Methodological approach to the ex vivo expansion and detection of T. cruzi-specific T cells from chronic Chagas disease patients
| Autor: | Radleigh G. Santos, Augusto Atienza, Nazila Sabri, María Paola Zago, Silvia Andrea Longhi, Karina A. Gómez, Alcinette Bunying, Gonzalo Raúl Acevedo, Clemencia Pinilla, Valeria Judkowski |
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| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
T Cell Library
0301 basic medicine Pathology Physiology lcsh:Medicine medicine.disease_cause Memory T cells purl.org/becyt/ford/1 [https] White Blood Cells 0302 clinical medicine Cognition Learning and Memory Animal Cells Cellular types Medicine and Health Sciences Medicine lcsh:Science Protozoans Trypanosoma Cruzi Multidisciplinary biology 3. Good health Immunologic memory CIENCIAS NATURALES Y EXACTAS Research Article Neglected Tropical Diseases Chagas disease medicine.medical_specialty Trypanosoma Cell biology Blood cells Immune Cells Immunology Chronic Chagas' disease T cells Antigen-Presenting Cells Ciencias Biológicas Memory T Cells 03 medical and health sciences Biología Celular Microbiología Memory Parasitic Diseases Chagas Disease Ex vivo expansion purl.org/becyt/ford/1.6 [https] Antigen-presenting cell Trypanosoma cruzi Secretion Protozoan Infections Biology and life sciences business.industry lcsh:R Organisms medicine.disease biology.organism_classification Tropical Diseases Virology Epstein–Barr virus Parasitic Protozoans 030104 developmental biology Chronic disease Cognitive Science lcsh:Q business Physiological Processes 030215 immunology Neuroscience |
| Zdroj: | PLoS ONE CONICET Digital (CONICET) Consejo Nacional de Investigaciones Científicas y Técnicas instacron:CONICET PLoS ONE, Vol 12, Iss 5, p e0178380 (2017) |
| ISSN: | 1932-6203 |
| Popis: | The discovery of T cell epitopes is essential not only for gaining knowledge about host response to infectious disease but also for the development of immune-intervention strategies. In Chagas disease, given the size and complexity of the Trypanosoma cruzi proteome and its interaction with the host’s immune system, the fine specificity of T cells has not been extensively studied yet, and this is particularly true for the CD4+ T cell compartment. The aim of the present work was to optimize a protocol for the generation of parasite-specific memory T cell lines, representative of their in vivo precursor populations and capable of responding to parasite antigens after long-term culture. Accordingly, peripheral blood mononuclear cells (PBMC) from both chronic asymptomatic and cardiac patients, and from non-infected individuals, underwent different in vitro culture and stimulation conditions. Subsequently, cells were tested for their capacity to respond against T. cruzi lysate by measuring [3H]-thymidine incorporation and interferon-γ and GM-CSF secretion. Results allowed us to adjust initial T. cruzi lysate incubation time as well as the number of expansions with phytohemagglutinin (PHA) and irradiated allogeneic PBMC prior to specificity evaluation. Moreover, our data demonstrated that parasite specific T cells displayed a clear and strong activation by using T. cruzi lysate pulsed, Epstein-Barr virus (EBV)-transformed human B lymphocytes (B-LCL), as autologous antigen presenting cells. Under these culture conditions, we generated a clone from an asymptomatic patient’s memory CD4+ T cells which responded against epimastigote and trypomastigote protein lysate. Our results describe a culture method for isolating T. cruzi specific T cell clones from patients with Chagas disease, which enable the acquisition of information on functionality and specificity of individual T cells. Fil: Acevedo, Gonzalo Raúl. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina Fil: Longhi, Silvia Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina Fil: Bunying, Alcinette. Torrey Pines Institute for Molecular Studies; Estados Unidos Fil: Sabri, Nazila. Torrey Pines Institute for Molecular Studies; Estados Unidos Fil: Atienza, Augusto. Gobierno de la Ciudad de Buenos Aires. Hospital General de Agudos ; Argentina Fil: Zago, María Paola. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; Argentina Fil: Santos, Radleigh. Torrey Pines Institute for Molecular Studies; Estados Unidos Fil: Judkowski, Valeria A.. Torrey Pines Institute for Molecular Studies; Estados Unidos Fil: Pinilla, Clemencia. Torrey Pines Institute for Molecular Studies; Estados Unidos Fil: Gomez, Karina Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina |
| Databáze: | OpenAIRE |
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