Monocyte-derived dendritic cells loaded with a mixture of apoptotic/ necrotic melanoma cells efficiently cross-present gp100 and MART-1 antigens to specific CD8+ T lymphocytes

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Background: In the present study, we demonstrate, in rigorous fashion, that human monocyte-derived immature dendritic cells (DCs) can efficiently cross-present tumor-associated antigens when co-cultured with a mixture of human melanoma cells rendered apoptotic/necrotic by γ irradiation (Apo-Nec cell...

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Autores principales:	von Euw, E.M., Barrio, M.M., Furman, D., Bianchini, M., Levy, E.M., Yee, C., Li, Y., Wainstok, R., Mordoh, J.
Formato:	Artículo publishedVersion
Lenguaje:	Inglés
Publicado:	2007
Materias:	B7 antigen cancer vaccine CD40 antigen CD83 antigen CD86 antigen chemokine chemokine receptor CCR7 dendritic cell vaccine dextran fluorescein isothiocyanate gamma interferon glycoprotein gp 100 HLA A antigen HLA antigen HLA antigen class 1 HLA antigen class 2 HLA antigen class 3 interleukin 10 interleukin 12 macrophage inflammatory protein 3beta melan A tumor antigen unclassified drug CCR7 protein, human epitope melanocyte protein Pmel 17 membrane protein MLANA protein, human SILV protein, human tumor protein antigen expression antigen specificity apoptosis article CD8+ T lymphocyte cell cloning cell death cell kinetics cell maturation cell migration cell surface cell vacuole coculture controlled study cross presentation cytokine release cytotoxic T lymphocyte dendritic cell electron microscopy fluorescence activated cell sorting gamma irradiation human human cell in vitro study melanoma cell monocyte phagocytosis upregulation vaccination cell differentiation cell motion cytology drug effect gamma radiation immunology melanoma necrosis pathology radiation exposure time tumor cell line ultrastructure Antigens, Neoplasm Apoptosis CD8-Positive T-Lymphocytes Cell Differentiation Cell Line, Tumor Cell Movement Chemokine CCL19 Coculture Techniques Cross-Priming Dendritic Cells Epitopes Gamma Rays gp100 Melanoma Antigen Humans Interleukin-10 Interleukin-12 MART-1 Antigen Melanoma Membrane Glycoproteins Monocytes Necrosis Neoplasm Proteins Phagocytosis Receptors, CCR7 Time Factors
Acceso en línea:	http://hdl.handle.net/20.500.12110/paper_14795876_v5_n_p_vonEuw
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

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institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
language	Inglés
orig_language_str_mv	eng
topic	B7 antigen cancer vaccine CD40 antigen CD83 antigen CD86 antigen chemokine chemokine receptor CCR7 dendritic cell vaccine dextran fluorescein isothiocyanate gamma interferon glycoprotein gp 100 HLA A antigen HLA antigen HLA antigen class 1 HLA antigen class 2 HLA antigen class 3 interleukin 10 interleukin 12 macrophage inflammatory protein 3beta melan A tumor antigen unclassified drug CCR7 protein, human chemokine receptor CCR7 epitope interleukin 10 interleukin 12 macrophage inflammatory protein 3beta melan A melanocyte protein Pmel 17 membrane protein MLANA protein, human SILV protein, human tumor antigen tumor protein antigen expression antigen specificity apoptosis article CD8+ T lymphocyte cell cloning cell death cell kinetics cell maturation cell migration cell surface cell vacuole coculture controlled study cross presentation cytokine release cytotoxic T lymphocyte dendritic cell electron microscopy fluorescence activated cell sorting gamma irradiation human human cell in vitro study melanoma cell monocyte phagocytosis upregulation vaccination apoptosis cell differentiation cell motion cross presentation cytology drug effect gamma radiation immunology melanoma monocyte necrosis pathology radiation exposure time tumor cell line ultrastructure Antigens, Neoplasm Apoptosis CD8-Positive T-Lymphocytes Cell Differentiation Cell Line, Tumor Cell Movement Chemokine CCL19 Coculture Techniques Cross-Priming Dendritic Cells Epitopes Gamma Rays gp100 Melanoma Antigen Humans Interleukin-10 Interleukin-12 MART-1 Antigen Melanoma Membrane Glycoproteins Monocytes Necrosis Neoplasm Proteins Phagocytosis Receptors, CCR7 Time Factors
