title: [Cultivation of human tumor infiltrating lymphocytes for use in adoptive immunotherapy].
abstract: We have developed a method of in vitro stimulation of TIL and PBL using immobilized anti CD3 antibody instead of autologous tumors. After this treatment, proliferation of T cells was greatly enhanced. Separated TILs were cultured by OKT3 coated 96 well culture plate for about one week with 10% human serum and 1,000U/ml of rIL-2 supplemented RPMI-1640 medium, and then transferred to plain vessels. TIL increased in number by more than 5,000-fold after 3 weeks. PBLs obtained from 10 ml of peripheral blood were cultured in OKT3-coated culture flask (75 cm2) with 10 ml of medium for about one week, and then in plain flasks and finally expanded by culture in gas-permeable culture bags. PBL proliferated by more than 1,000-fold after 2 weeks. TIL or PBL cultured by this method contained over 90% activated T cells positive for the Leu4 marker, and HLA-DR. About 60% of these were CD8-positive and about 40% were CD4-positive. Only a small number of Leu 11-positive cells were detected in every case.
authors.affiliation: National Cancer Center Hospital.
authors.forename: T
authors.initials: T
authors.lastname: Sekine
authors.forename: T
authors.initials: T
authors.lastname: Kakizoe
authors.forename: K
authors.initials: K
authors.lastname: Tobisu
authors.forename: K
authors.initials: K
authors.lastname: Takai
authors.forename: Y
authors.initials: Y
authors.lastname: Tanaka
authors.forename: M
authors.initials: M
authors.lastname: Makuuchi
authors.forename: T
authors.initials: T
authors.lastname: Takayama
mesh_terms: D000906:Antibodies / Q000276:immunology; D000945:Antigens, Differentiation, T-Lymphocyte / Q000032:analysis / Q000276:immunology; D017252:CD3 Complex; D006528:Carcinoma, Hepatocellular / Q000628:therapy; D002292:Carcinoma, Renal Cell / Q000628:therapy; D002455:Cell Division; D002478:Cells, Cultured; D003602:Cytotoxicity, Immunologic; D006801:Humans; D007116:Immunization, Passive / Q000379:methods; D007680:Kidney Neoplasms / Q000628:therapy; D008113:Liver Neoplasms / Q000556:secondary / Q000628:therapy; D008213:Lymphocyte Activation; D009369:Neoplasms / Q000276:immunology / Q000628:therapy*; D010641:Phenotype; D011948:Receptors, Antigen, T-Cell / Q000276:immunology; D012074:Remission Induction; D013601:T-Lymphocytes / Q000276:immunology / Q000473:pathology / Q000637:transplantation*; D014182:Transplantation, Autologous
chemical_list: D000906:Antibodies; D000945:Antigens, Differentiation, T-Lymphocyte; D017252:CD3 Complex; D011948:Receptors, Antigen, T-Cell