TcTASV-C, a Protein Family in Trypanosoma cruzi that Is Predominantly Trypomastigote-Stage Specific and Secreted to the Medium

Bernabó, Guillermo; Levy, Gabriela Vanesa; Ziliani, María; Caeiro, Lucas; Sánchez, Daniel O.; Tekiel, Valeria

doi:10.1371/journal.pone.0071192

Cited by 23 publications

(52 citation statements)

References 59 publications

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“…The puzzling non-colocalizing patterns found for the different proteins analyzed, together with other reports of trypomastigote proteins that also displayed dotted patterns [ 18 , 25 , 48 ] suggest a particular membrane topology with defined domains. To obtain highly solved topological information, we then assayed atomic force microscopy (AFM), a suitable tool that has been used in epimastigotes [ 49 , 50 ].…”

Section: Resultsmentioning

confidence: 59%

“…Despite their complexity and variations in amino acid sequence, TcMUC II deduced products share a common structure made up of a highly conserved N -terminal signal peptide, a variable and central region showing highly biased amino acid composition, in which Thr, Ser, Pro, Gly, and Ala residues together might add up to 60–80% of the total count, and a C -terminal GPI attachment signal. Besides the TcMUC genes, other families of glycosylated proteins families such as TcTASV [ 18 ] and MASP [ 7 ] are being discovered. To date, the structure of the O -linked oligosaccharides has only been determined for the mucins from the non-infective insect epimastigote stage (known as Gp35/50 mucins) of different strains of T .…”

Section: Introductionmentioning

confidence: 99%

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Sialic Acid Glycobiology Unveils Trypanosoma cruzi Trypomastigote Membrane Physiology

et al. 2016

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Trypanosoma cruzi, the flagellate protozoan agent of Chagas disease or American trypanosomiasis, is unable to synthesize sialic acids de novo. Mucins and trans-sialidase (TS) are substrate and enzyme, respectively, of the glycobiological system that scavenges sialic acid from the host in a crucial interplay for T. cruzi life cycle. The acquisition of the sialyl residue allows the parasite to avoid lysis by serum factors and to interact with the host cell. A major drawback to studying the sialylation kinetics and turnover of the trypomastigote glycoconjugates is the difficulty to identify and follow the recently acquired sialyl residues. To tackle this issue, we followed an unnatural sugar approach as bioorthogonal chemical reporters, where the use of azidosialyl residues allowed identifying the acquired sugar. Advanced microscopy techniques, together with biochemical methods, were used to study the trypomastigote membrane from its glycobiological perspective. Main sialyl acceptors were identified as mucins by biochemical procedures and protein markers. Together with determining their shedding and turnover rates, we also report that several membrane proteins, including TS and its substrates, both glycosylphosphatidylinositol-anchored proteins, are separately distributed on parasite surface and contained in different and highly stable membrane microdomains. Notably, labeling for α(1,3)Galactosyl residues only partially colocalize with sialylated mucins, indicating that two species of glycosylated mucins do exist, which are segregated at the parasite surface. Moreover, sialylated mucins were included in lipid-raft-domains, whereas TS molecules are not. The location of the surface-anchored TS resulted too far off as to be capable to sialylate mucins, a role played by the shed TS instead. Phosphatidylinositol-phospholipase-C activity is actually not present in trypomastigotes. Therefore, shedding of TS occurs via microvesicles instead of as a fully soluble form.

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Section: Resultsmentioning

confidence: 59%

Section: Introductionmentioning

confidence: 99%

Sialic Acid Glycobiology Unveils Trypanosoma cruzi Trypomastigote Membrane Physiology

et al. 2016

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“…Although a vesicular location near the flagellar pocket has been described in trypomastigotes [ 6 ], its location on the surface cannot be ruled out since many T . cruzi protein has been shown to be temporary located on the membrane [ 45 – 47 ].…”

Section: Discussionmentioning

confidence: 99%

Trypanosoma cruzi 80 kDa prolyl oligopeptidase (Tc80) as a novel immunogen for Chagas disease vaccine

