Safe and reproducible preparation of functional dendritic cells for immunotherapy in glioblastoma patients

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Abstract

Cell therapy based on dendritic cells (DCs) pulsed with tumor lysate is a promising approach in addition to conventional therapy for the treatment of patients with glioblastoma (GB). The success of this approach strongly depends on the ability to generate high-quality, functionally mature DCs (mDCs), with a high level of standardization and in compliance with Good Manufacturing Practices. In the cell factory of the Carlo Besta Foundation, two phase I clinical trials on immunotherapy with tumor lysateloaded DCs as treatment for GB are ongoing. From2010 to 2014, 54 patients were enrolled in the studiesand 54 batches of DCs were prepared.Weretrospectively analyzed the results of the quality control tests carried out on each produced batch, evaluating yield of mDCs and their quality in terms of microbiological safety and immunological efficacy. ThenumberofmDCsobtained allowed the treatment of all the enrolled patients. All 54 batches were sterile, conformed to acceptable endotoxin levels, and were free of Mycoplasma species and adventitious viruses. During culture, cells maintained a high percentage of viability (87%–98%), and all batches showed high viability after thawing (mean ± SD: 94.6%±2.9%). Phenotype evaluation of mDCsshowed an evident upregulation of markers typical of DC maturation; mixed lymphocyte reaction tests for the functional evaluation of DCs demonstrated that all batches were able to induce lymphocyte responses. These results demonstrated that our protocol for DC preparation is highly reproducible and permits generation of large numbers of safe and functional DCs for in vivo use in immunotherapy approaches.

Original languageEnglish
Pages (from-to)1164-1172
Number of pages9
JournalStem cells translational medicine
Volume4
Issue number10
DOIs
Publication statusPublished - Oct 1 2015

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Glioblastoma
Immunotherapy
Dendritic Cells
Clinical Trials, Phase I
Mixed Lymphocyte Culture Test
Mycoplasma
Therapeutics
Cell- and Tissue-Based Therapy
Endotoxins
Quality Control
Neoplasms
Up-Regulation
Lymphocytes
Viruses
Phenotype
Safety

Keywords

  • Dendritic cells
  • Glioblastoma
  • Good manufacturing practices
  • Immunotherapy
  • Quality controls

ASJC Scopus subject areas

  • Cell Biology
  • Developmental Biology

Cite this

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title = "Safe and reproducible preparation of functional dendritic cells for immunotherapy in glioblastoma patients",
abstract = "Cell therapy based on dendritic cells (DCs) pulsed with tumor lysate is a promising approach in addition to conventional therapy for the treatment of patients with glioblastoma (GB). The success of this approach strongly depends on the ability to generate high-quality, functionally mature DCs (mDCs), with a high level of standardization and in compliance with Good Manufacturing Practices. In the cell factory of the Carlo Besta Foundation, two phase I clinical trials on immunotherapy with tumor lysateloaded DCs as treatment for GB are ongoing. From2010 to 2014, 54 patients were enrolled in the studiesand 54 batches of DCs were prepared.Weretrospectively analyzed the results of the quality control tests carried out on each produced batch, evaluating yield of mDCs and their quality in terms of microbiological safety and immunological efficacy. ThenumberofmDCsobtained allowed the treatment of all the enrolled patients. All 54 batches were sterile, conformed to acceptable endotoxin levels, and were free of Mycoplasma species and adventitious viruses. During culture, cells maintained a high percentage of viability (87{\%}–98{\%}), and all batches showed high viability after thawing (mean ± SD: 94.6{\%}±2.9{\%}). Phenotype evaluation of mDCsshowed an evident upregulation of markers typical of DC maturation; mixed lymphocyte reaction tests for the functional evaluation of DCs demonstrated that all batches were able to induce lymphocyte responses. These results demonstrated that our protocol for DC preparation is highly reproducible and permits generation of large numbers of safe and functional DCs for in vivo use in immunotherapy approaches.",
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author = "Sara Nava and Daniela Lisini and Simona Pogliani and Marta Dossena and Anna Bersano and Serena Pellegatta and Eugenio Parati and Gaetano Finocchiaro and Simona Frigerio",
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AU - Finocchiaro, Gaetano

AU - Frigerio, Simona

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