Deciphering the protein interaction in adhesion of Francisella tularensis subsp. holarctica to the endothelial cells

Bencúrová, Elena; Pulzová, Lucia; Gyuranecz, Miklós; Mlynárčik, Patrik; Mucha, Rastislav; Vlachakis, Dimitriοs; Κοσσίδα, Σοφία; Flachbartova, Zuzana; Bhide, Mangesh

doi:10.1016/j.micpath.2015.03.007

Cited by 9 publications

(6 citation statements)

References 39 publications

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“…Of note, control experiment without recombinant proteins (negative control, also used for background subtraction) confirmed the specificity of the assay. Two proteins, PilW 16 of Francisella known to not interact with BMECs and OspA 17 of neuroinvasive Borrelia known to adhere on BMEC were also included in assay. No interaction between PilW and BMEC lysate was observed (mean RFU 133.3, std.…”

Section: Resultsmentioning

confidence: 99%

“…PilW protein of Francisella (known to not interact with BMECs) and OspA of neuroinvasive Borrelia (known to interact with BMECs) were also included in the experiment as known negative and positive controls. Both proteins were produced in our laboratory previously to assess their binding affinity to BMECs 16 , 17 . ELISA was performed three times and in every experiment each recombinant protein was in triplicate.…”

Section: Methodsmentioning

confidence: 99%

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Proteomic and bioinformatic pipeline to screen the ligands of S. pneumoniae interacting with human brain microvascular endothelial cells

Jiménez-Munguía

Pulzová

Káňová

et al. 2018

Sci Rep

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The mechanisms by which Streptococcus pneumoniae penetrates the blood-brain barrier (BBB), reach the CNS and causes meningitis are not fully understood. Adhesion of bacterial cells on the brain microvascular endothelial cells (BMECs), mediated through protein-protein interactions, is one of the crucial steps in translocation of bacteria across BBB. In this work, we proposed a systematic workflow for identification of cell wall associated ligands of pneumococcus that might adhere to the human BMECs. The proteome of S. pneumoniae was biotinylated and incubated with BMECs. Interacting proteins were recovered by affinity purification and identified by data independent acquisition (DIA). A total of 44 proteins were identified from which 22 were found to be surface-exposed. Based on the subcellular location, ontology, protein interactive analysis and literature review, five ligands (adhesion lipoprotein, endo-β-N-acetylglucosaminidase, PhtA and two hypothetical proteins, Spr0777 and Spr1730) were selected to validate experimentally (ELISA and immunocytochemistry) the ligand-BMECs interaction. In this study, we proposed a high-throughput approach to generate a dataset of plausible bacterial ligands followed by systematic bioinformatics pipeline to categorize the protein candidates for experimental validation. The approach proposed here could contribute in the fast and reliable screening of ligands that interact with host cells.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Proteomic and bioinformatic pipeline to screen the ligands of S. pneumoniae interacting with human brain microvascular endothelial cells

Jiménez-Munguía

Pulzová

Káňová

et al. 2018

Sci Rep

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“…Pathological changes were studied by light microscopy on four-micrometer thick sections of formalin-fixed, paraffin-embedded tissue samples stained with haematoxylin and eosin. Immunohistochemical examinations were performed as described before [ 22 ]. In brief, sections were de-paraffinized and incubated in 3% H 2 O 2 solution for 10 min and in 2% solution of skimmed milk powder for 20 min.…”

Section: Methodsmentioning

confidence: 99%

Comparison of virulence of Francisella tularensis ssp. holarctica genotypes B.12 and B.FTNF002-00

et al. 2016

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BackgroundTwo main genetic groups (B.12 and B.FTNF002-00) of Francisella tularensis ssp. holarctica are endemic in Europe. The B.FTNF002-00 group proved to be dominant in Western European countries, while strains of the B.12 group were isolated mainly in Northern, Central and Eastern Europe. The clinical course of tularemia in the European brown hare (Lepus europaeus) also shows distinct patterns according to the geographical area. Acute course of the disease is observed in hares in Western European countries, while signs of sub-acute or chronic infection are more frequently detected in the eastern part of the continent. The aim of the present study was to examine whether there is any difference in the virulence of the strains belonging to the B.FTNF002-00 and B.12 genetic clades.ResultsExperimental infection of Fischer 344 rats was performed by intra-peritoneal injection of three dilutions of a Hungarian (B.12 genotype) and an Italian (B.FTNF002-00 genotype) F. tularensis ssp. holarctica strain. Moderate difference was observed in the virulence of the two genotypes. Significant differences were observed in total weight loss values and scores of clinical signs between the two genotypes with more rats succumbing to tularemia in groups infected with the B.FTNF002-00 genotype.ConclusionsResults of the experimental infection are consistent with previous clinical observations and pathological studies suggesting that F. tularensis ssp. holarctica genotype B.FTNF002-00 has higher pathogenic potential than the B.12 genotype.Electronic supplementary materialThe online version of this article (doi:10.1186/s12917-017-0968-9) contains supplementary material, which is available to authorized users.

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“…At some stage of dissemination into various organs, Francisella must overcome the endothelial barrier in the microvasculature by one of three well-known mechanisms: transcellular, paracellular, or the so-called Trojan horse mechanism (i.e., crossing the barrier using infected phagocytes). To achieve this, Francisella readily adheres to the endothelial cell surface and uses PilE4 (type IV pili subunit) to interact with ICAM-1 molecule, adhere to the endothelial surface, and cross the endothelial barrier in vivo as well as in vitro (Bencurova et al, 2015 ). Endothelial as well as epithelial cells and hepatocytes produce a variety of cytokines at various levels after interaction with bacteria.…”

Section: Signaling Windows Concept—spatiotemporal Network Of Cellularmentioning

confidence: 99%

Innate Immune Recognition: Implications for the Interaction of Francisella tularensis with the Host Immune System

Kročová

Macela

Kubelková

2017

Front. Cell. Infect. Microbiol.

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The intracellular bacterial pathogen Francisella tularensis causes serious infectious disease in humans and animals. Moreover, F. tularensis, a highly infectious pathogen, poses a major concern for the public as a bacterium classified under Category A of bioterrorism agents. Unfortunately, research has so far failed to develop effective vaccines, due in part to the fact that the pathogenesis of intracellular bacteria is not fully understood and in part to gaps in our understanding of innate immune recognition processes leading to the induction of adaptive immune response. Recent evidence supports the concept that immune response to external stimuli in the form of bacteria is guided by the primary interaction of the bacterium with the host cell. Based on data from different Francisella models, we present here the basic paradigms of the emerging innate immune recognition concept. According to this concept, the type of cell and its receptor(s) that initially interact with the target constitute the first signaling window; the signals produced in the course of primary interaction of the target with a reacting cell act in a paracrine manner; and the innate immune recognition process as a whole consists in a series of signaling windows modulating adaptive immune response. Finally, the host, in the strict sense, is the interacting cell.

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Deciphering the protein interaction in adhesion of Francisella tularensis subsp. holarctica to the endothelial cells

Cited by 9 publications

References 39 publications

Proteomic and bioinformatic pipeline to screen the ligands of S. pneumoniae interacting with human brain microvascular endothelial cells

Proteomic and bioinformatic pipeline to screen the ligands of S. pneumoniae interacting with human brain microvascular endothelial cells

Comparison of virulence of Francisella tularensis ssp. holarctica genotypes B.12 and B.FTNF002-00

Innate Immune Recognition: Implications for the Interaction of Francisella tularensis with the Host Immune System

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