spellingShingle	B7 antigen cancer vaccine CD40 antigen CD83 antigen CD86 antigen chemokine chemokine receptor CCR7 dendritic cell vaccine dextran fluorescein isothiocyanate gamma interferon glycoprotein gp 100 HLA A antigen HLA antigen HLA antigen class 1 HLA antigen class 2 HLA antigen class 3 interleukin 10 interleukin 12 macrophage inflammatory protein 3beta melan A tumor antigen unclassified drug CCR7 protein, human chemokine receptor CCR7 epitope interleukin 10 interleukin 12 macrophage inflammatory protein 3beta melan A melanocyte protein Pmel 17 membrane protein MLANA protein, human SILV protein, human tumor antigen tumor protein antigen expression antigen specificity apoptosis article CD8+ T lymphocyte cell cloning cell death cell kinetics cell maturation cell migration cell surface cell vacuole coculture controlled study cross presentation cytokine release cytotoxic T lymphocyte dendritic cell electron microscopy fluorescence activated cell sorting gamma irradiation human human cell in vitro study melanoma cell monocyte phagocytosis upregulation vaccination apoptosis cell differentiation cell motion cross presentation cytology drug effect gamma radiation immunology melanoma monocyte necrosis pathology radiation exposure time tumor cell line ultrastructure Antigens, Neoplasm Apoptosis CD8-Positive T-Lymphocytes Cell Differentiation Cell Line, Tumor Cell Movement Chemokine CCL19 Coculture Techniques Cross-Priming Dendritic Cells Epitopes Gamma Rays gp100 Melanoma Antigen Humans Interleukin-10 Interleukin-12 MART-1 Antigen Melanoma Membrane Glycoproteins Monocytes Necrosis Neoplasm Proteins Phagocytosis Receptors, CCR7 Time Factors von Euw, E.M. Barrio, M.M. Furman, D. Bianchini, M. Levy, E.M. Yee, C. Li, Y. Wainstok, R. Mordoh, J. Monocyte-derived dendritic cells loaded with a mixture of apoptotic/ necrotic melanoma cells efficiently cross-present gp100 and MART-1 antigens to specific CD8+ T lymphocytes
topic_facet	B7 antigen cancer vaccine CD40 antigen CD83 antigen CD86 antigen chemokine chemokine receptor CCR7 dendritic cell vaccine dextran fluorescein isothiocyanate gamma interferon glycoprotein gp 100 HLA A antigen HLA antigen HLA antigen class 1 HLA antigen class 2 HLA antigen class 3 interleukin 10 interleukin 12 macrophage inflammatory protein 3beta melan A tumor antigen unclassified drug CCR7 protein, human chemokine receptor CCR7 epitope interleukin 10 interleukin 12 macrophage inflammatory protein 3beta melan A melanocyte protein Pmel 17 membrane protein MLANA protein, human SILV protein, human tumor antigen tumor protein antigen expression antigen specificity apoptosis article CD8+ T lymphocyte cell cloning cell death cell kinetics cell maturation cell migration cell surface cell vacuole coculture controlled study cross presentation cytokine release cytotoxic T lymphocyte dendritic cell electron microscopy fluorescence activated cell sorting gamma irradiation human human cell in vitro study melanoma cell monocyte phagocytosis upregulation vaccination apoptosis cell differentiation cell motion cross presentation cytology drug effect gamma radiation immunology melanoma monocyte necrosis pathology radiation exposure time tumor cell line ultrastructure Antigens, Neoplasm Apoptosis CD8-Positive T-Lymphocytes Cell Differentiation Cell Line, Tumor Cell Movement Chemokine CCL19 Coculture Techniques Cross-Priming Dendritic Cells Epitopes Gamma Rays gp100 Melanoma Antigen Humans Interleukin-10 Interleukin-12 MART-1 Antigen Melanoma Membrane Glycoproteins Monocytes Necrosis Neoplasm Proteins Phagocytosis Receptors, CCR7 Time Factors
