<i>Leishmania (Viannia) braziliensis</i>: insights on subcellular distribution and biochemical properties of heparin-binding proteins

Côrtes, Luzia Monteiro de Castro; Pereira, Mirian Claudia de Souza; Oliveira, Francisco Odencio Rodrigues de; Cortê-Real, Suzana; Silva, Franklin Souza Da; Pereira, Bernardo Acácio Santini; Madeira, Maria de Fátima; Moraes, Marcia Terezinha Baroni de; Brazil, Reginaldo Peçanha; Alves, Carlos Roberto

doi:10.1017/s0031182011001910

Cited by 21 publications

(28 citation statements)

References 51 publications

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“…Leishmania promastigotes are deposited in the skin of their hosts by infected sand flies, and several Leishmania partners are found either in the dermal-epidermal junction (collagen IV, ECM-1 [36], heparan sulfate) or in the dermis (e.g., hyaluronan and collagens I, V, and VI). Most Leishmania promastigotes assayed in this study interacted with heparin, a model compound for heparan sulfate, in agreement with previous reports (9,17). However, several Leishmania strains that bound to heparin did not bind to heparan sulfate.…”

Section: Discussionsupporting

confidence: 92%

“…Indeed, most binding studies performed so far have been performed with only one strain of Leishmania and few host molecules (5,6). Promastigotes were not fixed with paraformaldehyde before the SPRi assays, in contrast to those used for studying the interaction of L. braziliensis to Lulo cells or to a heparincoated surface by SPR (17,34).…”

Section: Discussionmentioning

confidence: 99%

“…Previous studies on the interactions established by Leishmania with some extracellular matrix components were performed with various forms of the parasites, such as amastigotes (11), promastigotes near or at the stationary phase (15,16), stationary-phase promastigotes (5) and promastigotes fixed with 1% paraformaldehyde (17). We used live, non-chemically-fixed promastigotes for surface plasmon resonance imaging (SPRi) binding assays in order to preserve their in vivo molecular recognition properties as much as possible.…”

Section: Parasitesmentioning

confidence: 99%

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Large-Scale Investigation of Leishmania Interaction Networks with Host Extracellular Matrix by Surface Plasmon Resonance Imaging

et al. 2014

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We have set up an assay to study the interactions of live pathogens with their hosts by using protein and glycosaminoglycan arrays probed by surface plasmon resonance imaging. We have used this assay to characterize the interactions of Leishmania promastigotes with ϳ70 mammalian host biomolecules (extracellular proteins, glycosaminoglycans, growth factors, cell surface receptors). We have identified, in total, 27 new partners (23 proteins, 4 glycosaminoglycans) of procyclic promastigotes of six Leishmania species and 18 partners (15 proteins, 3 glycosaminoglycans) of three species of stationary-phase promastigotes for all the strains tested. The diversity of the interaction repertoires of Leishmania parasites reflects their dynamic and complex interplay with their mammalian hosts, which depends mostly on the species and strains of Leishmania. Stationary-phase Leishmania parasites target extracellular matrix proteins and glycosaminoglycans, which are highly connected in the extracellular interaction network. Heparin and heparan sulfate bind to most Leishmania strains tested, and 6-O-sulfate groups play a crucial role in these interactions. Numerous Leishmania strains bind to tropoelastin, and some strains are even able to degrade it. Several strains interact with collagen VI, which is expressed by macrophages. Most Leishmania promastigotes interact with several regulators of angiogenesis, including antiangiogenic factors (endostatin, anastellin) and proangiogenic factors (ECM-1, VEGF, and TEM8 [also known as anthrax toxin receptor 1]), which are regulated by hypoxia. Since hypoxia modulates the infection of macrophages by the parasites, these interactions might influence the infection of host cells by Leishmania.

