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Abstract:

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.

Registro:

Documento: Artículo
Título: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
Autor:von Euw, E.M.; Barrio, M.M.; Furman, D.; Bianchini, M.; Levy, E.M.; Yee, C.; Li, Y.; Wainstok, R.; Mordoh, J.
Filiación:Fundación Instituto Leloir, Patricias Argentinas 435, 1405 Buenos Aires, Argentina
Centro de Investigaciones Oncológicas - FUCA, Cramer 1180, 1426 Buenos Aires, Argentina
Fred Hutchinson Cancer Research Centre, Clinical Research Division, 1100 Fairview Avenue N., Seattle, WA 98109-1024, United States
Departamento Química Biológica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Buenos Aires, Argentina
Palabras clave: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
Año:2007
Volumen:5
DOI: http://dx.doi.org/10.1186/1479-5876-5-19
Título revista:Journal of Translational Medicine
Título revista abreviado:J. Transl. Med.
ISSN:14795876
CAS:chemokine receptor CCR7, 231617-75-7; dextran, 87915-38-6, 9014-78-2; fluorescein isothiocyanate, 25168-13-2, 27072-45-3, 3326-32-7; gamma interferon, 82115-62-6; interleukin 12, 138415-13-1; macrophage inflammatory protein 3beta, 181030-14-8; Antigens, Neoplasm; CCR7 protein, human; Chemokine CCL19; Epitopes; Interleukin-10, 130068-27-8; Interleukin-12, 187348-17-0; MART-1 Antigen; MLANA protein, human; Membrane Glycoproteins; Neoplasm Proteins; Receptors, CCR7; SILV protein, human; gp100 Melanoma Antigen
PDF:https://bibliotecadigital.exactas.uba.ar/download/paper/paper_14795876_v5_n_p_vonEuw.pdf
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_14795876_v5_n_p_vonEuw

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Citas:

---------- APA ----------
von Euw, E.M., Barrio, M.M., Furman, D., Bianchini, M., Levy, E.M., Yee, C., Li, Y.,..., Mordoh, J. (2007) . 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. Journal of Translational Medicine, 5.
http://dx.doi.org/10.1186/1479-5876-5-19
---------- CHICAGO ----------
von Euw, E.M., Barrio, M.M., Furman, D., Bianchini, M., Levy, E.M., Yee, C., et al. "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" . Journal of Translational Medicine 5 (2007).
http://dx.doi.org/10.1186/1479-5876-5-19
---------- MLA ----------
von Euw, E.M., Barrio, M.M., Furman, D., Bianchini, M., Levy, E.M., Yee, C., et al. "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" . Journal of Translational Medicine, vol. 5, 2007.
http://dx.doi.org/10.1186/1479-5876-5-19
---------- VANCOUVER ----------
von Euw, E.M., Barrio, M.M., Furman, D., Bianchini, M., Levy, E.M., Yee, C., et al. 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.
http://dx.doi.org/10.1186/1479-5876-5-19