description	Background: In the present study, we demonstrate, in rigorous fashion, that human monocyte-derived immature dendritic cells (DCs) can efficiently cross-present tumor-associated antigens when co-cultured with a mixture of human melanoma cells rendered apoptotic/necrotic by γ irradiation (Apo-Nec cells). Methods: We evaluated the phagocytosis of Apo-Nec cells by FACS after PKH26 and PKH67 staining of DCs and Apo-Nec cells at different times of coculture. The kinetics of the process was also followed by electron microscopy. DCs maturation was also studied monitoring the expression of specific markers, migration towards specific chemokines and the ability to cross-present in vitro the native melanoma-associated Ags MelanA/MART-1 and gp100. Results: Apo-Nec cells were efficiently phagocytosed by immature DCs (iDC) (55 ± 10.5%) at 12 hs of coculture. By 12-24 hs we observed digested Apo-Nec cells inside DCs and large empty vacuoles as part of the cellular processing. Loading with Apo-Nec cells induced DCs maturation to levels achieved using LPS treatment, as measured by: i) the decrease in FITC - Dextran uptake (iDC: 81 ± 5%; DC/Apo-Nec 33 ± 12%); ii) the cell surface up-regulation of CD80, CD86, CD83, CCR7, CD40, HLA-I and HLA-II and iii) an increased in vitro migration towards MIP-3β. DC/Apo-Nec isolated from HLA-A*0201 donors were able to induce >600 pg/ml IFN-γ secretion of CTL clones specific for MelanA/MART-1 and gp100 Ags after 6 hs and up to 48 hs of coculture, demonstrating efficient cross-presentation of the native Ags. Intracellular IL-12 was detected in DC/Apo-Nec 24 hs post-coculture while IL-10 did not change. Conclusion: We conclude thatthe use of a mixture of four apoptotic/ necrotic melanoma cell lines is a suitable source of native melanoma Ags that provides maturation signals for DCs, increases migration to MIP-3β and allows Ag cross-presentation. This strategy could be exploited for vaccination of melanoma patients. © 2007 von Euw et al; licensee BioMed Central Ltd.
format	Artículo Artículo publishedVersion
author	von Euw, E.M. Barrio, M.M. Furman, D. Bianchini, M. Levy, E.M. Yee, C. Li, Y. Wainstok, R. Mordoh, J.
author_facet	von Euw, E.M. Barrio, M.M. Furman, D. Bianchini, M. Levy, E.M. Yee, C. Li, Y. Wainstok, R. Mordoh, J.
author_sort	von Euw, E.M.
title	Monocyte-derived dendritic cells loaded with a mixture of apoptotic/ necrotic melanoma cells efficiently cross-present gp100 and MART-1 antigens to specific CD8+ T lymphocytes
title_short	Monocyte-derived dendritic cells loaded with a mixture of apoptotic/ necrotic melanoma cells efficiently cross-present gp100 and MART-1 antigens to specific CD8+ T lymphocytes
title_full	Monocyte-derived dendritic cells loaded with a mixture of apoptotic/ necrotic melanoma cells efficiently cross-present gp100 and MART-1 antigens to specific CD8+ T lymphocytes
title_fullStr	Monocyte-derived dendritic cells loaded with a mixture of apoptotic/ necrotic melanoma cells efficiently cross-present gp100 and MART-1 antigens to specific CD8+ T lymphocytes
title_full_unstemmed	Monocyte-derived dendritic cells loaded with a mixture of apoptotic/ necrotic melanoma cells efficiently cross-present gp100 and MART-1 antigens to specific CD8+ T lymphocytes
title_sort	monocyte-derived dendritic cells loaded with a mixture of apoptotic/ necrotic melanoma cells efficiently cross-present gp100 and mart-1 antigens to specific cd8+ t lymphocytes
publishDate	2007
url	http://hdl.handle.net/20.500.12110/paper_14795876_v5_n_p_vonEuw
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spelling	paperaa:paper_14795876_v5_n_p_vonEuw2023-06-12T16:50:29Z Monocyte-derived dendritic cells loaded with a mixture of apoptotic/ necrotic melanoma cells efficiently cross-present gp100 and MART-1 antigens to specific CD8+ T lymphocytes J. Transl. Med. 2007;5 von Euw, E.M. Barrio, M.M. Furman, D. Bianchini, M. Levy, E.M. Yee, C. Li, Y. Wainstok, R. Mordoh, J. B7 antigen cancer vaccine CD40 antigen CD83 antigen CD86 antigen chemokine chemokine receptor CCR7 dendritic cell vaccine dextran fluorescein isothiocyanate gamma interferon glycoprotein gp 100 HLA A antigen HLA antigen HLA antigen class 1 HLA antigen class 2 HLA antigen class 