Molecular and antigenic characterization of Trypanosoma cruzi TolT proteins

Lobo, Mabel Maite; Balouz, Virginia; Melli, Luciano Jorge; Carlevaro, Giannina; Cortina, María Eugenia; Camara, María de los Milagros; Canepa, Gaspar E.; Carmona, Santiago J.; Altcheh, Jaime; Campetella, Oscar; Ciocchini, Andrés E.; Agüero, Fernán; Mucci, Juan; Buscaglia, Carlos A.

doi:10.1371/journal.pntd.0007245

Cited by 6 publications

(6 citation statements)

References 65 publications

(105 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…As expected, Tc964 was recognized by sera from CCD patients, confirming its antigenic properties, as previously reported for other kinetoplastid conserved proteins, such as the Kinetoplastid membrane protein-11 (KMP-11) [57] and TolT (TolA-like protein) proteins from T. cruzi [47]. Interestingly, sera'from the B group of patients, who in spite of being asymptomatic already present structural heart damage, showed the lowest reactivity ( Figure 4C).…”

Section: Discussionsupporting

confidence: 87%

“…The characterization of proteins from pathogens of medical importance, as T. cruzi, is relevant because it provides information on different biological aspects of the parasite and allows assessing properties of these proteins to define them as targets for developing disease intervention strategies [47]. Based on this rationale, herein we undertook the molecular characterization of a novel T. cruzi protein, Tc964.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Molecular Characterization of Tc964, A Novel Antigenic Protein from Trypanosoma cruzi

Ruiz-Márvez

Ramírez

Rodríguez

et al. 2020

IJMS

View full text Add to dashboard Cite

The Tc964 protein was initially identified by its presence in the interactome associated with the LYT1 mRNAs, which code for a virulence factor of Trypanosoma cruzi. Tc964 is annotated in the T. cruzi genome as a hypothetical protein. According to phylogenetic analysis, the protein is conserved in the different genera of the Trypanosomatidae family; however, recognizable orthologues were not identified in other groups of organisms. Therefore, as a first step, an in-depth molecular characterization of the Tc946 protein was carried out. Based on structural predictions and molecular dynamics studies, the Tc964 protein would belong to a particular class of GTPases. Subcellular fractionation analysis indicated that Tc964 is a nucleocytoplasmic protein. Additionally, the protein was expressed as a recombinant protein in order to analyze its antigenicity with sera from Chagas disease (CD) patients. Tc964 was found to be antigenic, and B-cell epitopes were mapped by the use of synthetic peptides. In parallel, the Leishmania major homologue (Lm964) was also expressed as recombinant protein and used for a preliminary evaluation of antigen cross-reactivity in CD patients. Interestingly, Tc964 was recognized by sera from Chronic CD (CCD) patients at different stages of disease severity, but no reactivity against this protein was observed when sera from Colombian patients with cutaneous leishmaniasis were analyzed. Therefore, Tc964 would be adequate for CD diagnosis in areas where both infections (CD and leishmaniasis) coexist, even though additional assays using larger collections of sera are needed in order to confirm its usefulness for differential serodiagnosis.

show abstract

Section: Discussionsupporting

confidence: 87%

Section: Discussionmentioning

confidence: 99%

Molecular Characterization of Tc964, A Novel Antigenic Protein from Trypanosoma cruzi

Ruiz-Márvez

Ramírez

Rodríguez

et al. 2020

IJMS

View full text Add to dashboard Cite

show abstract

“…No T. cruzi antigens could be detected by western blotting in EV-THP-1 and the presence of plasma membrane glycoproteins expressing α-Gal epitopes (i.e., TcMUCII mucins or tGPI-mucins) of the parasite in the EVs of macrophages infected by T. cruzi were not identified by total proteomic analysis. In this analysis only six parasite proteins were found in EVs from macrophages infected with T. cruzi i.e., HSP60, tryparedoxin peroxidase, trans-sialidase Group II, flagellar calcium-binding protein, surface protein TolT and paraflagellar rod protein 2 (Lobo et al, 2019). These parasite proteins are involved in the recognition of the parasite by host cells, invasion, the adhesion of host cells, metabolism and the induction of the host immune response (Godsel et al, 1995;Quanquin et al, 1999;Mattos et al, 2014;Urményi et al, 2014;Girard et al, 2018).…”

Section: Discussionmentioning

confidence: 92%

Trypanosoma cruzi-Infected Human Macrophages Shed Proinflammatory Extracellular Vesicles That Enhance Host-Cell Invasion via Toll-Like Receptor 2

Andrade

Xander

Soares

et al. 2020

Front. Cell. Infect. Microbiol.

