Tumor-associated-antigens or osteosarcoma cell line lysates: two efficient methods for in vitro generation of CTLs with special regard to MHC-I restriction.
The expression of tumor associated antigens (TAA) in osteosarcoma cell lines allowed us to design an in vitro model for the generation of TAA-specific CTLs. Since the MHC-I-restriction of these peptides represents the major obstacle to clinical applications, we studied a second method for the generation of CTLs starting from osteosarcoma cell line lysates and PBMCs of HLA-I compatible healthy donors. TAA-specific CTLs showed high and homogeneous cytotoxic response against each peptide; high levels of IFN-γ were released by osteosarcoma cell line lysate specific-CTLs in response to the osteosarcoma cell line they were activated for. The MHC-I dependent osteosarcoma cell line lysate-specific CTLs activity was proved by the indifference against the HLA-I-negative erytroleukaemia cell line K562 and by the absence of IFN-γ production with the addition of HLA-class I blocking antibodies. These two methods may be considered the model for the autologous setting in the context of immunotherapeutic approaches for osteosarcoma patients.
Stem Cell Transplantation and Cellular Therapy Unit, Pediatric Onco-Hematology Division, Regina Margherita Children's Hospital, Turin, Italy. michela.muraro@unito.it
Michela
M
Muraro
Oana Madalina
OM
Mereuta
Francesco
F
Saglio
Francesca
F
Carraro
Cristina
C
Cravero
Franca
F
Fagioli
D019138:Antibodies, Blocking / Q000276:immunology; D000951:Antigens, Neoplasm / Q000276:immunology*; D001859:Bone Neoplasms / Q000276:immunology*; D002457:Cell Extracts / Q000276:immunology; D045744:Cell Line, Tumor; D003713:Dendritic Cells / Q000276:immunology; D015395:Histocompatibility Antigens Class I / Q000276:immunology*; D006801:Humans; D007371:Interferon-gamma / Q000276:immunology / Q000378:metabolism; D020014:K562 Cells; D008213:Lymphocyte Activation / Q000276:immunology*; D009000:Monocytes / Q000276:immunology; D012516:Osteosarcoma / Q000276:immunology*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*
D019138:Antibodies, Blocking; D000951:Antigens, Neoplasm; D002457:Cell Extracts; D015395:Histocompatibility Antigens Class I; D007371:Interferon-gamma