Participation of heparin binding proteins from the surface of Leishmania (Viannia) braziliensis promastigotes in the adhesion of parasites to Lutzomyia longipalpis cells (Lulo) in vitro

Background: Leishmania (V.) braziliensis is a causative agent of cutaneous leishmaniasis in Brazil. During the parasite life cycle, the promastigotes adhere to the gut of sandflies, to avoid being eliminated with the dejection. The Lulo cell line, derived from Lutzomyia longipalpis (Diptera: Psychod...

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Fecha de publicación:
2012
Institución:
Universidad del Rosario
Repositorio:
Repositorio EdocUR - U. Rosario
Idioma:
eng
OAI Identifier:
oai:repository.urosario.edu.co:10336/21868
Acceso en línea:
https://doi.org/10.1186/1756-3305-5-142
https://repository.urosario.edu.co/handle/10336/21868
Palabra clave:
Enfermedades
L. (V.) braziliensis
Promastigotes
Lulo cells
Glycosaminoglycans
Surface plasmon resonance
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dc.title.spa.fl_str_mv Participation of heparin binding proteins from the surface of Leishmania (Viannia) braziliensis promastigotes in the adhesion of parasites to Lutzomyia longipalpis cells (Lulo) in vitro
title Participation of heparin binding proteins from the surface of Leishmania (Viannia) braziliensis promastigotes in the adhesion of parasites to Lutzomyia longipalpis cells (Lulo) in vitro
spellingShingle Participation of heparin binding proteins from the surface of Leishmania (Viannia) braziliensis promastigotes in the adhesion of parasites to Lutzomyia longipalpis cells (Lulo) in vitro
Enfermedades
L. (V.) braziliensis
Promastigotes
Lulo cells
Glycosaminoglycans
Surface plasmon resonance
title_short Participation of heparin binding proteins from the surface of Leishmania (Viannia) braziliensis promastigotes in the adhesion of parasites to Lutzomyia longipalpis cells (Lulo) in vitro
title_full Participation of heparin binding proteins from the surface of Leishmania (Viannia) braziliensis promastigotes in the adhesion of parasites to Lutzomyia longipalpis cells (Lulo) in vitro
title_fullStr Participation of heparin binding proteins from the surface of Leishmania (Viannia) braziliensis promastigotes in the adhesion of parasites to Lutzomyia longipalpis cells (Lulo) in vitro
title_full_unstemmed Participation of heparin binding proteins from the surface of Leishmania (Viannia) braziliensis promastigotes in the adhesion of parasites to Lutzomyia longipalpis cells (Lulo) in vitro
title_sort Participation of heparin binding proteins from the surface of Leishmania (Viannia) braziliensis promastigotes in the adhesion of parasites to Lutzomyia longipalpis cells (Lulo) in vitro
dc.subject.ddc.spa.fl_str_mv Enfermedades
topic Enfermedades
L. (V.) braziliensis
Promastigotes
Lulo cells
Glycosaminoglycans
Surface plasmon resonance
dc.subject.keyword.spa.fl_str_mv L. (V.) braziliensis
Promastigotes
Lulo cells
Glycosaminoglycans
Surface plasmon resonance
description Background: Leishmania (V.) braziliensis is a causative agent of cutaneous leishmaniasis in Brazil. During the parasite life cycle, the promastigotes adhere to the gut of sandflies, to avoid being eliminated with the dejection. The Lulo cell line, derived from Lutzomyia longipalpis (Diptera: Psychodidae), is a suitable in vitro study model to understand the features of parasite adhesion. Here, we analyze the role of glycosaminoglycans (GAGs) from Lulo cells and proteins from the parasites in this event. Methods: Flagellar (F f) and membrane (Mf) fractions from promastigotes were obtained by differential centrifugation and the purity of fractions confirmed by western blot assays, using specific antibodies for cellular