et al. 2018

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BackgroundChagas disease, also known as American Trypanosomiasis, is a chronic parasitic disease caused by the flagellated protozoan Trypanosoma cruzi that affects about 8 million people around the world where more than 25 million are at risk of contracting the infection. Despite of being endemic on 21 Latin-American countries, Chagas disease has become a global concern due to migratory movements. Unfortunately, available drugs for the treatment have several limitations and they are generally administered during the chronic phase of the infection, when its efficacy is considered controversial. Thus, prophylactic and/or therapeutic vaccines are emerging as interesting control alternatives. In this work, we proposed Trypanosoma cruzi 80 kDa prolyl oligopeptidase (Tc80) as a new antigen for vaccine development against Chagas disease.Methodology/Principal findingsIn a murine model, we analyzed the immune response triggered by different immunization protocols based on Tc80 and evaluated their ability to confer protection against a challenge with the parasite. Immunized mice developed Tc80-specific antibodies which were able to carry out different functions such as: enzymatic inhibition, neutralization of parasite infection and complement-mediated lysis of trypomastigotes. Furthermore, vaccinated mice elicited strong cell-mediated immunity. Spleen cells from immunized mice proliferated and secreted Th1 cytokines (IL-2, IFN-γ and TNF-α) upon re-stimulation with rTc80. Moreover, we found Tc80-specific polyfunctional CD4 T cells, and cytotoxic T lymphocyte activity against one Tc80 MHC-I peptide.Immunization protocols conferred protection against a T. cruzi lethal challenge. Immunized groups showed a decreased parasitemia and higher survival rate compared with non-immunized control mice. Moreover, during the chronic phase of the infection, immunized mice presented: lower levels of myopathy-linked enzymes, parasite burden, electrocardiographic disorders and inflammatory cells.Conclusions/SignificanceConsidering that an early control of parasite burden and tissue damage might contribute to avoid the progression towards symptomatic forms of chronic Chagas disease, the efficacy of Tc80-based vaccines make this molecule a promising immunogen for a mono or multicomponent vaccine against T. cruzi infection.

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“…Peptides from the same protein were synthesized at randomized positions within a single sector. The chip contains additional 2386 unique peptides corresponding to eight proteins not accounted within the protein groups defined above (and of which 2 proteins resulted seropositive), including members of the T. cruzi TASV protein family ( 30 , 31 ) and 21 epitopes of high prevalence in healthy humans (not associated to Chagas disease, identified in the IEDB with a search for linear B-cell epitopes from any source with sero-prevalence > = 50%, assayed in at least 20 subjects). The sequences of these 532 proteins (457 T. cruzi proteins, 54 neo-proteins and the additional 21 short peptidic epitopes of high prevalence in healthy humans) are available in supplemental Table S4 .…”

Section: Methodsmentioning

confidence: 99%

Towards High-throughput Immunomics for Infectious Diseases: Use of Next-generation Peptide Microarrays for Rapid Discovery and Mapping of Antigenic Determinants

Carmona

Nielsen

Schafer‐Nielsen

et al. 2015

Molecular & Cellular Proteomics

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101

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Complete characterization of antibody specificities associated to natural infections is expected to provide a rich source of serologic biomarkers with potential applications in molecular diagnosis, follow-up of chemotherapeutic treatments, and prioritization of targets for vaccine development. Here, we developed a highly-multiplexed platform based on next-generation high-density peptide microarrays to map these specificities in Chagas Disease, an exemplar of a human infectious disease caused by the protozoan Trypanosoma cruzi. We designed a high-density peptide microarray containing more than 175,000 overlapping 15mer peptides derived from T. cruzi proteins. Peptides were synthesized in situ on microarray slides, spanning the complete length of 457 parasite proteins with fully overlapped 15mers (1 residue shift). Screening of these slides with antibodies purified from infected patients and healthy donors demonstrated both a high technical reproducibility as well as epitope mapping consistency when compared with earlier low-throughput technologies. Using a conservative signal threshold to classify positive (reactive) peptides we identified 2,031 disease-specific peptides and 97 novel parasite antigens, effectively doubling the number of known antigens and providing a 10-fold increase in the number of fine mapped antigenic determinants for this disease. Finally, further analysis of the chip data showed that optimizing the amount of sequence overlap of displayed peptides can increase the protein space covered in a single chip by at least ∼threefold without sacrificing sensitivity. In conclusion, we show the power of high-density peptide chips for the discovery of pathogen-specific linear B-cell epitopes from clinical samples, thus setting the stage for high-throughput biomarker discovery screenings and proteome-wide studies of immune responses against pathogens.

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TcTASV-C, a Protein Family in Trypanosoma cruzi that Is Predominantly Trypomastigote-Stage Specific and Secreted to the Medium

Cited by 23 publications

References 59 publications

Sialic Acid Glycobiology Unveils Trypanosoma cruzi Trypomastigote Membrane Physiology

Sialic Acid Glycobiology Unveils Trypanosoma cruzi Trypomastigote Membrane Physiology

Trypanosoma cruzi 80 kDa prolyl oligopeptidase (Tc80) as a novel immunogen for Chagas disease vaccine

Towards High-throughput Immunomics for Infectious Diseases: Use of Next-generation Peptide Microarrays for Rapid Discovery and Mapping of Antigenic Determinants

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