title: Establishment of stable cell lines for personalized melanoma cell vaccine.
abstract: Personalized vaccine, recognized after the failure of allogenic melanoma whole cell and lysate vaccine phase III trials, involves culturing cells from a patient's own tumor within a short duration and with less passages but with optimized expression of tumor-associated antigens (TAAs). Its feasibility is established by comparing pure cell lines generated from fresh and cryopreserved tissues (n=164) of patients with lymph node (LN) and distant metastases. Stable cell lines (from 67% of specimens) are subcultured after cryopreserving them. Pure cell lines established after eliminating fibroblasts (from 96% of the cell lines) include those from LN (69%), soft tissues including cutaneous (60%), liver (64%), lung (75%), bone (80%), brain (75%), and other sites (73%). Within 3.5 months, stable cell lines (> or =50 million cells) are established from initiating the cell culture. For LN metastases, the duration differs significantly (P2<0.05) between fresh (1.4-3.4 months) and cryopreserved (2.4-4.7 months) tissues. The expression of TAAs varies as follows: Tyrosinase (81%) >Melan-A (80%) >HMB45/gp-100 (75%) >Mel-5/TRP-1 (65%) >MAGE-1 (47%) > S-100 (28%). The number of TAAs per cell line differs between early (<7) and late (>7) passages. Among late passage cell lines, lesser percentage of cell lines express three to six antigens pointing out that early passage (<7) cell lines may be needed for antigen-targeted immunotherapy. This study provides a protocol for establishing cell lines within 2-5 months for personalized vaccine therapy for nodal and organ metastatic melanoma patients.
authors.affiliation: Cell Biology Laboratory, Patty and George Hoag Cancer Center, Hoag Memorial Hospital Presbyterian, Newport Beach, California, USA. rsselvanus@yahoo.com
authors.forename: Senthamil R
authors.initials: SR
authors.lastname: Selvan
authors.forename: Denysha J
authors.initials: DJ
authors.lastname: Carbonell
authors.forename: Abner W
authors.initials: AW
authors.lastname: Fowler
authors.forename: Andrea R
authors.initials: AR
authors.lastname: Beatty
authors.forename: Mepur H
authors.initials: MH
authors.lastname: Ravindranath
authors.forename: Robert O
authors.initials: RO
authors.lastname: Dillman
mesh_terms: D000951:Antigens, Neoplasm / Q000276:immunology / Q000378:metabolism; D019496:Cancer Vaccines / Q000276:immunology*; D045744:Cell Line, Tumor; D017326:Clinical Trials, Phase III as Topic; D005260:Female; D005347:Fibroblasts / Q000276:immunology / Q000473:pathology; D006801:Humans; D007150:Immunohistochemistry; D016233:Immunotherapy, Active / Q000379:methods; D008198:Lymph Nodes / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D008297:Male; D008545:Melanoma / Q000276:immunology* / Q000378:metabolism / Q000473:pathology* / Q000628:therapy; D009367:Neoplasm Staging; D014407:Tumor Cells, Cultured
chemical_list: D000951:Antigens, Neoplasm; D019496:Cancer Vaccines