cDNA cloning and partial characterization of amastigote specific surface protein from Trypanosoma cruzi

Olivas-Rubio, Marybell; Hernández‐Martínez, Salvador; Talamás-Rohana, Patricia; Tsutsumi, Vı́ctor; Reyes-López, Pedro Antonio; Rosales‐Encina, José Luis

doi:10.1016/j.meegid.2009.05.016

Cited by 5 publications

(7 citation statements)

References 56 publications

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“…The gene encoding Tc SSP4 (amastigote-specific protein) was obtained from the pBluescriptSk-Am230 plasmid [21] as a 2.2 kb Eco RI (Boehringer Mannheim) fragment. The gene encoding Tc SP ( trans -sialidase protein) was released from pBluescriptSk-A83 plasmid (Rosales-Encina JL, unpublished observations) as a 6.0 kb Eco RI fragment.…”

Section: Methodsmentioning

confidence: 99%

Plasmid DNA immunization with Trypanosoma cruzi genes induces cardiac and clinical protection against Chagas disease in the canine model

Rodríguez-Morales

Pérez-Leyva

Ballinas-Verdugo

et al. 2012

Vet Res

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The only existing preventive measure against American trypanosomosis, or Chagas disease, is the control of the transmitting insect, which has only been effective in a few South American regions. Currently, there is no vaccine available to prevent this disease. Here, we present the clinical and cardiac levels of protection induced by expression to Trypanosoma cruzi genes encoding the TcSP and TcSSP4 proteins in the canine model. Physical examination, diagnostic chagasic serology, and serial electrocardiograms were performed before and after immunization, as well as after experimental infection. We found that immunization with recombinant plasmids prevented hyperthermia in the acute phase of experimental infection and produced lymphadenomegaly as an immunological response against the parasite and additionally prevented heart rate elevation (tachycardia) in the acute and/or chronic stages of infection. Immunization with T. cruzi genes encoding the TcSP and TcSSP4 antigens diminished the quality and quantity of the electrocardiographic abnormalities, thereby avoiding progression to more severe developments such as right bundle branch block or ventricular premature complexes in a greater number of dogs.

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Section: Methodsmentioning

confidence: 99%

Plasmid DNA immunization with Trypanosoma cruzi genes induces cardiac and clinical protection against Chagas disease in the canine model

Rodríguez-Morales

Pérez-Leyva

Ballinas-Verdugo

et al. 2012

Vet Res

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“…The first description of Ssp-4 revealed that mAb2C2 recognizes a surface GPI-anchored glycoprotein, which is cleaved and released into extracellular milieu in the presence of PI-PLC ( Andrews et al, 1988 ). Subsequently, Olivas-Rubio et al (2009) identified a partial sequence from the DGF-1 multigenic family, but identification of potential domains on the putative protein was challenging. These challenges led us to identify Ssp-4 using approaches other than immunostaining, such as mass spectrometry (MS) of mAb2C2- and mAb1D9-immunoprecipitated proteins, protein sequencing with bioinformatic analysis and quantitative PCR.…”

Section: Discussionmentioning

confidence: 99%

A Carbohydrate Moiety of Secreted Stage-Specific Glycoprotein 4 Participates in Host Cell Invasion by Trypanosoma cruzi Extracellular Amastigotes

et al. 2018

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Trypanosoma cruzi is the etiologic agent of Chagas’ disease. It is known that amastigotes derived from trypomastigotes in the extracellular milieu are infective in vitro and in vivo. Extracellular amastigotes (EAs) have a stage-specific surface antigen called Ssp-4, a GPI-anchored glycoprotein that is secreted by the parasites. By immunoprecipitation with the Ssp-4-specific monoclonal antibodies (mAb) 2C2 and 1D9, we isolated the glycoprotein from EAs. By mass spectrometry, we identified the core protein of Ssp-4 and evaluated mRNA expression and the presence of Ssp-4 carbohydrate epitopes recognized by mAb1D9. We demonstrated that the carbohydrate epitope recognized by mAb1D9 could promote host cell invasion by EAs. Although infectious EAs express lower amounts of Ssp-4 compared with less-infectious EAs (at the mRNA and protein levels), it is the glycosylation of Ssp-4 (identified by mAb1D9 staining only in infectious strains and recognized by galectin-3 on host cells) that is the determinant of EA invasion of host cells. Furthermore, Ssp-4 is secreted by EAs, either free or associated with parasite vesicles, and can participate in host-cell interactions. The results presented here describe the possible role of a carbohydrate moiety of T. cruzi surface glycoproteins in host cell invasion by EA forms, highlighting the potential of these moieties as therapeutic and vaccine targets for the treatment of Chagas’ disease.

