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• W2025459740 abstract "You have accessJournal of UrologyKidney Cancer: Basic Research (I)1 Apr 2013173 SYSTEMATIC DESIGN OF AN EFFECTIVE ANTI-CANCER VACCINE Jan Hoffmeyer, Mohammad Habbiby, Joe Tario, and Thomas Schwaab Jan HoffmeyerJan Hoffmeyer Buffalo, NY More articles by this author , Mohammad HabbibyMohammad Habbiby Buffalo, NY More articles by this author , Joe TarioJoe Tario Buffalo, NY More articles by this author , and Thomas SchwaabThomas Schwaab Buffalo, NY More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2013.02.1553AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Dendritic Cell (DC) are the most powerful antigen presenting cells. This has made them a prime target of investigation for cancer immunotherapy. However, clinical results of previous DC vaccine trials have failed for a variety of reasons. We here present a systematic approach to designing a feasible, cost-efficient and immunologically well-defined DC vaccine for urologic patients expressing the cancer/testis antigen NY-ESO-1. METHODS For all experiments, monocytes were isolated via cold agglutination or ELUTRA from blood products from healthy donors. Monocytes were cultured in GM-CSF and IL-4 in culture flasks or gas-permeable culture bags. The impact of a cell culture gell-surface (0.5% polyHEMA ) was assessed as alternative to culture flasks or gas-permeable culture bags. Maturation cocktails consisted of either TNF-α alone or TNF-α, IL-6, IL1beta and PGE-2. DC were pulsed with flu peptide or NY-ESO-1 peptide alone or with a DEC205/ NY-ESO-1 full length fusion protein. DC phenotype was analyzed using comprehensive flow cytometry and DC function was assessed using CD8+ NY-ESO-1-specific tetramers in a CD107/ IFN-ψ double stain. RESULTS DC phenotype was improved when DC were grown in culture bags or on the gel matric and matured with the maturation cocktail. Matured DC pulsed with DEC/NY-ESO-1 fusion protein induced IFN-κ production in a dose-dependent manner. Anti-DEC205 staining after loading with the fusion protein demonstrated complete saturation of the DEC205 surface marker with the fusion protein at the therapeutic dose of 50μg. Timing of loading of the fusion protein to DC was found to be critical and highly matured DC demonstrated not only highest DEC205 expression, but also highest CD8+ T cell IFN-ψ production. When compared with a number of well-described immunogenic NY-ESO-1 peptides, DCs pulsed with peptides were unable to induce CD4+ T cell responses, while DC pulsed with the fusion product elicited strong CD4+ T cell responses. In intracellular staining assays, it became evident that the DEC205/NY-ESO-1 fusion protein was taken up intra-cellularly within 15 minutes. It co-localized with HLA-Class I, but not class II molecules. CONCLUSIONS We demonstrate a systematic approach to the design of a clinical antigen-specific DC vaccine. These results are provocative and challenge current DC biology. This DC vaccine approach deserves further investigation.responses and may play a significant role for patient undergoing immunotherapy. © 2013 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 189Issue 4SApril 2013Page: e71 Advertisement Copyright & Permissions© 2013 by American Urological Association Education and Research, Inc.MetricsAuthor Information Jan Hoffmeyer Buffalo, NY More articles by this author Mohammad Habbiby Buffalo, NY More articles by this author Joe Tario Buffalo, NY More articles by this author Thomas Schwaab Buffalo, NY More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ..." @default.
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• W2025459740 title "173 SYSTEMATIC DESIGN OF AN EFFECTIVE ANTI-CANCER VACCINE" @default.
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