3 interleukin 10 interleukin 12 macrophage inflammatory protein 3beta melan A tumor antigen unclassified drug CCR7 protein, human chemokine receptor CCR7 epitope interleukin 10 interleukin 12 macrophage inflammatory protein 3beta melan A melanocyte protein Pmel 17 membrane protein MLANA protein, human SILV protein, human tumor antigen tumor protein antigen expression antigen specificity apoptosis article CD8+ T lymphocyte cell cloning cell death cell kinetics cell maturation cell migration cell surface cell vacuole coculture controlled study cross presentation cytokine release cytotoxic T lymphocyte dendritic cell electron microscopy fluorescence activated cell sorting gamma irradiation human human cell in vitro study melanoma cell monocyte phagocytosis upregulation vaccination apoptosis cell differentiation cell motion cross presentation cytology drug effect gamma radiation immunology melanoma monocyte necrosis pathology radiation exposure time tumor cell line ultrastructure Antigens, Neoplasm Apoptosis CD8-Positive T-Lymphocytes Cell Differentiation Cell Line, Tumor Cell Movement Chemokine CCL19 Coculture Techniques Cross-Priming Dendritic Cells Epitopes Gamma Rays gp100 Melanoma Antigen Humans Interleukin-10 Interleukin-12 MART-1 Antigen Melanoma Membrane Glycoproteins Monocytes Necrosis Neoplasm Proteins Phagocytosis Receptors, CCR7 Time Factors Background: In the present study, we demonstrate, in rigorous fashion, that human monocyte-derived immature dendritic cells (DCs) can efficiently cross-present tumor-associated antigens when co-cultured with a mixture of human melanoma cells rendered apoptotic/necrotic by γ irradiation (Apo-Nec cells). Methods: We evaluated the phagocytosis of Apo-Nec cells by FACS after PKH26 and PKH67 staining of DCs and Apo-Nec cells at different times of coculture. The kinetics of the process was also followed by electron microscopy. DCs maturation was also studied monitoring the expression of specific markers, migration towards specific chemokines and the ability to cross-present in vitro the native melanoma-associated Ags MelanA/MART-1 and gp100. Results: Apo-Nec cells were efficiently phagocytosed by immature DCs (iDC) (55 ± 10.5%) at 12 hs of coculture. By 12-24 hs we observed digested Apo-Nec cells inside DCs and large empty vacuoles as part of the cellular processing. Loading with Apo-Nec cells induced DCs maturation to levels achieved using LPS treatment, as measured by: i) the decrease in FITC - Dextran uptake (iDC: 81 ± 5%; DC/Apo-Nec 33 ± 12%); ii) the cell surface up-regulation of CD80, CD86, CD83, CCR7, CD40, HLA-I and HLA-II and iii) an increased in vitro migration towards MIP-3β. DC/Apo-Nec isolated from HLA-A*0201 donors were able to induce >600 pg/ml IFN-γ secretion of CTL clones specific for MelanA/MART-1 and gp100 Ags after 6 hs and up to 48 hs of coculture, demonstrating efficient cross-presentation of the native Ags. Intracellular IL-12 was detected in DC/Apo-Nec 24 hs post-coculture while IL-10 did not change. Conclusion: We conclude thatthe use of a mixture of four apoptotic/ necrotic melanoma cell lines is a suitable source of native melanoma Ags that provides maturation signals for DCs, increases migration to MIP-3β and allows Ag cross-presentation. This strategy could be exploited for vaccination of melanoma patients. © 2007 von Euw et al; licensee BioMed Central Ltd. Fil:von Euw, E.M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Barrio, M.M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Levy, E.M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 2007 info:eu-repo/semantics/article info:ar-repo/semantics/artículo info:eu-repo/semantics/publishedVersion application/pdf eng info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_14795876_v5_n_p_vonEuw

Monocyte-derived dendritic cells loaded with a mixture of apoptotic/ necrotic melanoma cells efficiently cross-present gp100 and MART-1 antigens to specific CD8+ T lymphocytes

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