compartments. Heparin-binding proteins (HBP) were isolated from both fractions using a HiTrap-Heparin column. In addition, binding of promastigotes to Lulo cells or to a heparin-coated surface was assessed by inhibition assays or surface plasmon resonance (SPR) analysis. Results: The success of promastigotes subcellular fractionation led to the obtainment of Ff and Mf proteins, both of which presented two main protein bands (65.0 and 55.0kDa) with affinity to heparin. The contribution of HBPs in the adherence of promastigotes to Lulo cells was assessed through competition assays, using HS or the purified HBPs fractions. All tested samples presented a measurable inhibition rate when compared to control adhesion rate (17 ± 2.0% of culture cells with adhered parasites): 30% (for HS 20?g/ml) and 16% (for HS 10μg/ml); HBP Mf (35.2% for 10μg/ml and 25.4% for 20μg/ml) and HBP Ff (10.0% for 10μg/ml and 31.4% for 20μg/ml). Additionally, to verify the presence of sulfated GAGs in Lulo cells surface and intracellular compartment, metabolic labeling with radioactive sulfate was performed, indicating the presence of an HS and chondroitin sulfate in both cell sections. The SPR analysis performed further confirmed the presence of GAGs ligands on L. (V.) braziliensis promastigote surfaces. Conclusions: The data presented here point to evidences that HBPs present on the surface of L. (V.) braziliensis promastigotes participate in adhesion of these parasites to Lulo cells through HS participation. © 2012 de Castro Crtes et al.; licensee BioMed Central Ltd.
publishDate 2012
dc.date.created.none.fl_str_mv 2012
dc.date.issued.none.fl_str_mv 2012
dc.date.accessioned.none.fl_str_mv 2020-05-05T20:58:48Z
dc.date.available.none.fl_str_mv 2020-05-05T20:58:48Z
dc.type.eng.fl_str_mv article
dc.type.coarversion.fl_str_mv http://purl.org/coar/version/c_970fb48d4fbd8a85
dc.type.coar.fl_str_mv http://purl.org/coar/resource_type/c_6501
dc.type.spa.spa.fl_str_mv Artículo
dc.identifier.doi.none.fl_str_mv https://doi.org/10.1186/1756-3305-5-142
dc.identifier.issn.none.fl_str_mv 1756-3305
dc.identifier.uri.none.fl_str_mv https://repository.urosario.edu.co/handle/10336/21868
url https://doi.org/10.1186/1756-3305-5-142
https://repository.urosario.edu.co/handle/10336/21868
identifier_str_mv 1756-3305
dc.language.iso.spa.fl_str_mv eng
language eng
dc.relation.citationIssue.none.fl_str_mv No. 1
dc.relation.citationTitle.none.fl_str_mv Parasites and Vectors
dc.relation.citationVolume.none.fl_str_mv Vol. 5
dc.relation.ispartof.spa.fl_str_mv Parasites and Vectors, ISSN: 1756-3305 Vol. 5, No. 1 (2012)
dc.relation.uri.spa.fl_str_mv https://parasitesandvectors.biomedcentral.com/articles/10.1186/1756-3305-5-142
dc.rights.coar.fl_str_mv http://purl.org/coar/access_right/c_abf2
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institution Universidad del Rosario
dc.source.instname.none.fl_str_mv instname:Universidad del Rosario
dc.source.reponame.none.fl_str_mv reponame:Repositorio Institucional EdocUR
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Here, we analyze the role of glycosaminoglycans (GAGs) from Lulo cells and proteins from the parasites in this event. Methods: Flagellar (F f) and membrane (Mf) fractions from promastigotes were obtained by differential centrifugation and the purity of fractions confirmed by western blot assays, using specific antibodies for cellular compartments. Heparin-binding proteins (HBP) were isolated from both fractions using a HiTrap-Heparin column. In addition, binding of promastigotes to Lulo cells or to a heparin-coated surface was assessed by inhibition assays or surface plasmon resonance (SPR) analysis. Results: The success of promastigotes subcellular fractionation led to the