title: High-fold expansion of human cytotoxic T-lymphocytes specific for autologous melanoma cells for use in immunotherapy.
abstract: We set up a culture protocol that consistently allows high-fold expansion of tumor-specific T-lymphocytes from most melanoma-invaded biopsies with low doses of recombinant interleukin-2 (rIL-2). Between 2-60 x 10(6) T-lymphocytes could be obtained and cryopreserved from 12 out of 13 patients, by culturing only 50 mm3 tumor tissue with rIL-2. Thawed lymphocytes from 11 of these patients could then be expanded by a median factor of 32,800 by culturing them successively in microplates on irradiated feeder cells with rIL-2 for approximately 2 weeks and then in culture bags or flasks with only rIL-2 for 1-2 additional weeks. Dead feeder cells disappeared during the last phase of the lymphocyte culture with rIL-2. Interestingly, each time they were expanded under these conditions, tumor-infiltrating lymphocytes (TIL) or lymph-node lymphocytes developed a lytic activity apparently restricted to the autologous melanoma line. Tumor-specific lysis, which was maximum at around the end of T-lymphocyte expansion, ranged between 31-63% lysis at an effector:target (E:T) ratio of 20:1. This culture method would thus appear to be suitable for reliable production of over 10(10) T-lymphocytes with good tumor-specific lytic activity from most melanoma-invaded biopsy. It should permit analysis of the immunotherapeutic potential of these populations reinjected into cancer patients.
authors.affiliation: Unité 211 INSERM, Faculté des Sciences, Nantes, France.
authors.forename: F
authors.initials: F
authors.lastname: Jotereau
authors.forename: M C
authors.initials: MC
authors.lastname: Pandolfino
authors.forename: D
authors.initials: D
authors.lastname: Boudart
authors.forename: E
authors.initials: E
authors.lastname: Diez
authors.forename: B
authors.initials: B
authors.lastname: Dreno
authors.forename: J Y
authors.initials: JY
authors.lastname: Douillard
authors.forename: J Y
authors.initials: JY
authors.lastname: Muller
authors.forename: B
authors.initials: B
authors.lastname: LeMevel
mesh_terms: D002455:Cell Division; D003602:Cytotoxicity, Immunologic; D006801:Humans; D016219:Immunotherapy, Adoptive; D016246:Lymphocytes, Tumor-Infiltrating / Q000276:immunology / Q000473:pathology; D008545:Melanoma / Q000276:immunology / Q000473:pathology / Q000628:therapy*; D010641:Phenotype; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology* / Q000473:pathology; D014407:Tumor Cells, Cultured / Q000276:immunology