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. |
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| Jazyk: | angličtina |
| Rok vydání: | 2007 |
| Předmět: |
chemokine receptor CCR7
cell cloning HLA antigen class 3 Time Factors HLA antigen class 2 cell migration HLA antigen class 1 B7 antigen glycoprotein gp 100 cell maturation radiation exposure Apoptosis cytotoxic T lymphocyte CD8-Positive T-Lymphocytes Monocytes CD40 antigen fluorescence activated cell sorting necrosis immunology Epitopes cell motion Cell Movement dendritic cell vaccine membrane protein CD8+ T lymphocyte CD83 antigen Melanoma time epitope Membrane Glycoproteins HLA A antigen drug effect article SILV protein human phagocytosis Cell Differentiation melan A CCR7 protein human ultrastructure Interleukin-12 unclassified drug Interleukin-10 Neoplasm Proteins cell death cell surface cytokine release dextran monocyte gamma interferon cancer vaccine coculture gp100 Melanoma Antigen melanoma cell Receptors CCR7 in vitro study HLA antigen cell kinetics dendritic cell fluorescein isothiocyanate CD86 antigen Cross-Priming MART-1 Antigen tumor protein Antigens Neoplasm Cell Line Tumor Humans controlled study human electron microscopy cross presentation human cell chemokine gamma radiation tumor cell line Dendritic Cells vaccination gamma irradiation Coculture Techniques Gamma Rays cytology antigen specificity Chemokine CCL19 interleukin 12 macrophage inflammatory protein 3beta melanocyte protein Pmel 17 antigen expression cell vacuole pathology interleukin 10 tumor antigen MLANA protein human upregulation |
| Zdroj: | J. Transl. Med. 2007;5 Biblioteca Digital (UBA-FCEN) Universidad Nacional de Buenos Aires. Facultad de Ciencias Exactas y Naturales instacron:UBA-FCEN |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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