An optimized method for manufacturing a clinical scale dendritic cell-based vaccine for the treatment of glioblastoma.

Immune-based treatments represent a promising new class of therapy designed to boost the immune system to specifically eradicate malignant cells. Immunotherapy may generate specific anti-tumor immune responses, and dendritic cells (DC), professional antigen-presenting cells, are widely used in exper...

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Autores principales:	Sara Nava, Marta Dossena, Simona Pogliani, Serena Pellegatta, Carlo Antozzi, Fulvio Baggi, Cinzia Gellera, Bianca Pollo, Eugenio A Parati, Gaetano Finocchiaro, Simona Frigerio
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2012
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/9a178412ea32421cb631861727bb261b
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spelling	oai:doaj.org-article:9a178412ea32421cb631861727bb261b2021-11-18T08:04:12ZAn optimized method for manufacturing a clinical scale dendritic cell-based vaccine for the treatment of glioblastoma.1932-620310.1371/journal.pone.0052301https://doaj.org/article/9a178412ea32421cb631861727bb261b2012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23284979/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Immune-based treatments represent a promising new class of therapy designed to boost the immune system to specifically eradicate malignant cells. Immunotherapy may generate specific anti-tumor immune responses, and dendritic cells (DC), professional antigen-presenting cells, are widely used in experimental cancer immunotherapy. Several reports describe methods for the generation of mature, antigen-pulsed DC for clinical use. Improved quality and standardization are desirable to obtain GMP-compliant protocols. In this study we describe the generation of DC from 31 Glioblastoma (GB) patients starting from their monocytes isolated by immunomagnetic CD14 selection using the CliniMACS® device. Upon differentiation of CD14+ with IL-4 and GM-CSF, DC were induced to maturation with TNF-α, PGE(2), IL-1β, and IL-6. Whole tumor lysate was obtained, for the first time, in a closed system using the semi-automated dissociator GentleMACS®. The yield of proteins improved by 130% compared to the manual dissociation method. Interestingly the Mean Fluorescence Intensity for CD83 increased significantly in DC pulsed with "new method" lysate compared to DC pulsed with "classical method" lysate. Our results indicate that immunomagnetic isolation of CD14(+) monocytes using the CliniMACS® device and their pulsing with whole tumor lysate proteins is a suitable method for clinical-scale generation of high quality, functional DC under GMP-grade conditions.Sara NavaMarta DossenaSimona PoglianiSerena PellegattaCarlo AntozziFulvio BaggiCinzia GelleraBianca PolloEugenio A ParatiGaetano FinocchiaroSimona FrigerioPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 12, p e52301 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Sara Nava Marta Dossena Simona Pogliani Serena Pellegatta Carlo Antozzi Fulvio Baggi Cinzia Gellera Bianca Pollo Eugenio A Parati Gaetano Finocchiaro Simona Frigerio An optimized method for manufacturing a clinical scale dendritic cell-based vaccine for the treatment of glioblastoma.
description	Immune-based treatments represent a promising new class of therapy designed to boost the immune system to specifically eradicate malignant cells. Immunotherapy may generate specific anti-tumor immune responses, and dendritic cells (DC), professional antigen-presenting cells, are widely used in experimental cancer immunotherapy. Several reports describe methods for the generation of mature, antigen-pulsed DC for clinical use. Improved quality and standardization are desirable to obtain GMP-compliant protocols. In this study we describe the generation of DC from 31 Glioblastoma (GB) patients starting from their monocytes isolated by immunomagnetic CD14 selection using the CliniMACS® device. Upon differentiation of CD14+ with IL-4 and GM-CSF, DC were induced to maturation with TNF-α, PGE(2), IL-1β, and IL-6. Whole tumor lysate was obtained, for the first time, in a closed system using the semi-automated dissociator GentleMACS®. The yield of proteins improved by 130% compared to the manual dissociation method. Interestingly the Mean Fluorescence Intensity for CD83 increased significantly in DC pulsed with "new method" lysate compared to DC pulsed with "classical method" lysate. Our results indicate that immunomagnetic isolation of CD14(+) monocytes using the CliniMACS® device and their pulsing with whole tumor lysate proteins is a suitable method for clinical-scale generation of high quality, functional DC under GMP-grade conditions.
format	article
author	Sara Nava Marta Dossena Simona Pogliani Serena Pellegatta Carlo Antozzi Fulvio Baggi Cinzia Gellera Bianca Pollo Eugenio A Parati Gaetano Finocchiaro Simona Frigerio
author_facet	Sara Nava Marta Dossena Simona Pogliani Serena Pellegatta Carlo Antozzi Fulvio Baggi Cinzia Gellera Bianca Pollo Eugenio A Parati Gaetano Finocchiaro Simona Frigerio
author_sort	Sara Nava
title	An optimized method for manufacturing a clinical scale dendritic cell-based vaccine for the treatment of glioblastoma.
title_short	An optimized method for manufacturing a clinical scale dendritic cell-based vaccine for the treatment of glioblastoma.
title_full	An optimized method for manufacturing a clinical scale dendritic cell-based vaccine for the treatment of glioblastoma.
title_fullStr	An optimized method for manufacturing a clinical scale dendritic cell-based vaccine for the treatment of glioblastoma.
title_full_unstemmed	An optimized method for manufacturing a clinical scale dendritic cell-based vaccine for the treatment of glioblastoma.
title_sort	optimized method for manufacturing a clinical scale dendritic cell-based vaccine for the treatment of glioblastoma.
publisher	Public Library of Science (PLoS)
publishDate	2012
url	https://doaj.org/article/9a178412ea32421cb631861727bb261b
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An optimized method for manufacturing a clinical scale dendritic cell-based vaccine for the treatment of glioblastoma.

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