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“…At the time of their first encounter with their host, parasites might modulate the immune response by actively secreting or expressing molecules with potent effects on the immune system [8, 18]. A large variety of modulatory parasite-derived proteins, lipids, and glycoconjugates has been described [10]. In the T. cruzi protozoan parasite, specifically in the amastigote stage, a surface glycoprotein named SSP4 was described by Andrews et al [9].…”

Section: Discussionmentioning

confidence: 99%

“…E. coli DH5- α was transformed with this plasmid to obtain the fusion protein MBP::SSP4 (rSSP4) [10, 11]. rSSP4 and MBP were purified by amylase affinity chromatography; after purification, material was analyzed by 10% SDS-PAGE on which a 127 kDa protein corresponding to rSSP4 and a 43 kDa protein corresponding to MBP were observed, respectively (data not shown).…”

Section: Methodsmentioning

confidence: 99%

“…Newly transformed amastigotes, both intracellular and extracellular, express a major surface glycoprotein (SSP4) bound to the plasma membrane by a GPI anchor [9]. The gene that codifies for this protein was cloned [10], and rSSP4 was shown to be a modulator of the immune response, inducing high levels of IgG1, IgG2a and IgG2b isotypes, and the expression of iNOS and production of NO by MΦs [11]. Moreover, rSSP4 was also able to induce the mRNA for IL-1 α , IL-6, IL-12, IFN- γ , and TNF- α cytokines in normal mice, and IL-10 in immunized mice [11], suggesting that TcSSP4 may be involved in modulating T cell populations during T. cruzi infection.…”

Section: Introductionmentioning

confidence: 99%

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Treg Cells Induced by rSSP4 Derived fromT. cruziAmastigotes Increase Parasitemia in an Experimental Chagas Disease Model

Flores-García

Rosales‐Encina

Rosales-García

et al. 2013

BioMed Research International

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Currently, there is a considerable controversy over the participation of Treg cells during Trypanosoma cruzi infection, the main point being whether these cells play a negative or a positive role. In this work, we found that the adoptive transfer of CD4+CD25+FOXP3+ T cells from rSSP4- (a recombinant Trypanosoma cruzi amastigote derived protein, previously shown to have immunomodulatory properties on macrophages) immunized BALB/c donors into syngenic recipients simultaneously with T. cruzi challenge reduces cardiac inflammation and prolongs hosts' survival but increases blood parasitemia and parasite loads in the heart. These CD4+CD25+FOXP3+ Treg cells from immunized mice have a relatively TGF-β-dependent suppressive activity on CD4+ T cells. Therefore, regulatory CD4+CD25+ T cells play a positive role in the development of acute T. cruzi infection by inducing immunosuppressive activity that controls early cardiac inflammation during acute Chagas disease, prolonging survival, but at the same time promoting parasite growth.

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cDNA cloning and partial characterization of amastigote specific surface protein from Trypanosoma cruzi

Cited by 5 publications

References 56 publications

Plasmid DNA immunization with Trypanosoma cruzi genes induces cardiac and clinical protection against Chagas disease in the canine model

Plasmid DNA immunization with Trypanosoma cruzi genes induces cardiac and clinical protection against Chagas disease in the canine model

A Carbohydrate Moiety of Secreted Stage-Specific Glycoprotein 4 Participates in Host Cell Invasion by Trypanosoma cruzi Extracellular Amastigotes

Treg Cells Induced by rSSP4 Derived fromT. cruziAmastigotes Increase Parasitemia in an Experimental Chagas Disease Model

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