title: Ex vivo generation of human anti-pre-B leukemia-specific autologous cytolytic T cells.
abstract: In contrast to other neoplasms, antigen-specific autologous cytolytic T cells have not been detected in patients with human pre-B-cell leukemias. The absence of efficient B7 family (B7-1/CD80; B7-2/CD86) -mediated costimulation has been shown to be a major defect in tumor cells' capacity to function as antigen-presenting cells. We show here the generation of autologous anti-pre-B-cell leukemia-specific cytolytic T-cell lines from the marrows of 10 of 15 patients with pre-B-cell malignancies. T-cell costimulation via CD28 is an absolute requirement for the generation of these autologous cytolytic T cells (CTL). Although costimulation could be delivered by either bystander B7 transfectants or professional antigen-presenting cells (indirect costimulation), optimal priming and CTL expansion required that the costimulatory signal was expressed by the tumor cell (direct costimulation). These anti-pre-B-cell leukemia-specific CTL lysed both unstimulated and CD40-stimulated tumor cells from each patient studied but did not lyse either K562 or CD40-stimulated allogeneic B cells. Cytolysis was mediated by the induction of tumor cell apoptosis by CD8+ T cells via the perforin-granzyme pathway. Although we were able to generate anti-leukemia-specific CTL from the bone marrow, we were unable to generate such CTL from the peripheral blood of these patients. These studies show that antigen-specific CTL can be generated from the bone marrow of patients with pre-B-cell leukemias and these findings should facilitate the design of adoptive T-cell-mediated immunotherapy trials for the treatment of patients with B-cell precursor malignancies.
authors.affiliation: Department of Medical Oncology, Dana Farber Cancer Institute, and Brigham and Women's Hospital and Harvard Medical School, Boston, MA 02115, USA.
authors.forename: A A
authors.initials: AA
authors.lastname: Cardoso
authors.forename: M J
authors.initials: MJ
authors.lastname: Seamon
authors.forename: H M
authors.initials: HM
authors.lastname: Afonso
authors.forename: P
authors.initials: P
authors.lastname: Ghia
authors.forename: V A
authors.initials: VA
authors.lastname: Boussiotis
authors.forename: G J
authors.initials: GJ
authors.lastname: Freeman
authors.forename: J G
authors.initials: JG
authors.lastname: Gribben
authors.forename: S E
authors.initials: SE
authors.lastname: Sallan
authors.forename: L M
authors.initials: LM
authors.lastname: Nadler
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000096:biosynthesis / Q000276:immunology; D017209:Apoptosis; D018122:B7-1 Antigen / Q000096:biosynthesis / Q000276:immunology; D051940:B7-2 Antigen; D001853:Bone Marrow / Q000276:immunology* / Q000473:pathology; D002051:Burkitt Lymphoma / Q000276:immunology* / Q000473:pathology; D016466:CHO Cells; D002478:Cells, Cultured; D006224:Cricetinae; D053938:DNA Fragmentation; D003713:Dendritic Cells / Q000276:immunology; D006801:Humans; D016130:Immunophenotyping; D007371:Interferon-gamma / Q000096:biosynthesis; D015448:Leukemia, B-Cell / Q000276:immunology* / Q000473:pathology; D007959:Lymphocyte Culture Test, Mixed; D008562:Membrane Glycoproteins / Q000096:biosynthesis / Q000276:immunology; D011289:Preleukemia / Q000276:immunology* / Q000473:pathology; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012016:Reference Values; D013601:T-Lymphocytes / Q000276:immunology*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology* / Q000473:pathology; D014407:Tumor Cells, Cultured
chemical_list: D015703:Antigens, CD; D018122:B7-1 Antigen; D051940:B7-2 Antigen; C497964:CD86 protein, human; D008562:Membrane Glycoproteins; D011993:Recombinant Fusion Proteins; D007371:Interferon-gamma