Visualization and quantification of cytotoxicity mediated by antibodies using imaging flow cytometry

Autores
Helguera, Gustavo Fernando; Rodríguez, José A.; Luria Pérez, Rosendo; Henery, Shannon; Catterton, Paul; Bregni, Carlos; George, Thaddeus C.; Martínez Maza, Otoniel; Penichet, Manuel L.
Año de publicación
2011
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Conventional approaches for the detection of antibody dependent cell-mediated cytotoxicity (ADCC) activity rely on quantification of the release of traceable compounds from target cells or flow cytometry analysis of population-wide phenomena. We report a new method for the direct imaging and quantification of ADCC of cancer cells. The proposed method using imaging flow cytometry combines the statistical power of flow cytometry with the analytical advantages of cell imaging, providing a novel and more comprehensive perspective of effector/target cell interactions during ADCC events. With this method we can quantify and show in detail the morphological changes in target and effector cells, their apoptotic index, the physical interaction between effector and target cells, and a directional transfer of cytosolic contents from effector to target cells. As a model system we used the therapeutic anti-CD20 antibody rituximab to target CFSE labeled Ramos human Burkitt's lymphoma cells, to CMTPX-labeled human monocytic U-937 effector cells. We expect that similar studies using different effector and target cell populations may contribute to the pre-clinical evaluation of therapeutic antibodies and help to identify mechanisms that could be beneficial in the immunotherapy of cancer.
Fil: Helguera, Gustavo Fernando. University of California; Estados Unidos. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Tecnología Farmacéutica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; Argentina
Fil: Rodríguez, José A.. University of California; Estados Unidos
Fil: Luria Pérez, Rosendo. University of California; Estados Unidos. Hospital infantil de México “Federico Gómez”; México
Fil: Henery, Shannon. Amnis Corporation; Estados Unidos
Fil: Catterton, Paul. Amnis Corporation; Estados Unidos
Fil: Bregni, Carlos . Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Tecnología Farmacéutica; Argentina
Fil: George, Thaddeus C.. Amnis Corporation; Estados Unidos
Fil: Martínez Maza, Otoniel. University of California; Estados Unidos
Fil: Penichet, Manuel L.. University of California; Estados Unidos
Materia
Antibody
Adcc
Cancer
Imaging flow Cytometry
Immunotherapy
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/12910
Acceder
id CONICETDig_afe74be1c5edcb710e30f1a667d20739
oai_identifier_str oai:ri.conicet.gov.ar:11336/12910
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Visualization and quantification of cytotoxicity mediated by antibodies using imaging flow cytometryHelguera, Gustavo FernandoRodríguez, José A.Luria Pérez, RosendoHenery, ShannonCatterton, PaulBregni, Carlos George, Thaddeus C.Martínez Maza, OtonielPenichet, Manuel L.AntibodyAdccCancerImaging flow CytometryImmunotherapyhttps://purl.org/becyt/ford/3.4https://purl.org/becyt/ford/3Conventional approaches for the detection of antibody dependent cell-mediated cytotoxicity (ADCC) activity rely on quantification of the release of traceable compounds from target cells or flow cytometry analysis of population-wide phenomena. We report a new method for the direct imaging and quantification of ADCC of cancer cells. The proposed method using imaging flow cytometry combines the statistical power of flow cytometry with the analytical advantages of cell imaging, providing a novel and more comprehensive perspective of effector/target cell interactions during ADCC events. With this method we can quantify and show in detail the morphological changes in target and effector cells, their apoptotic index, the physical interaction between effector and target cells, and a directional transfer of cytosolic contents from effector to target cells. As a model system we used the therapeutic anti-CD20 antibody rituximab to target CFSE labeled Ramos human Burkitt's lymphoma cells, to CMTPX-labeled human monocytic U-937 effector cells. We expect that similar studies using different effector and target cell populations may contribute to the pre-clinical evaluation of therapeutic antibodies and help to identify mechanisms that could be beneficial in the immunotherapy of cancer.Fil: Helguera, Gustavo Fernando. University of California; Estados Unidos. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Tecnología Farmacéutica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; ArgentinaFil: Rodríguez, José A.. University of California; Estados UnidosFil: Luria Pérez, Rosendo. University of California; Estados Unidos. Hospital infantil de México “Federico Gómez”; MéxicoFil: Henery, Shannon. Amnis Corporation; Estados UnidosFil: Catterton, Paul. Amnis Corporation; Estados UnidosFil: Bregni, Carlos . Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Tecnología Farmacéutica; ArgentinaFil: George, Thaddeus C.. Amnis Corporation; Estados UnidosFil: Martínez Maza, Otoniel. University of California; Estados UnidosFil: Penichet, Manuel L.. University of California; Estados UnidosElsevier Science2011-05info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/12910Helguera, Gustavo Fernando; Rodríguez, José A.; Luria Pérez, Rosendo; Henery, Shannon; Catterton, Paul; et al.; Visualization and quantification of cytotoxicity mediated by antibodies using imaging flow cytometry; Elsevier Science; Journal Of Immunological Methods; 368; 1-2; 5-2011; 54-630022-1759enginfo:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S0022175911000585info:eu-repo/semantics/altIdentifier/doi/10.1016/j.jim.2011.03.003info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-nd/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-10T12:59:43Zoai:ri.conicet.gov.ar:11336/12910instacron: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:34982025-09-10 12:59:43.941CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Visualization and quantification of cytotoxicity mediated by antibodies using imaging flow cytometry
title Visualization and quantification of cytotoxicity mediated by antibodies using imaging flow cytometry
spellingShingle Visualization and quantification of cytotoxicity mediated by antibodies using imaging flow cytometry
Helguera, Gustavo Fernando
Antibody
Adcc
Cancer
Imaging flow Cytometry
Immunotherapy
title_short Visualization and quantification of cytotoxicity mediated by antibodies using imaging flow cytometry
title_full Visualization and quantification of cytotoxicity mediated by antibodies using imaging flow cytometry
title_fullStr Visualization and quantification of cytotoxicity mediated by antibodies using imaging flow cytometry
title_full_unstemmed Visualization and quantification of cytotoxicity mediated by antibodies using imaging flow cytometry
title_sort Visualization and quantification of cytotoxicity mediated by antibodies using imaging flow cytometry
dc.creator.none.fl_str_mv Helguera, Gustavo Fernando
Rodríguez, José A.
