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...
| Main Authors: | , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2012-01-01
|
| Series: | PLoS ONE |
| Online Access: | https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23284979/pdf/?tool=EBI |
| _version_ | 1831583571668631552 |
|---|---|
| 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 |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-12-17T20:44:35Z |
| format | Article |
| id | doaj.art-9a178412ea32421cb631861727bb261b |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-17T20:44:35Z |
| publishDate | 2012-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-9a178412ea32421cb631861727bb261b2022-12-21T21:33:13ZengPublic Library of Science (PLoS)PLoS ONE1932-62032012-01-01712e5230110.1371/journal.pone.0052301An optimized method for manufacturing a clinical scale dendritic cell-based vaccine for the treatment of glioblastoma.Sara NavaMarta DossenaSimona PoglianiSerena PellegattaCarlo AntozziFulvio BaggiCinzia GelleraBianca PolloEugenio A ParatiGaetano FinocchiaroSimona FrigerioImmune-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.https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23284979/pdf/?tool=EBI |
| spellingShingle | 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. PLoS ONE |
| title | 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_short | 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 |
| url | https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23284979/pdf/?tool=EBI |
| work_keys_str_mv | AT saranava anoptimizedmethodformanufacturingaclinicalscaledendriticcellbasedvaccineforthetreatmentofglioblastoma AT martadossena anoptimizedmethodformanufacturingaclinicalscaledendriticcellbasedvaccineforthetreatmentofglioblastoma AT simonapogliani anoptimizedmethodformanufacturingaclinicalscaledendriticcellbasedvaccineforthetreatmentofglioblastoma AT serenapellegatta anoptimizedmethodformanufacturingaclinicalscaledendriticcellbasedvaccineforthetreatmentofglioblastoma AT carloantozzi anoptimizedmethodformanufacturingaclinicalscaledendriticcellbasedvaccineforthetreatmentofglioblastoma AT fulviobaggi anoptimizedmethodformanufacturingaclinicalscaledendriticcellbasedvaccineforthetreatmentofglioblastoma AT cinziagellera anoptimizedmethodformanufacturingaclinicalscaledendriticcellbasedvaccineforthetreatmentofglioblastoma AT biancapollo anoptimizedmethodformanufacturingaclinicalscaledendriticcellbasedvaccineforthetreatmentofglioblastoma AT eugenioaparati anoptimizedmethodformanufacturingaclinicalscaledendriticcellbasedvaccineforthetreatmentofglioblastoma AT gaetanofinocchiaro anoptimizedmethodformanufacturingaclinicalscaledendriticcellbasedvaccineforthetreatmentofglioblastoma AT simonafrigerio anoptimizedmethodformanufacturingaclinicalscaledendriticcellbasedvaccineforthetreatmentofglioblastoma AT saranava optimizedmethodformanufacturingaclinicalscaledendriticcellbasedvaccineforthetreatmentofglioblastoma AT martadossena optimizedmethodformanufacturingaclinicalscaledendriticcellbasedvaccineforthetreatmentofglioblastoma AT simonapogliani optimizedmethodformanufacturingaclinicalscaledendriticcellbasedvaccineforthetreatmentofglioblastoma AT serenapellegatta optimizedmethodformanufacturingaclinicalscaledendriticcellbasedvaccineforthetreatmentofglioblastoma AT carloantozzi optimizedmethodformanufacturingaclinicalscaledendriticcellbasedvaccineforthetreatmentofglioblastoma AT fulviobaggi optimizedmethodformanufacturingaclinicalscaledendriticcellbasedvaccineforthetreatmentofglioblastoma AT cinziagellera optimizedmethodformanufacturingaclinicalscaledendriticcellbasedvaccineforthetreatmentofglioblastoma AT biancapollo optimizedmethodformanufacturingaclinicalscaledendriticcellbasedvaccineforthetreatmentofglioblastoma AT eugenioaparati optimizedmethodformanufacturingaclinicalscaledendriticcellbasedvaccineforthetreatmentofglioblastoma AT gaetanofinocchiaro optimizedmethodformanufacturingaclinicalscaledendriticcellbasedvaccineforthetreatmentofglioblastoma AT simonafrigerio optimizedmethodformanufacturingaclinicalscaledendriticcellbasedvaccineforthetreatmentofglioblastoma |