A combined Bacteria Microarray and Quartz Crystal Microbalance approach for exploring lectin recognition of nontypeable Haemophilus influenzae

Autor: Kalograiaki, Ioanna, Euba, Begoña, Proverbio, Davide, Campanero-Rhodes, María Asunción, Aastrup, Teodor, Garmendia, Juncal, Solís, Dolores
Přispěvatelé: European Commission, Instituto de Salud Carlos III, Centro de Investigación Biomédica en Red Enfermedades Respiratorias (España), Ministerio de Economía y Competitividad (España)
Rok vydání: 2016
Zdroj: Digital.CSIC. Repositorio Institucional del CSIC
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ISSN: 2011-2890
Popis: Trabajo presentado en el III Biennal Meeting of the Chemical Biology Meeting / XII Carbohydrate Symposium, celebrado en Madrid (España), del 14 al 16 de marzo de 2016
Bacterial glycosignatures are intimately associated with virulence and regularly used for strain typification. Carbohydrate structures anchored on the bacterial surface are targeted by endogenous lectins for triggering defence responses. Elucidation of the recognition traditionally requires laborious protocols for the purification of both pathogen and host target molecules. Recently, we employed bacteria microarrays to study glycan-mediated interactions taking place at the bacterial surface, using Klebsiella pneumoniae as model[1]. In contrast to Klebsiella species, other Gram-negative bacteria are nonencapsulated or present lipooligosaccharides (LOS) instead of complex lipopolysaccharides. This is the case for nontypeable Haemophilus influenzae (NTHi), an important opportunistic pathogen responsible for respiratory and even systematic infections. In this work, we combined NTHi microarrays with the quartz crystal microbalance (QCM) technology for detection of carbohydrate epitopes and analysis of their recognition by lectins. A panel of mutant strains of the clinical isolate NTHi375 presenting a sequentially truncated LOS was examined, along with clinical isolates of different pathological origin. Reference lectins with well-defined binding specificities were used as pattern-reading tool, competition assays with lectin-specific haptens confirming carbohydrate-dependent recognition. Furthermore, the microarray-based approach was shown to be efficient for detecting strain-selective binding of three lectins of the innate immune system, namely surfactant protein D, human galectin-8 and Siglec-14. Selected bacteria-lectin pairs were analysed by QCM using novel bacteria chips, granting additional information on binding kinetics and affinity. The parameters obtained were strain- and lectin-specific, providing further insights into the interactions occurring at the bacterial surface and aiding in the establishment of functional correlations. Acknowledgements: This work was supported by the Marie Curie Initial Training Networks DYNANO (PITN-GA-2011-289033) and GLYCOPHARM (PITN-GA-2012-317297), the CIBER of Respiratory Diseases (CIBERES), an initiative from the Spanish Institute of Health Carlos III (ISCIII), and the Spanish Ministry of Economy and Competitiveness (grants BFU2012-36825 and SAF2012-31166). I.K. was funded by a Marie Curie contract from the European Commission. References [1] Campanero-Rhodes, M. A.; Llobet, E.; Bengoechea, J. A.; Solis, D. RSC Adv. 2015, 5 (10), 7173.
This work was supported by the Marie Curie Initial Training Networks DYNANO (PITN-GA-2011-289033) and GLYCOPHARM (PITN-GA-2012-317297), the CIBER of Respiratory Diseases (CIBERES), an initiative from the Spanish Institute of Health Carlos III (ISCIII), and the Spanish Ministry of Economy and Competitiveness (grants BFU2012-36825 and SAF2012-31166). I.K. was funded by a Marie Curie contract from the European Commission
Databáze: OpenAIRE
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