A 63 kDa VSP9B10A-like protein expressed in a C-8 Giardia duodenalis Mexican clone

Bermúdez‐Cruz, Rosa María; Ortega‐Pierres, Guadalupe; Ceja, Víctor; Coral‐Vázquez, Ramón Mauricio; Fonseca, Rocı́o; Cervantes, Lourdes; Sanchez, Alejandra Hernandez; Depardón, Francisco; Newport, George; Montañéz, Cecilia

doi:10.1016/j.arcmed.2003.12.005

Cited by 7 publications

(4 citation statements)

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“…These conditions allow a maximum (~70%) of parasite adhesion (McCabe et al 1991). Trophozoite adhesion was quantified by thymidine-labelling of parasites as described previously (Bermúdez-Cruz et al 2004). To determine the effect of cysteine protease inhibitors on parasite adhesion, 2 × 10 6 trophozoites were pre-incubated with 1 µM E64 or 50 µM TPCK for 30 min, i.e., at concentrations below the LC 20 values for E64 and TPCK (1.05 µM and 234.94 µM respectively).…”

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Giardia duodenalis: analysis of secreted proteases upon trophozoite-epithelial cell interaction in vitro

Rodríguez-Fuentes

Cedillo‐Rivera

Fonseca-Liñán

et al. 2006

Mem. Inst. Oswaldo Cruz

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Protease secretion by Giardia duodenalis trophozoites upon interaction with epithelial cells and its association with the parasite adhesion was studied in co-cultures of parasites with IEC6 epithelial cell monolayers in the presence or absence of protease inhibitors. Proteolytic activity in supernatants from trophozoites was enhanced when they were co-cultured with IEC6 cells. This activity was strongly inhibited by pre-incubation of live trophozoites with E-64 and TPCK and a concomitant inhibition of parasite adhesion to IEC6 cells was observed. These data suggest that trophozoites secrete cysteine-type proteases that play a role in the adhesion of G. duodenalis to epithelial cells.Key words: Giardia duodenalis -protease secretion -adhesion Giardiasis caused by the flagellated, intestinal protozoan parasite Giardia duodenalis (syn. G. lamblia, G. intestinalis) manifests as acute and chronic diarrhoea in communities of both developing and developed countries. It is also a leading cause of defined waterborne diarrhoea worldwide (Adam 1991). Infection results from ingestion of cysts, most commonly from faecal contamination of water and food whereas community-wide outbreaks result from contaminated water supplies. After excystation in the upper small intestine, adhesion of trophozoites to epithelial intestinal cells represents the first step in the pathogenesis of the disease. The use of IEC6, an epithelial cell line derived from rat small intestine, is a well validated model in which the inhibitory effect of both actin inhibitors and chelating agents on the attachment of G. duodenalis trophozoites has been previously demonstrated (McCabe et al. 1991). It is also well recognized that the adhesion process is a multi-factorial event that involves a suction force of the adhesive disk (Knaippe 1990), a mechanical process related to contractile proteins of the adhesive disk and the ventrolateral flange (Chávez et al. 1995) and surface molecules that allow the binding of trophozoites to receptors to host epithelial cells (Pegado & Souza 1994). It has been postulated that proteases may be involved in this process. Several studies have demonstrated protease activity in total extracts of Giardia trophozoites or in parasite vacuoles (Feely & Dyer 1987, Hare et al. 1989) including the Portland 1 (P1) strain cultured in vitro, with the predominance of cysteine, serine, and aspartic protease activities (Williams & Coombs 1995). It has also been shown that cysteine proteases of P1 strain degraded protein substrates such as gelatin, collagen, albumin, and azocasein while serine proteases were mainly active against haemoglobin (Coradi & Guimarães 2006). In addition, the presence of cysteine protease activities in excretory/secretory (E/S) products of P1 strain has also been reported (Jiménez et al. 2000). These E/S products are associated with mucosal injuries observed in murine giardiasis (Jiménez et al. 2004). However the possible relation between protease secretion and the adhesion of G. duodenalis to epithelial cells has not b...

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confidence: 99%

Giardia duodenalis: analysis of secreted proteases upon trophozoite-epithelial cell interaction in vitro

Rodríguez-Fuentes

Cedillo‐Rivera

Fonseca-Liñán

et al. 2006

Mem. Inst. Oswaldo Cruz

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“…These include physicochemical procedures that affect homeostatic processes, namely tonic shock that deregulate cell volume recovery (Hansen and Fletcher, 2008) and mucosal barriers such as poly-anionic, heavily glycosylated proteins (mucins) that by electrostatic repulsion may impede the contact between trophozoite and substrate (Roskens and Erlandsen, 2002). rabbit antiserum, monoclonal antibodies, and secretory IgA from human milk; or to specifi c surface molecules such as a 200-kDa protein, the variable surface protein VSP9B10A-like and the cytoskeletal, ventral disk-localized protein δ-giardin (Bermúdez-Cruz et al, 2004;Hernández-Sánchez et al, 2008;. Other blocking agents are antibodies directed to cell surface components of trophozoites (e.g.…”

Section: Molecular Factors Involved In Giardia Adhesion To Host Cellsmentioning

confidence: 99%

“…Live parasites, soluble extracts (Chawla et al, 1975;Seow et al, 1993) Chymotrypsin Humans Live parasites, soluble extracts (Chawla et al, 1975;Carroccio et al, 1997) 2001), unknown Giardia cysteine proteases ( Rodriguez-Fuentes et al, 2006), and VSPs (Bermudez-Cruz et al, 2004).…”

Section: Gl50803_103676mentioning

confidence: 99%

“…Indeed, recent observations indi- (Rodriguez-Fuentes et al, 2006) VSPs (Bermudez-Cruz et al, 2004) Surface lectins δ-giardian Bermudez-Cruz et al, 2004;Rodriguez-Fuentes et al, 2006; 2. Indeed, recent observations indi- (Rodriguez-Fuentes et al, 2006) VSPs (Bermudez-Cruz et al, 2004) Surface lectins δ-giardian Bermudez-Cruz et al, 2004;Rodriguez-Fuentes et al, 2006; 2.…”

Section: Gl50803_103676mentioning

confidence: 99%

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Giardia

2011

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This work is subject to copyright. All rights are reserved, whether the whole or part of the material is concerned, specifi cally those of translation, reprinting, re-use of illustrations, broadcasting, reproduction by photocopying machines or similar means, and storage in data banks.Product Liability: The publisher can give no guarantee for all the information contained in this book. This does also refer to information about drug dosage and application thereof. In every individual case the respective user must check its accuracy by consulting other pharmaceutical literature.The use of registered names, trademarks, etc. in this publication does not imply, even in the absence of a specifi c statement, that such names are exempt from the relevant protective laws and regulations and therefore free for general use.

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