Methodological approach to the ex vivo expansion and detection of T. cruzi-specific T cells from chronic Chagas disease patients

Autores
Acevedo, Gonzalo Raúl; Longhi, Silvia Andrea; Bunying, Alcinette; Sabri, Nazila; Atienza, Augusto; Zago, María Paola; Santos, Radleigh; Judkowski, Valeria A.; Pinilla, Clemencia; Gomez, Karina Andrea
Año de publicación
2017
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
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
Materia
Chagas Disease
Memory T Cells
Trypanosoma Cruzi
T Cell Library
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/43074
Acceder
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repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Methodological approach to the ex vivo expansion and detection of T. cruzi-specific T cells from chronic Chagas disease patientsAcevedo, Gonzalo RaúlLonghi, Silvia AndreaBunying, AlcinetteSabri, NazilaAtienza, AugustoZago, María PaolaSantos, RadleighJudkowski, Valeria A.Pinilla, ClemenciaGomez, Karina AndreaChagas DiseaseMemory T CellsTrypanosoma CruziT Cell Libraryhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1https://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1The 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"; ArgentinaFil: Longhi, Silvia Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Bunying, Alcinette. Torrey Pines Institute for Molecular Studies; Estados UnidosFil: Sabri, Nazila. Torrey Pines Institute for Molecular Studies; Estados UnidosFil: Atienza, Augusto. Gobierno de la Ciudad de Buenos Aires. Hospital General de Agudos ; ArgentinaFil: 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; ArgentinaFil: Santos, Radleigh. Torrey Pines Institute for Molecular Studies; Estados UnidosFil: Judkowski, Valeria A.. Torrey Pines Institute for Molecular Studies; Estados UnidosFil: Pinilla, Clemencia. Torrey Pines Institute for Molecular Studies; Estados UnidosFil: Gomez, Karina Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaPublic Library of Science2017-05-26info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/43074Acevedo, Gonzalo Raúl; Longhi, Silvia Andrea; Bunying, Alcinette; Sabri, Nazila; Atienza, Augusto; et al.; Methodological approach to the ex vivo expansion and detection of T. cruzi-specific T cells from chronic Chagas disease patients; Public Library of Science; Plos One; 12; 5; 26-5-2017; 1-19; e01844671932-6203CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1371/journal.pone.0178380info:eu-repo/semantics/altIdentifier/url/http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0178380info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-15T14:35:52Zoai:ri.conicet.gov.ar:11336/43074instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-10-15 14:35:52.513CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Methodological approach to the ex vivo expansion and detection of T. cruzi-specific T cells from chronic Chagas disease patients
title Methodological approach to the ex vivo expansion and detection of T. cruzi-specific T cells from chronic Chagas disease patients
spellingShingle Methodological approach to the ex vivo expansion and detection of T. cruzi-specific T cells from chronic Chagas disease patients
Acevedo, Gonzalo Raúl
Chagas Disease
Memory T Cells
Trypanosoma Cruzi
T Cell Library
title_short Methodological approach to the ex vivo expansion and detection of T. cruzi-specific T cells from chronic Chagas disease patients
title_full Methodological approach to the ex vivo expansion and detection of T. cruzi-specific T cells from chronic Chagas disease patients
title_fullStr Methodological approach to the ex vivo expansion and detection of T. cruzi-specific T cells from chronic Chagas disease patients
title_full_unstemmed Methodological approach to the ex vivo expansion and detection of T. cruzi-specific T cells from chronic Chagas disease patients
title_sort Methodological approach to the ex vivo expansion and detection of T. cruzi-specific T cells from chronic Chagas disease patients
dc.creator.none.fl_str_mv Acevedo, Gonzalo Raúl
Longhi, Silvia Andrea
Bunying, Alcinette
Sabri, Nazila
Atienza, Augusto
Zago, María Paola
Santos, Radleigh
Judkowski, Valeria A.
Pinilla, Clemencia
Gomez, Karina Andrea
author Acevedo, Gonzalo Raúl
author_facet Acevedo, Gonzalo Raúl
Longhi, Silvia Andrea
Bunying, Alcinette
Sabri, Nazila
Atienza, Augusto
Zago, María Paola
Santos, Radleigh
Judkowski, Valeria A.
Pinilla, Clemencia
Gomez, Karina Andrea
author_role author
author2 Longhi, Silvia Andrea
Bunying, Alcinette
Sabri, Nazila
Atienza, Augusto
Zago, María Paola
Santos, Radleigh
Judkowski, Valeria A.
Pinilla, Clemencia
Gomez, Karina Andrea
author2_role author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Chagas Disease
Memory T Cells
Trypanosoma Cruzi
T Cell Library
topic Chagas Disease
Memory T Cells
Trypanosoma Cruzi
T Cell Library
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv 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
description 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.
publishDate 2017
dc.date.none.fl_str_mv 2017-05-26
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/43074
Acevedo, Gonzalo Raúl; Longhi, Silvia Andrea; Bunying, Alcinette; Sabri, Nazila; Atienza, Augusto; et al.; Methodological approach to the ex vivo expansion and detection of T. cruzi-specific T cells from chronic Chagas disease patients; Public Library of Science; Plos One; 12; 5; 26-5-2017; 1-19; e0184467
1932-6203
CONICET Digital
CONICET
url http://hdl.handle.net/11336/43074
identifier_str_mv Acevedo, Gonzalo Raúl; Longhi, Silvia Andrea; Bunying, Alcinette; Sabri, Nazila; Atienza, Augusto; et al.; Methodological approach to the ex vivo expansion and detection of T. cruzi-specific T cells from chronic Chagas disease patients; Public Library of Science; Plos One; 12; 5; 26-5-2017; 1-19; e0184467
1932-6203
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1371/journal.pone.0178380
info:eu-repo/semantics/altIdentifier/url/http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0178380
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Public Library of Science
publisher.none.fl_str_mv Public Library of Science
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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