Luria Pérez, Rosendo
Henery, Shannon
Catterton, Paul
Bregni, Carlos
George, Thaddeus C.
Martínez Maza, Otoniel
Penichet, Manuel L.
author Helguera, Gustavo Fernando
author_facet Helguera, Gustavo Fernando
Rodríguez, José A.
Luria Pérez, Rosendo
Henery, Shannon
Catterton, Paul
Bregni, Carlos
George, Thaddeus C.
Martínez Maza, Otoniel
Penichet, Manuel L.
author_role author
author2 Rodríguez, José A.
Luria Pérez, Rosendo
Henery, Shannon
Catterton, Paul
Bregni, Carlos
George, Thaddeus C.
Martínez Maza, Otoniel
Penichet, Manuel L.
author2_role author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Antibody
Adcc
Cancer
Imaging flow Cytometry
Immunotherapy
topic Antibody
Adcc
Cancer
Imaging flow Cytometry
Immunotherapy
purl_subject.fl_str_mv https://purl.org/becyt/ford/3.4
https://purl.org/becyt/ford/3
dc.description.none.fl_txt_mv Conventional approaches for the detection of antibody dependent cell-mediated cytotoxicity (ADCC) activity rely on quantification of the release of traceable compounds from target cells or flow cytometry analysis of population-wide phenomena. We report a new method for the direct imaging and quantification of ADCC of cancer cells. The proposed method using imaging flow cytometry combines the statistical power of flow cytometry with the analytical advantages of cell imaging, providing a novel and more comprehensive perspective of effector/target cell interactions during ADCC events. With this method we can quantify and show in detail the morphological changes in target and effector cells, their apoptotic index, the physical interaction between effector and target cells, and a directional transfer of cytosolic contents from effector to target cells. As a model system we used the therapeutic anti-CD20 antibody rituximab to target CFSE labeled Ramos human Burkitt's lymphoma cells, to CMTPX-labeled human monocytic U-937 effector cells. We expect that similar studies using different effector and target cell populations may contribute to the pre-clinical evaluation of therapeutic antibodies and help to identify mechanisms that could be beneficial in the immunotherapy of cancer.
Fil: Helguera, Gustavo Fernando. University of California; Estados Unidos. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Tecnología Farmacéutica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; Argentina
Fil: Rodríguez, José A.. University of California; Estados Unidos
Fil: Luria Pérez, Rosendo. University of California; Estados Unidos. Hospital infantil de México “Federico Gómez”; México
Fil: Henery, Shannon. Amnis Corporation; Estados Unidos
Fil: Catterton, Paul. Amnis Corporation; Estados Unidos
Fil: Bregni, Carlos . Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Tecnología Farmacéutica; Argentina
Fil: George, Thaddeus C.. Amnis Corporation; Estados Unidos
Fil: Martínez Maza, Otoniel. University of California; Estados Unidos
Fil: Penichet, Manuel L.. University of California; Estados Unidos
description Conventional approaches for the detection of antibody dependent cell-mediated cytotoxicity (ADCC) activity rely on quantification of the release of traceable compounds from target cells or flow cytometry analysis of population-wide phenomena. We report a new method for the direct imaging and quantification of ADCC of cancer cells. The proposed method using imaging flow cytometry combines the statistical power of flow cytometry with the analytical advantages of cell imaging, providing a novel and more comprehensive perspective of effector/target cell interactions during ADCC events. With this method we can quantify and show in detail the morphological changes in target and effector cells, their apoptotic index, the physical interaction between effector and target cells, and a directional transfer of cytosolic contents from effector to target cells. As a model system we used the therapeutic anti-CD20 antibody rituximab to target CFSE labeled Ramos human Burkitt's lymphoma cells, to CMTPX-labeled human monocytic U-937 effector cells. We expect that similar studies using different effector and target cell populations may contribute to the pre-clinical evaluation of therapeutic antibodies and help to identify mechanisms that could be beneficial in the immunotherapy of cancer.
publishDate 2011
dc.date.none.fl_str_mv 2011-05
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/11336/12910
Helguera, Gustavo Fernando; Rodríguez, José A.; Luria Pérez, Rosendo; Henery, Shannon; Catterton, Paul; et al.; Visualization and quantification of cytotoxicity mediated by antibodies using imaging flow cytometry; Elsevier Science; Journal Of Immunological Methods; 368; 1-2; 5-2011; 54-63
0022-1759
url http://hdl.handle.net/11336/12910
identifier_str_mv Helguera, Gustavo Fernando; Rodríguez, José A.; Luria Pérez, Rosendo; Henery, Shannon; Catterton, Paul; et al.; Visualization and quantification of cytotoxicity mediated by antibodies using imaging flow cytometry; Elsevier Science; Journal Of Immunological Methods; 368; 1-2; 5-2011; 54-63
0022-1759
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S0022175911000585
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.jim.2011.03.003
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Elsevier Science
publisher.none.fl_str_mv Elsevier 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
_version_ 1842979833570656256
score 12.993085

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