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Section: Discussionsupporting

confidence: 92%

Section: Discussionmentioning

confidence: 99%

Section: Parasitesmentioning

confidence: 99%

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Large-Scale Investigation of Leishmania Interaction Networks with Host Extracellular Matrix by Surface Plasmon Resonance Imaging

et al. 2014

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“…In this regard, it was found the presence of HBP on L. braziliensis promastigotes, suggesting a role of HBP in the interaction of the parasite with the intestinal cells of the insect (Lutzomyia) [15][16][17].…”

Section: Commentarymentioning

confidence: 93%

Parasite Lectins: More than Adhesion Molecules?

Silva¹,

Martins²

2016

J Glycobiol

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CommentarySeveral studies demonstrate the involvement of lectins in the recognition of carbohydrates present on the surface of different kinds of pathogens [1][2][3], including parasites of the genus Leishmania [4,5].The recognition often involves the modulation of the host immune response due to the activation of signaling pathways initiated by the stimulation of lectins present on the surface of cells from immune system by carbohydrates from pathogens [1,2]. On the other hand, lectins from microorganisms are also involved in infections: HeparinBinding Protein (HBP) from Trypanosoma cruzi are involved in processes of amastigote and epimastigotes adhesion to the mammalian host cells and to the intestinal epithelium of insects, respectively [6][7][8]. Furthermore, galactose/N-acetylgalactosamine (Gal/GalNAc) lectin is involved in the infection by Entamoeba histolytica, responsible for the third highest number of death from parasitic diseases in the world [9][10][11]. Other relevant lectins from parasites are involved in mediating protozoa attachment to the host cells, acting as valuable tools to study pathogenesis of infection: mannose lectin (MBP) from Acanthamoeba, causative agent of keratitis, mediates parasite adhesion to the host cells, and may serve as a marker of pathogenicity of this parasite; micronemal protein (MIC1), a Toxoplasma gondii adhesin, bind to host sialic acid moieties, playing role in the parasite invasion and virulence; Tritrichomonas foetus lectin (TFL), a sialic acid specific lectin, is involved in mucosal surface attachment and immunogenicity of Tritrichomonas foetus, a protozoan parasite of the bovine urogenital tract; and Cryptosporidium parvum Clec (CpClec), a novel mucin-like glycoprotein with a C-type lectin domain (CTLD), is involved in Cryptosporidium-host cell interactions [12].Unlike what is observed for lectins present in the vertebrate host, plants or microorganisms, the number of works referring to lectins expression on the surface of Leishmania parasites are limited [13,14], especially with regard to the role of these molecules in cell adhesion and their influence on host immune response. In this regard, it was found the presence of HBP on L. braziliensis promastigotes, suggesting a role of HBP in the interaction of the parasite with the intestinal cells of the insect (Lutzomyia) [15][16][17].Recently, our research group showed the presence of HBP in promastigotes of L. infantum chagasi using Fast Protein Liquid Chromatography (FPLC) with heparin column, providing the basis for further studies on the biological actions of HBP in visceral infection [18]. In this study, HBP was identified on the external membrane of the parasite and also present in the cytoplasm and cytoskeleton, in vesicles, and close to the kinetoplast of the parasite. Furthermore, blocking of HBP from L. infantum chagasi by heparin partially prevented the adhesion and internalization of parasites in RAW 264.7 macrophages; however, it is possible that parasite lectins not only participate in adhesion, as the...

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“…The bound proteins were eluted from the column using the equilibrium buffer containing of 2.0 M NaCl, as previously described [27]. The eluted fractions were concentrated using a Centriprep filter device, and the protein concentration was determined as previously described [28].…”

Section: Methodsmentioning

confidence: 99%

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

et al. 2012

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BackgroundLeishmania (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.MethodsFlagellar (Ff) 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.ResultsThe 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 20μg/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.ConclusionsThe 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.

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Leishmania (Viannia) braziliensis: insights on subcellular distribution and biochemical properties of heparin-binding proteins

Cited by 21 publications

References 51 publications

Large-Scale Investigation of Leishmania Interaction Networks with Host Extracellular Matrix by Surface Plasmon Resonance Imaging

Large-Scale Investigation of Leishmania Interaction Networks with Host Extracellular Matrix by Surface Plasmon Resonance Imaging

Parasite Lectins: More than Adhesion Molecules?

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

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