View full text Add to dashboard Cite

Extracellular vesicles (EVs) shed by trypomastigote forms of Trypanosoma cruzi have the ability to interact with host tissues, increase invasion, and modulate the host innate response. In this study, EVs shed from T. cruzi or T.cruzi-infected macrophages were investigated as immunomodulatory agents during the initial steps of infection. Initially, by scanning electron microscopy and nanoparticle tracking analysis, we determined that T. cruzi-infected macrophages release higher numbers of EVs (50-300 nm) as compared to non-infected cells. Using Toll-like-receptor 2 (TLR2)-transfected CHO cells, we observed that pre-incubation of these host cells with parasite-derived EVs led to an increase in the percentage of infected cells. In addition, EVs from parasite or T.cruzi-infected macrophages or not were able to elicit translocation of NF-κB by interacting with TLR2, and as a consequence, to alter the EVs the gene expression of proinflammatory cytokines (TNF-α, IL-6, and IL-1β), and STAT-1 and STAT-3 signaling pathways. By proteomic analysis, we observed highly significant changes in the protein composition between non-infected and infected host cell-derived EVs. Thus, we observed the potential of EVs derived from T. cruzi during infection to maintain the inflammatory response in the host.

show abstract

“…Variations on this theme, aimed at improving the exposition of peptidic ligand-binding domains of surface molecules by means of heavily glycosylated regions have been proposed for TcSMUG L glycoproteins [6] as well as for other T . cruzi surface molecules [16,69] and a multiplicity of yeast flocculins and adhesins [70].…”

Section: Discussionmentioning

confidence: 99%

Trypanosoma cruzi surface mucins are involved in the attachment to the Triatoma infestans rectal ampoule

et al. 2019

Self Cite

View full text Add to dashboard Cite

Background Trypanosoma cruzi , the agent of Chagas disease, is a protozoan parasite transmitted to humans by blood-sucking triatomine vectors. However, and despite its utmost biological and epidemiological relevance, T . cruzi development inside the digestive tract of the insect remains a poorly understood process. Methods/Principle findings Here we showed that Gp35/50 kDa mucins, the major surface glycoproteins from T . cruzi insect-dwelling forms, are involved in parasite attachment to the internal cuticle of the triatomine rectal ampoule, a critical step leading to its differentiation into mammal-infective forms. Experimental evidence supporting this conclusion could be summarized as follows: i) native and recombinant Gp35/50 kDa mucins directly interacted with hindgut tissues from Triatoma infestans , as assessed by indirect immunofluorescence assays; ii) transgenic epimastigotes over-expressing Gp35/50 kDa mucins on their surface coat exhibited improved attachment rates (~2–3 fold) to such tissues as compared to appropriate transgenic controls and/or wild-type counterparts; and iii) certain chemically synthesized compounds derived from Gp35/50 kDa mucins were able to specifically interfere with epimastigote attachment to the inner lining of T . infestans rectal ampoules in ex vivo binding assays, most likely by competing with or directly blocking insect receptor(s). A solvent-exposed peptide (smugS peptide) from the Gp35/50 kDa mucins protein scaffolds and a branched, Gal f -containing trisaccharide (Gal f β1–4[Gal p β1–6]GlcNAcα) from their O -linked glycans were identified as main adhesion determinants for these molecules. Interestingly, exogenous addition of a synthetic Gal f β1–4[Gal p β1–6]GlcNAcα derivative or of oligosaccharides containing this structure impaired the attachment of Dm28c but not of CL Brener epimastigotes to triatomine hindgut tissues; which correlates with the presence of Gal f residues on the Gp35/50 kDa mucins’ O -glycans on the former but not the latter parasite clone. Conclusion/Significance These results provide novel insights into the mechanisms underlying T . cruzi- triatomine interplay, and indicate that inter-strain variations in the O -glycosylation of Gp35/50 kDa mucins may lead to differences in parasite differentiation and hence, in parasite transmissibility to the mammalian host. Most importantly, our findings point to Gp35/50 kDa mucins and/or the Gal f ...

show abstract

Molecular and antigenic characterization of Trypanosoma cruzi TolT proteins

Cited by 6 publications

References 65 publications

Molecular Characterization of Tc964, A Novel Antigenic Protein from Trypanosoma cruzi

Molecular Characterization of Tc964, A Novel Antigenic Protein from Trypanosoma cruzi

Trypanosoma cruzi-Infected Human Macrophages Shed Proinflammatory Extracellular Vesicles That Enhance Host-Cell Invasion via Toll-Like Receptor 2

Trypanosoma cruzi surface mucins are involved in the attachment to the Triatoma infestans rectal ampoule

Contact Info

Product

Resources

About