obtainment of Ff and Mf proteins, both of which presented two main protein bands (65.0 and 55.0kDa) with affinity to heparin. The contribution of HBPs in the adherence of promastigotes to Lulo cells was assessed through competition assays, using HS or the purified HBPs fractions. All tested samples presented a measurable inhibition rate when compared to control adhesion rate (17 ± 2.0% of culture cells with adhered parasites): 30% (for HS 20?g/ml) and 16% (for HS 10μg/ml); HBP Mf (35.2% for 10μg/ml and 25.4% for 20μg/ml) and HBP Ff (10.0% for 10μg/ml and 31.4% for 20μg/ml). Additionally, to verify the presence of sulfated GAGs in Lulo cells surface and intracellular compartment, metabolic labeling with radioactive sulfate was performed, indicating the presence of an HS and chondroitin sulfate in both cell sections. The SPR analysis performed further confirmed the presence of GAGs ligands on L. (V.) braziliensis promastigote surfaces. Conclusions: The data presented here point to evidences that HBPs present on the surface of L. (V.) braziliensis promastigotes participate in adhesion of these parasites to Lulo cells through HS participation. © 2012 de Castro Crtes et al.; licensee BioMed Central Ltd.application/pdfhttps://doi.org/10.1186/1756-3305-5-1421756-3305https://repository.urosario.edu.co/handle/10336/21868engNo. 1Parasites and VectorsVol. 5Parasites and Vectors, ISSN: 1756-3305 Vol. 5, No. 1 (2012)https://parasitesandvectors.biomedcentral.com/articles/10.1186/1756-3305-5-142Abierto (Texto Completo)http://purl.org/coar/access_right/c_abf2instname:Universidad del Rosarioreponame:Repositorio Institucional EdocUREnfermedades616600L. (V.) braziliensisPromastigotesLulo cellsGlycosaminoglycansSurface plasmon resonanceParticipation of heparin binding proteins from the surface of Leishmania (Viannia) braziliensis promastigotes in the adhesion of parasites to Lutzomyia longipalpis cells (Lulo) in vitroarticleArtículohttp://purl.org/coar/version/c_970fb48d4fbd8a85http://purl.org/coar/resource_type/c_6501Castro Côrtes, Luzia Monteiro deSouza Pereira, Mirian Claudia deSouza da Silva, FranklinSantini Pereira, Bernardo AcácioOliveira Junior, Francisco Odêncio deAraújo Soares, Renata Oliveira dePeçanha Brazil, ReginaldoToma, LenyVicente, Carolina MeloniNader, Helena BoncianiMadeira, Maria de FátimaBello, Felio J.Alves, Carlos Robertode Castro Côrtes, Luzia Monteirode Souza Pereira, Mirin Claudiada Silva, Franklin SouzaSantini Pereira, Bernardo Acáciode Oliveira Junior, Frnacisco Odênciode Araújo Soares, Renata OliveiraBrazil, Reginaldo PeçanhaToma, LenyVicente, Carolina MeloniNader, Helena Boncianide Fátima Madeira, MariaBello, Felio JAlves, Carlos RobertoORIGINALParticipation_of_heparin_binding_proteins_from_the_surface_of_Leishmania.pdfapplication/pdf747224https://repository.urosario.edu.co/bitstreams/4b0a1272-6879-4634-9ead-7f9b17baeef6/download6f77e4284688232537b4e4a349e2eb8fMD51TEXTParticipation_of_heparin_binding_proteins_from_the_surface_of_Leishmania.pdf.txtParticipation_of_heparin_binding_proteins_from_the_surface_of_Leishmania.pdf.txtExtracted texttext/plain48954https://repository.urosario.edu.co/bitstreams/9f298246-bfe2-4d6b-b633-80387763f94f/download5e6444cdf598843b563635aeea6a7b3aMD52THUMBNAILParticipation_of_heparin_binding_proteins_from_the_surface_of_Leishmania.pdf.jpgParticipation_of_heparin_binding_proteins_from_the_surface_of_Leishmania.pdf.jpgGenerated Thumbnailimage/jpeg4381https://repository.urosario.edu.co/bitstreams/854208e8-17bf-4ed6-9d10-9b2f03533dcc/download69e3243a12ae31bf286a4531b61680f4MD5310336/21868oai:repository.urosario.edu.co:10336/218682020-05-13 14:49:35.288https://repository.urosario.edu.coRepositorio institucional EdocURedocur@urosario.edu.co