array-printed Kpn LPS. Moreover, analysis of galectin binding to mutant Kpn cells evidenced that the O-chain is the docking point for galectins on wild type Kpn. Galectins-3, -4, and -8 also bound the NTHi LOS. Microarray-assisted comparison of the binding to full-length and truncated LOSs, as well as to wild type and mutant cells, supported LOS involvement in galectin binding to NTHi. However, deletion of the entire LOS oligosaccharide chain actually increased binding to NTHi cells, indicating the availability of other ligands on the bacterial surface, as similarly inferred for Kpn cells devoid of both O-chain and capsule. Altogether, the results illustrate galectins’ versatility for recognizing different bacterial structures, and point out the occurrence of so far overlooked galectin ligands on bacterial surfaces.

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Section: Microarray Binding Assaysmentioning

confidence: 99%

Exploration of Galectin Ligands Displayed on Gram-Negative Respiratory Bacterial Pathogens with Different Cell Surface Architectures

et al. 2021

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“…More recently, different lectin microarray and biosensor approaches for bacteria typing and strain discrimination have been reported 19 – 22 . Following the inverse strategy, we developed novel bacteria-based microarrays and quartz crystal microbalance (QCM) chips for the screening of bacterial glycosignatures, by testing the binding of a panel of lectins with diverse carbohydrate-binding specificities, and quantitative analysis of lectin−bacteria interactions 23 – 25 . Using this combined approach, different lectin-binding fingerprints were observed for six clinical isolates of nontypeable (non-capsulated) H. influenzae (NTHi), consistent with the above mentioned inter-strain heterogeneity of the bacterium 24 .…”

Section: Introductionmentioning

confidence: 99%

Differential recognition of Haemophilus influenzae whole bacterial cells and isolated lipooligosaccharides by galactose-specific lectins

Kalograiaki

Euba

Fernández‐Alonso

et al. 2018

Sci Rep

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Bacterial surfaces are decorated with carbohydrate structures that may serve as ligands for host receptors. Based on their ability to recognize specific sugar epitopes, plant lectins are extensively used for bacteria typing. We previously observed that the galactose-specific agglutinins from Ricinus communis (RCA) and Viscum album (VAA) exhibited differential binding to nontypeable Haemophilus influenzae (NTHi) clinical isolates, their binding being distinctly affected by truncation of the lipooligosaccharide (LOS). Here, we examined their binding to the structurally similar LOS molecules isolated from strains NTHi375 and RdKW20, using microarray binding assays, saturation transfer difference NMR, and molecular dynamics simulations. RCA bound the LOSRdKW20 glycoform displaying terminal Galβ(1,4)Glcβ, whereas VAA recognized the Galα(1,4)Galβ(1,4)Glcβ epitope in LOSNTHi375 but not in LOSRdKW20, unveiling a different presentation. Binding assays to whole bacterial cells were consistent with LOSNTHi375 serving as ligand for VAA, and also suggested recognition of the glycoprotein HMW1. Regarding RCA, comparable binding to NTHi375 and RdKW20 cells was observed. Interestingly, an increase in LOSNTHi375 abundance or expression of HMW1 in RdKW20 impaired RCA binding. Overall, the results revealed that, besides the LOS, other carbohydrate structures on the bacterial surface serve as lectin ligands, and highlighted the impact of the specific display of cell surface components on lectin binding.

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“…Immediately before printing, samples were gently centrifuged (200g for 1 min) to avoid the presence of bubbles, which would cause problems in sample aspiration, and of undesired particles that could block the microarrayer capillaries. 19 Printed slides were scanned to monitor spot location and grid Cy3 signals (excitation at 532 nm, green laser) using a GenePix Autoloader 4200AL (Axon Instruments) microarray scanner. The slides can be stored at room temperature in a dry and dark place, and remain stable for at least 1 year.…”

Section: Methodsmentioning

confidence: 99%

Development and Evaluation of a Microarray Platform for Detection of Serum Antibodies Against Streptococcus pneumoniae Capsular Polysaccharides

et al. 2020

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Streptococcus pneumoniae is responsible for severe infections, causing millions of deaths yearly.Immunoglobulin G (IgG) antibodies against the capsular polysaccharide (CPS) offer S. pneumoniae serotype-specific protection. In this work, we examined the applicability of the microarray technology to detect CPS type-specific IgGs in serum, using a collection of 22 microarray-printed S. pneumoniae CPSs. First, printing of five CPSs onto nitrocellulose-coated glass slides was tested. Successful printing was only achieved for certain CPS types and concentrations. This behavior was tentatively related with diverse viscosities of the CPS solutions. Measurement of dynamic viscosities fully supported this assumption and helped to establish suitable CPS type-and concentration-dependent printing conditions. Next, the potential of CPS microarrays for detecting recognition by anti-CPS IgGs was examined using well-defined rabbit pneumococcal antisera. In all cases, the expected antiserum-CPS binding signals were detected, prompting a proof-of-concept analysis of human serum samples.Clearly distinct serum-and CPS-specific binding patterns and intensities were observed, evidencing

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Bacterial Surface Glycans: Microarray and QCM Strategies for Glycophenotyping and Exploration of Recognition by Host Receptors

Cited by 10 publications

References 36 publications

Exploration of Galectin Ligands Displayed on Gram-Negative Respiratory Bacterial Pathogens with Different Cell Surface Architectures

Exploration of Galectin Ligands Displayed on Gram-Negative Respiratory Bacterial Pathogens with Different Cell Surface Architectures

Differential recognition of Haemophilus influenzae whole bacterial cells and isolated lipooligosaccharides by galactose-specific lectins

Development and Evaluation of a Microarray Platform for Detection of Serum Antibodies Against Streptococcus pneumoniae Capsular Polysaccharides

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