title: Mutated Ras-transfected, EBV-transformed lymphoblastoid cell lines as a model tumor vaccine for boosting T-cell responses against pancreatic cancer: a pilot trial.
abstract: Genetically modified lymphoblastoid cell lines (LCL) have been shown to be an attractive alternative source of antigen-presenting cells for cancer vaccination in vitro. We tested their application in patients with pancreatic cancer in a phase I clinical trial. As a model tumor antigen, we selected the point-mutated (codon 12) Ki-Ras p21 oncogene (muRas) frequently (∼85%) present in pancreatic adenocarcinoma. Autologous LCLs were established in vitro by spontaneous outgrowth from peripheral blood lymphocytes of seven pancreatic carcinoma patients and were genetically modified with an episomal Epstein-Barr virus (EBV)-based expression vector to express muRas (muRas-LCL). Weekly vaccinations with subcutaneous injection of 5×10(6) muRas-LCL were done. In six of seven patients, therapeutic vaccination elicited a T-cell response with an increase in the frequency of muRas-specific precursor cytotoxic T lymphocytes in the peripheral blood and positive delayed-type hypersensitivity reactions at the injection site. Besides local reactions and flu-like symptoms, there were no signs of toxicity and no acute EBV infection, onset of EBV-associated lymphoma, or other severe complications. A clinical response (stable disease) was observed for a short time period (2-4 months) in four of seven patients (57%), mostly in earlier tumor stages. Our results indicate that LCL presenting genetically modified antigen represent a valuable and easily available tool for in vivo autologous tumor vaccination. LCL can be transfected with any known tumor antigen and therefore should be further clinically investigated.
authors.affiliation: Department of Internal Medicine I, Jose-Carreras Centrum for Immuno- and Gene Therapy, University of Saarland Medical School, D-66421 Homburg/Saar, Germany.
authors.forename: Boris
authors.initials: B
authors.lastname: Kubuschok
authors.forename: Michael
authors.initials: M
authors.lastname: Pfreundschuh
authors.forename: Rainer
authors.initials: R
authors.lastname: Breit
authors.forename: Frank
authors.initials: F
authors.lastname: Hartmann
authors.forename: Martina
authors.initials: M
authors.lastname: Sester
authors.forename: Barbara
authors.initials: B
authors.lastname: Gärtner
authors.forename: Jochem
authors.initials: J
authors.lastname: König
authors.forename: Niels
authors.initials: N
authors.lastname: Murawski
authors.forename: Gerhard
authors.initials: G
authors.lastname: Held
authors.forename: Carsten
authors.initials: C
authors.lastname: Zwick
authors.forename: Frank
authors.initials: F
authors.lastname: Neumann
mesh_terms: D000230:Adenocarcinoma / Q000276:immunology* / Q000401:mortality / Q000628:therapy; D000368:Aged; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D019496:Cancer Vaccines / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D002461:Cell Line, Transformed; D017024:Chemotherapy, Adjuvant; D016207:Cytokines / Q000276:immunology / Q000378:metabolism; D005260:Female; D011905:Genes, ras; D004854:Herpesvirus 4, Human / Q000235:genetics / Q000276:immunology; D006801:Humans; D007279:Injections, Subcutaneous; D008297:Male; D008875:Middle Aged; D010190:Pancreatic Neoplasms / Q000276:immunology* / Q000401:mortality / Q000628:therapy*; D016577:Pancreaticoduodenectomy; D010865:Pilot Projects; D013601:T-Lymphocytes / Q000276:immunology*; D014162:Transfection; D016896:Treatment Outcome
chemical_list: D019496:Cancer Vaccines; D016207:Cytokines