Mapping of the antigenic determinants of the<i>T. cruzi</i>kinetoplastid membrane protein-11. Identification of a linear epitope specifically recognized by human Chagasic sera

Thomas, M. Carmen; Longobardo, Victoria; Carmelo, Emma; Marañón, Concepción; Planelles, Lourdes; Patarroyo, Manuel E.; Alonso, Carlos; López, Manuel Carlos

doi:10.1046/j.1365-2249.2001.01478.x

Cited by 31 publications

(29 citation statements)

References 23 publications

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“…Purified recombinant T. cruzi KMP-11 (KMP-11r) was obtained as previously described (40,41), reconstituted in 0.13 PBS (Eurobio) and stored at 220˚C.…”

Section: T Cruzi Kmp-11 Recombinant Proteinmentioning

confidence: 99%

Inhibitory Receptor Expression on CD8+ T Cells Is Linked to Functional Responses against Trypanosoma cruzi Antigens in Chronic Chagasic Patients

Lasso

Mateus

Pavía

et al. 2015

The Journal of Immunology

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In mammals, chronic diseases resulting from infectious agents have been associated with functional T cell response deficiency, a high frequency of terminally differentiated T cells, the presence of monofunctional Ag-specific T cells, and increased expression of inhibitory receptors. Similar to other chronic diseases, the progressive loss of certain functional activities during Trypanosoma cruzi infection might result in the inability to control replication of this parasite. To examine this hypothesis, we evaluated the differentiation and cell effector function of CD8+ T cells and characterized the expression of inhibitory receptors and the presence of the parasite in the bloodstream of chagasic patients. The results showed that patients at an advanced severe disease stage had a higher frequency of terminally differentiated CD8+ T cells than patients at an early stage of the disease. A monofunctional CD8+ T cell response was observed in patients at an advanced stage, whereas the coexpression of markers that perform three and four functions in response to parasite Ags was observed in patients at a less severe disease stage. The frequency of CD8+ T cells producing granzyme B and perforin and those expressing inhibitory receptors was higher in symptomatic patients than in asymptomatic patients. Taken together, these findings suggest that during the course of Chagas disease, CD8+ T cells undergo a gradual loss of function characterized by impaired cytokine production, the presence of advanced differentiation, and increased inhibitory receptor coexpression.

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“…Purified recombinant T. cruzi KMP-11 (KMP-11r) was obtained as previously described (40,41), reconstituted in 0.13 PBS (Eurobio) and stored at 220˚C.…”

Section: T Cruzi Kmp-11 Recombinant Proteinmentioning

confidence: 99%

Inhibitory Receptor Expression on CD8+ T Cells Is Linked to Functional Responses against Trypanosoma cruzi Antigens in Chronic Chagasic Patients

Lasso

Mateus

Pavía

et al. 2015

The Journal of Immunology

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“…As a control, we immunized mice with the empty vector or with saline solution. The anti-KMP11-specific antibody levels were determined by enzyme-linked immunosorbent assay (ELISA) using purified KMP11 recombinant protein as an antigen, obtained as previously described (29). The antibody response (immunoglobulin G [IgG]) induced by the DNA constructs is shown in Fig.…”

mentioning

confidence: 99%

DNA Immunization withTrypanosoma cruziHSP70 Fused to the KMP11 Protein Elicits a Cytotoxic and Humoral Immune Response against the Antigen and Leads to Protection

et al. 2001

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Murine immunization with Trypanosoma cruzi KMP11-HSP70 fused genes but not the KMP11 gene alone elicited both an immunoglobulin G2a long-lasting humoral immune response against KMP11 protein and activation of CD8 ؉ cytotoxic T lymphocytes specific for two KMP11 peptides containing A2 motifs. Moreover, protection against the parasite challenge was observed after immunization with the chimeric gene.

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“…The latter epitope has been shown to react specifically with Ͼ50% of T. cruzi sera when chemically immobilized on a 96-well plate (33,35). By including the Kmp-11 sequence in the multiepitope recombinant, it was not subject to the need for such immobilization.…”

Section: Resultsmentioning

confidence: 99%

Lateral Flow Immunoassay for Diagnosis of Trypanosoma cruzi Infection with High Correlation to the Radioimmunoprecipitation Assay

Houghton

Stevens

Hjerrild

et al. 2009

Clin Vaccine Immunol

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The incidence of blood donors seropositive for Trypanosoma cruzi in North America has increased with population migration and more rigorous surveillance. The United States, considered nonendemic for T. cruzi, could therefore be at risk to exposure to parasite transmission through blood or organ donations. Current tests show variable reactivity, especially with Central American sera. Here we describe the development of a lateral flow immunoassay for the rapid detection of T. cruzi infection that has a strong correlation to the radioimmunoprecipitation assay (RIPA) "gold standard" in the United States. Such a test could have utility in small blood banks for prescreening donors, as well as in cardiac transplantation evaluation. T. cruzi consensus and/or RIPA-positive sera from Central and South America were evaluated in enzyme immunoassays (EIAs). These included commercial panels from Boston Biomedica, Inc. (BBI) (n ‫؍‬ 14), and HemaBio (n ‫؍‬ 21). Other sources included RIPA-positive sera from the American Red Cross (ARC) (n ‫؍‬ 42), as well as from Chile. Sera were tested with the multiepitope recombinant TcF. All but one of the BBI samples were positive and 7 of 21 HemaBio samples and 6 of 42 ARC samples were low positive or negative. This observation indicated the need for additional antigens. To complement TcF reactivity, we tested the sera with peptides 30, 36, SAPA, and 1.1, 1.2, and 1.3 His fragments of 85-kDa trans-sialidase. We identified a promising combination of the tested antigens and constructed a single recombinant protein, ITC6, that enhanced the relative sensitivity in U.S. blood donor sera compared to that of TcF. The data on its evaluation using RIPA-confirmed positive sera in EIA and lateral flow immunoassay studies are presented, along with an additional recombinant protein, ITC8.2, with two additional sequences for peptide 1 and Kmp-11. The latter, when evaluated in a dipstick assay with consensus positive sera, had a sensitivity of 99.2% and a specificity of 99.1%.

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Mapping of the antigenic determinants of theT. cruzikinetoplastid membrane protein-11. Identification of a linear epitope specifically recognized by human Chagasic sera

Cited by 31 publications

References 23 publications

Inhibitory Receptor Expression on CD8+ T Cells Is Linked to Functional Responses against Trypanosoma cruzi Antigens in Chronic Chagasic Patients

Inhibitory Receptor Expression on CD8+ T Cells Is Linked to Functional Responses against Trypanosoma cruzi Antigens in Chronic Chagasic Patients

DNA Immunization withTrypanosoma cruziHSP70 Fused to the KMP11 Protein Elicits a Cytotoxic and Humoral Immune Response against the Antigen and Leads to Protection

Lateral Flow Immunoassay for Diagnosis of Trypanosoma cruzi Infection with High Correlation to the Radioimmunoprecipitation Assay

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