title: The in vitro generation of multi-tumor antigen-specific cytotoxic T cell clones: Candidates for leukemia adoptive immunotherapy following allogeneic stem cell transplantation.
abstract: Adoptive T-cell immunotherapy is a promising approach to manage and maintain relapse-free survival of leukemia patients, especially following allogeneic stem cell transplantation. Post-transplant adoptive immunotherapy using cytotoxic T lymphocytes (CTLs) of the donor origin provide graft-versus-tumor effects, with or without graft-versus-host disease. Myeloid leukemias express immunogenic leukemia associated antigens (LAAs); such as WT-1, PRAME, MAGE, h-TERT and others, most of them are able to induce specific T cell responses whenever associated with the proper co-stimulation. We investigated the ability of a LAA-expressing hybridoma cell line to induce CTL clones in PBMCs of HLA-matched healthy donors in vitro. The CTL clones were induced by repetitive co-culture with LAAs-expressing, HLA-A*0201(+) hybrid cell line, generated by fusion of leukemia blasts to human immortalized APC (EBV-sensitized B-lymphoblastoid cell line; HMy2). The induced cytotoxic T cell clones were phenotypically and functionally characterized by pentamer analysis, IFN-γ release ELISPOT and cellular cytotoxicity assays. All T cell lines showed robust peptide recognition and functional activity when sensitized with HLA-A*0201-restricted WT-1235-243, hTERT615-624 or PRAME100-108 peptides-pulsed T2 cells, in addition to partially HLA-matched leukemia blasts. This study demonstrates the feasibility of developing multi-tumor antigen-specific T cell lines in allogeneic PBMCs in vitro, using LAA-expressing tumor/HMy2 hybrid cell line model, for potential use in leukemia adoptive immunotherapy in partially matched donor-recipient setting.
authors.affiliation: Department of Medical Microbiology, College of Medicine, University of Dammam, PO BOX 2114, Dammam 31451, Saudi Arabia. Electronic address: ysmohamed@uod.edu.sa.
authors.forename: Yehia S
authors.initials: YS
authors.lastname: Mohamed
authors.affiliation: Clinical Laboratory Department, King Fahad Hospital of the University, University of Dammam, Saudi Arabia.
authors.forename: Layla A
authors.initials: LA
authors.lastname: Bashawri
authors.affiliation: Department of Genetic Research, Institute for Research and Medical Consultations, University of Dammam, PO BOX-1982, Dammam-31441, Saudi Arabia.
authors.forename: Chittibabu
authors.initials: C
authors.lastname: Vatte
authors.affiliation: Department of Biopharmaceutics & Clinical Pharmacy, Faculty of Pharmacy, University of Jordan, Amman, Jordan.
authors.forename: Eman Y
authors.initials: EY
authors.lastname: Abu-Rish
authors.affiliation: Department of Genetic Research, Institute for Research and Medical Consultations, University of Dammam, PO BOX-1982, Dammam-31441, Saudi Arabia.
authors.forename: Cyril
authors.initials: C
authors.lastname: Cyrus
authors.affiliation: Department of Infection, Immunity and Inflammation, University of Leicester, Maurice Shock Medical Sciences Building, University Road, Leicester, LE1 9HN, UK; Department of Microbiology and Immunology, Faculty of Pharmacy, Al-Azhar University, Cairo, Egypt.
authors.forename: Wafaa S
authors.initials: WS
authors.lastname: Khalaf
authors.affiliation: Department of Infection, Immunity and Inflammation, University of Leicester, Maurice Shock Medical Sciences Building, University Road, Leicester, LE1 9HN, UK.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Browning
mesh_terms: D000951:Antigens, Neoplasm / Q000276:immunology; D018920:Coculture Techniques; D003131:Combined Modality Therapy / Q000379:methods*; D058501:Enzyme-Linked Immunospot Assay; D018380:Hematopoietic Stem Cell Transplantation / Q000379:methods*; D006801:Humans; D006822:Hybrid Cells; D016219:Immunotherapy, Adoptive / Q000379:methods*; D066298:In Vitro Techniques; D007938:Leukemia / Q000276:immunology*; D016133:Polymerase Chain Reaction; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014407:Tumor Cells, Cultured
chemical_list: D000951:Antigens, Neoplasm
keywords: Adoptive T cell transfer; Hybrid cells; Leukemia immunotherapy; TAA-specific T cells; Tumor immunotherapy