Extracellular protease profile of Acanthamoeba after prolonged axenic culture and after interaction with MDCK cells

Cirelli, Cecília; Mesquita, Elaine Isabela Soares; Chagas, Isabela Aurora Rodrigues; Furst, Cinthia; Possamai, Cynara Oliveira; Abrahão, Jônatas Santos; Silva, Ludmila Karen dos Santos; Grossi, Marina Felipe; Tagliati, Carlos Alberto; Costa, Adriana Oliveira

doi:10.1007/s00436-019-06562-w

Cited by 14 publications

(10 citation statements)

References 42 publications

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“…The disappearance of a 47‐kDa band also indicated the existence of cysteine and metalloproteases in the EVs cargo. These findings corroborate previous studies that showed predominance of serine proteases and the presence in lower amount of metallo‐ and cysteine proteases in Acanthamoeba , either in conditioned medium (Alsam et al, 2005; Cirelli et al, 2020; Khan, 2006; Serrano‐Luna et al, 2006; Sissons et al 2005; Ramírez‐Rico et al, 2015), or in EVs (Gonçalves et al, 2018; Retana‐Moreira et al, 2020).…”

Section: Discussionsupporting

confidence: 90%

“…In theory, variation in secretoma of distinct Acanthamoeba strains could reflect variations in their virulence. This has been suggested in studies relating higher activity of excreted proteases in pathogenic strains compared to nonpathogenic (Cirelli et al, 2020; Khan et al, 2000). However, the investigations addressing proteases activity in EVs cargo have used only a single strain (Gonçalves et al, 2018; Lin et al, 2019; Retana‐Moreira et al, 2020), reinforcing the need of expanding the number of isolates for comparison.…”

Section: Discussionmentioning

confidence: 71%

“… Note : Genotypic and physiological characterization described in (a) Duarte et al (2013), (b) Cirelli et al (2020), (c) Possamai et al (2018), and (d) Da Rocha‐Azevedo and Costa e Silva‐Filho (2007), (e) Verissimo et al (2013). …”

Section: Methodsmentioning

confidence: 99%

“…This early interaction will result in the release of proteases, which are determinant for extracellular matrix disruption and cell death by necrosis and apoptosis (Clarke et al, 2006; Neelam & Niederkorn, 2017). Acanthamoeba can predominantly produce serine proteases and less amounts of cysteine and metalloproteases with instraspecies polymorphisms (Cirelli et al, 2020; Khan, 2006; Serrano‐Luna et al, 2006; Sissons et al, 2006). Proteases can induce interleukin (IL)‐12 production via the protease‐activated receptor 1 (PAR 1 ) in murine macrophages and IL‐6 via a PAR‐independent pathway (Cano et al, 2019).…”

Section: Introductionmentioning

confidence: 99%

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Distinct immunomodulatory properties of extracellular vesicles released by different strains of Acanthamoeba

Costa

Chagas

Menezes-Neto

et al. 2021

Cell Biology International

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Free living amoeba of the genus Acanthamoeba are opportunist protozoan involved in corneal, systemic, and encephalic infections in humans. Most of the mechanisms underlying intraspecies variations and pathogenicity are still unknown. Recently, the release of extracellular vesicles (EVs) by Acanthamoeba was reported. However, comparative characterization of EVs from distinct strains is not available. The aim of this study was to evaluate EVs produced by Acanthamoeba from different genotypes, comparing their proteases profile and immunomodulatory properties. EVs from four environmental or clinical strains (genotypes T1, T2, T4, and T11) were obtained by ultracentrifugation, quantitated by nanoparticle tracking analysis and analyzed by scanning and transmission electron microscopy. Proteases profile was determined by zymography and functional properties of EVs (measure of nitrite and cytokine production) were determined after peritoneal macrophage stimulation. Despite their genotype, all strains released EVs and no differences in size and/or concentration were detected. EVs exhibited a predominant activity of serine proteases (pH 7.4 and 3.5), with higher intensity in T4 and T1 strains. EVs from the environmental, nonpathogenic T11 strain exhibited a more proinflammatory profile, inducing higher levels of Nitrite, tumor necrosis factor alpha and interleukin‐6 via TLR4/TLR2 than those strains with pathogenic traits (T4, T1, and T2). Preincubation with EVs treated with protease inhibitors or heating drastically decreased nitrite concentration production in macrophages. Those data suggest that immunomodulatory effects of EVs may reflect their pathogenic potential depending on the Acanthamoeba strains and are dependent on protease integrity.

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Section: Discussionsupporting

confidence: 90%

Section: Discussionmentioning

confidence: 71%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Distinct immunomodulatory properties of extracellular vesicles released by different strains of Acanthamoeba

Costa

Chagas

Menezes-Neto

et al. 2021

Cell Biology International

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“…This genotype is rare, but it has been associated with cases of GAE, being the second in relevance in this disease (Alsam et al ., 2005; Sissons et al ., 2006; Maciver et al ., 2013). Additionally, this strain presents high protease activity in conditioned medium, a characteristic linked to pathogenicity (Cirelli et al ., 2020).…”

Section: Discussionmentioning

confidence: 99%

Acanthamoeba spp. monoclonal antibody against a CPA2 transporter: a promising molecular tool for acanthamoebiasis diagnosis and encystment study

et al. 2020

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Free-living amoeba of the genus Acanthamoeba are ubiquitous protozoa involved in opportunistic and non-opportunistic infection in humans, such as granulomatous amoebic encephalitis and amoebic keratitis. Both infections have challenging characteristics such as the formation of the resistant cysts in infected tissues, hampering the treatment and most usual diagnosis depending on time-consuming and/or low sensitivity techniques. The use of monoclonal antibodies presents itself as an opportunity for the development of more effective alternative diagnostic methods, as well as an important and useful tool in the search for new therapeutic targets. This study investigated the possibility of using a previously produced monoclonal antibody (mAb3), as a diagnostic tool for the detection of Acanthamoeba trophozoites by direct and indirect flow cytometry and immunofluorescence. Immunoprecipitation assay and mass spectrometry allowed the isolation of the antibody's target and suggested it is a transporter part of the CPA (cation: proton antiporter) superfamily. In vitro tests indicate an important role of this target in Acanthamoeba's encystment physiology. Our results support the importance of studying the role of CPA2 transporters in the context of acanthamoebiasis, as this may be a way to identify new therapeutic candidates.

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Experimental keratitis induced in rat by Acanthamoeba from distinct morphological groups/genotypes: a histological and immunohistochemical evaluation

et al. 2023

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Extracellular protease profile of Acanthamoeba after prolonged axenic culture and after interaction with MDCK cells

Cited by 14 publications

References 42 publications

Distinct immunomodulatory properties of extracellular vesicles released by different strains of Acanthamoeba

Distinct immunomodulatory properties of extracellular vesicles released by different strains of Acanthamoeba

Acanthamoeba spp. monoclonal antibody against a CPA2 transporter: a promising molecular tool for acanthamoebiasis diagnosis and encystment study

Experimental keratitis induced in rat by Acanthamoeba from distinct morphological groups/genotypes: a histological and immunohistochemical evaluation

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