title: Adoptive transfer of chimeric antigen receptor re-directed cytolytic T lymphocyte clones in patients with neuroblastoma.
abstract: Metastatic neuroblastoma is a poor-prognosis malignancy arising during childhood that overexpresses the L1-cell adhesion molecule (CD171). We have previously described a tumor L1-cell adhesion molecule-specific, single chain antibody-derived, chimeric antigen receptor designated CE7R for re-directing the antigen-specific effector functioning of cytolytic T lymphocytes. Here, we report on the feasibility of isolating, and the safety of infusing, autologous CD8(+) cytolytic T lymphocyte clones co-expressing CE7R and the selection-suicide expression enzyme HyTK in children with recurrent/refractory neuroblastoma. The cytolytic T lymphocyte products were derived from peripheral blood mononuclear cells that were subjected to polyclonal activation, plasmid vector electrotransfer, limiting dilution hygromycin selection, and expansion to numbers sufficient for adoptive transfer. In total, 12 infusions (nine at 10(8) cells/m(2), three at 10(9) cells/m(2)) were administered to six patients. No overt toxicities to tissues known to express L1-cell adhesion molecule (e.g., central nervous system, adrenal medulla, and sympathetic ganglia) were observed. The persistence of cytolytic T lymphocyte clones in the circulation, measured by vector-specific quantitative polymerase chain reaction, was short (1-7 days) in patients with bulky disease, but significantly longer (42 days) in a patient with a limited disease burden. This first-in-humans pilot study sets the stage for clinical trials employing adoptive transfer in the context of minimal residual disease.
authors.affiliation: Department of Pediatric Hematology-Oncology, Children's Hospital and Medical Center, Seattle, Washington, USA.
authors.forename: Julie R
authors.initials: JR
authors.lastname: Park
authors.forename: David L
authors.initials: DL
authors.lastname: Digiusto
authors.forename: Marilyn
authors.initials: M
authors.lastname: Slovak
authors.forename: Christine
authors.initials: C
authors.lastname: Wright
authors.forename: Araceli
authors.initials: A
authors.lastname: Naranjo
authors.forename: Jamie
authors.initials: J
authors.lastname: Wagner
authors.forename: Hunsar B
authors.initials: HB
authors.lastname: Meechoovet
authors.forename: Cherrilyn
authors.initials: C
authors.lastname: Bautista
authors.forename: Wen-Chung
authors.initials: WC
authors.lastname: Chang
authors.forename: Julie R
authors.initials: JR
authors.lastname: Ostberg
authors.forename: Michael C
authors.initials: MC
authors.lastname: Jensen
mesh_terms: D000818:Animals; D001483:Base Sequence; D002470:Cell Survival; D002999:Clone Cells / Q000276:immunology; D017931:DNA Primers / Q000235:genetics; D005822:Genetic Vectors; D006801:Humans; D016219:Immunotherapy, Adoptive / Q000009:adverse effects / Q000379:methods*; D051379:Mice; D039842:Neural Cell Adhesion Molecule L1 / Q000235:genetics / Q000276:immunology*; D009447:Neuroblastoma / Q000276:immunology / Q000628:therapy*; D010957:Plasmids / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D012449:Safety; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D013997:Time Factors
chemical_list: D017931:DNA Primers; D039842:Neural Cell Adhesion Molecule L1; D011993:Recombinant Fusion Proteins