title: Ex vivo generation of dendritic cells from cryopreserved, post-induction chemotherapy, mobilized leukapheresis from pediatric patients with medulloblastoma.
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.
authors: Department of Surgery, Duke University School of Medicine, Box 103035, Durham, NC, 27710, USA. smita.nair@duke.edu.|Preston Robert Tisch Brain Tumor Center, Durham, NC, USA. smita.nair@duke.edu.
authors: Smita K
authors: SK
authors: Nair
authors: Department of Pediatrics, Duke University School of Medicine, Durham, NC, 27710, USA.
authors: Timothy
authors: T
authors: Driscoll
authors: Department of Surgery, Duke University School of Medicine, Box 103035, Durham, NC, 27710, USA.
authors: David
authors: D
authors: Boczkowski
authors: Department of Surgery, Duke University School of Medicine, Box 103035, Durham, NC, 27710, USA.
authors: Robert
authors: R
authors: Schmittling
authors: Department of Surgery, Duke University School of Medicine, Box 103035, Durham, NC, 27710, USA. RReynolds@KaleidaHealth.org.|Department of Neurosurgery, University of Buffalo, Buffalo, NY, 14222, USA. RReynolds@KaleidaHealth.org.
authors: Renee
authors: R
authors: Reynolds
authors: Department of Surgery, Duke University School of Medicine, Box 103035, Durham, NC, 27710, USA. ljohnso@upenn.edu.|Abramson Family Cancer Research Institute, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, 19104, USA. ljohnso@upenn.edu.
authors: Laura A
authors: LA
authors: Johnson
authors: Department of Surgery, Duke University School of Medicine, Box 103035, Durham, NC, 27710, USA. ggrant2@stanford.edu.|Pediatric Neurosurgery, Stanford University School of Medicine, Palo Alto, CA, 94303, USA. ggrant2@stanford.edu.
authors: Gerald
authors: G
authors: Grant
authors: Department of Surgery, Duke University School of Medicine, Box 103035, Durham, NC, 27710, USA.|Department of Pediatrics, Duke University School of Medicine, Durham, NC, 27710, USA.|Preston Robert Tisch Brain Tumor Center, Durham, NC, USA.
authors: Herbert
authors: H
authors: Fuchs
authors: Department of Surgery, Duke University School of Medicine, Box 103035, Durham, NC, 27710, USA.|Department of Pathology, Duke University School of Medicine, Durham, NC, 27710, USA.|Preston Robert Tisch Brain Tumor Center, Durham, NC, USA.
authors: Darell D
authors: DD
authors: Bigner
authors: Department of Surgery, Duke University School of Medicine, Box 103035, Durham, NC, 27710, USA.|Department of Pathology, Duke University School of Medicine, Durham, NC, 27710, USA.|Preston Robert Tisch Brain Tumor Center, Durham, NC, USA.
authors: John H
authors: JH
authors: Sampson
authors: Department of Surgery, Duke University School of Medicine, Box 103035, Durham, NC, 27710, USA.|Department of Pediatrics, Duke University School of Medicine, Durham, NC, 27710, USA.|Preston Robert Tisch Brain Tumor Center, Durham, NC, USA.
authors: Sridharan
authors: S
authors: Gururangan
authors: Department of Surgery, Duke University School of Medicine, Box 103035, Durham, NC, 27710, USA. duane.mitchell@neurosurgery.ufl.edu.|Preston Robert Tisch Brain Tumor Center, Durham, NC, USA. duane.mitchell@neurosurgery.ufl.edu.|Department of Neurosurgery, Preston A. Wells, Jr. Center for Brain Tumor Therapy, University of Florida, Gainesville, FL, 32605, USA. duane.mitchell@neurosurgery.ufl.edu.
authors: Duane A
authors: DA
authors: Mitchell
mesh_terms: D015703:Antigens, CD / Q000378:metabolism; D001932:Brain Neoplasms / Q000188:drug therapy / Q000473:pathology*; D002469:Cell Separation; D002648:Child; D018920:Coculture Techniques; D003601:Cytotoxicity Tests, Immunologic; D003713:Dendritic Cells / Q000187:drug effects / Q000473:pathology / Q000502:physiology*; D005434:Flow Cytometry; D016179:Granulocyte Colony-Stimulating Factor / Q000494:pharmacology; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D060828:Induction Chemotherapy; D051034:Inhibitor of Apoptosis Proteins / Q000235:genetics / Q000378:metabolism; D007937:Leukapheresis; D008527:Medulloblastoma / Q000188:drug therapy / Q000473:pathology*; D009000:Monocytes / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D010446:Peptide Fragments / Q000235:genetics / Q000378:metabolism; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism; D000077022:Survivin; D014161:Transduction, Genetic; D014763:Viral Matrix Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D015703:Antigens, CD; C107489:BIRC5 protein, human; D051034:Inhibitor of Apoptosis Proteins; D010446:Peptide Fragments; D010750:Phosphoproteins; D000077022:Survivin; D014763:Viral Matrix Proteins; C065418:cytomegalovirus matrix protein 65kDa; C448634:influenza matrix peptide (58-66); D016179:Granulocyte Colony-Stimulating Factor; D049452:Green Fluorescent Proteins
keywords: Adoptive T cell therapy; Medulloblastoma; Pediatric brain tumors; RNA-transfected dendritic cells