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Ex vivo generation of dendritic cells from cryopreserved, post-induction chemotherapy, mobilized leukapheresis from pediatric patients with medulloblastoma

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Abstract

Generation of patient-derived, autologous dendritic cells (DCs) is a critical component of cancer immunotherapy with ex vivo-generated, tumor antigen-loaded DCs. An important factor in the ability to generate DCs is the potential impact of prior therapies on DC phenotype and function. We investigated the ability to generate DCs using cells harvested from pediatric patients with medulloblastoma for potential evaluation of DC-RNA based vaccination approach in this patient population. Cells harvested from medulloblastoma patient leukapheresis following induction chemotherapy and granulocyte colony stimulating factor mobilization were cryopreserved prior to use in DC generation. DCs were generated from the adherent CD14+ monocytes using standard procedures and analyzed for cell recovery, phenotype and function. To summarize, 4 out of 5 patients (80 %) had sufficient monocyte recovery to permit DC generation, and we were able to generate DCs from 3 out of these 4 patient samples (75 %). Overall, we successfully generated DCs that met phenotypic requisites for DC-based cancer therapy from 3 out of 5 (60 %) patient samples and met both phenotypic and functional requisites from 2 out of 5 (40 %) patient samples. This study highlights the potential to generate functional DCs for further clinical treatments from refractory patients that have been heavily pretreated with myelosuppressive chemotherapy. Here we demonstrate the utility of evaluating the effect of the currently employed standard-of-care therapies on the ex vivo generation of DCs for DC-based clinical studies in cancer patients.

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Acknowledgments

This work was supported by Grants from the Department of Defense Clinical Trial Award W81XWH-10-1-0089 (DAM, SG, GG), the Pediatric Brain Tumor Foundation Institute at Duke University (DDB, JHS, DAM), and Specialized Program of Research Excellence in Brain Cancer 5P50-CA108786 (DDB, JHS).

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Author notes

  1. Renee Reynolds

    Present address: Department of Neurosurgery, University of Buffalo, Buffalo, NY, 14222, USA

  2. Laura A. Johnson

    Present address: Abramson Family Cancer Research Institute, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, 19104, USA

  3. Gerald Grant

    Present address: Pediatric Neurosurgery, Stanford University School of Medicine, Palo Alto, CA, 94303, USA

  4. Duane A. Mitchell

    Present address: Department of Neurosurgery, Preston A. Wells, Jr. Center for Brain Tumor Therapy, University of Florida, Gainesville, FL, 32605, USA

Authors and Affiliations

  1. Department of Surgery, Duke University School of Medicine, Box 103035, Durham, NC, 27710, USA

    Smita K. Nair, David Boczkowski, Robert Schmittling, Renee Reynolds, Laura A. Johnson, Gerald Grant, Herbert Fuchs, Darell D. Bigner, John H. Sampson, Sridharan Gururangan & Duane A. Mitchell

  2. Department of Pediatrics, Duke University School of Medicine, Durham, NC, 27710, USA

    Timothy Driscoll, Herbert Fuchs & Sridharan Gururangan

  3. Department of Pathology, Duke University School of Medicine, Durham, NC, 27710, USA

    Darell D. Bigner & John H. Sampson

  4. Preston Robert Tisch Brain Tumor Center, Durham, NC, USA

    Smita K. Nair, Herbert Fuchs, Darell D. Bigner, John H. Sampson, Sridharan Gururangan & Duane A. Mitchell

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  1. Smita K. Nair
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Correspondence to Smita K. Nair.

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Nair, S.K., Driscoll, T., Boczkowski, D. et al. Ex vivo generation of dendritic cells from cryopreserved, post-induction chemotherapy, mobilized leukapheresis from pediatric patients with medulloblastoma. J Neurooncol 125, 65–74 (2015). https://doi.org/10.1007/s11060-015-1890-2

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