title: In vitro stimulation with WT1 peptide-loaded Epstein-Barr virus-positive B cells elicits high frequencies of WT1 peptide-specific T cells with in vitro and in vivo tumoricidal activity.
abstract: The Wilms tumor protein (WT1) is overexpressed in most acute and chronic leukemias. To develop a practicable, clinically applicable approach for generation of WT1-specific T cells and to comparatively evaluate the immunogenicity of WT1 in normal individuals, we sensitized T cells from 13 HLA-A0201+ and 5 HLA-A2402+ donors with autologous EBV-transformed B cells or cytokine-activated monocytes, loaded with the HLA-A0201-binding WT1 peptides (126-134)RMFPNAPYL or (187-195)SLGEQQYSV or a newly identified HLA-A2402-binding WT1 peptide (301-310)RVPGVAPTL. WT1-specific T cells were regularly generated from each donor. T cells sensitized with peptide-loaded EBV-transformed B cells generated higher numbers of WT1-specific T cells than peptide-loaded cytokine-activated monocytes. Contrary to expectations, the frequencies of WT1 peptide-specific T cells were equivalent to those generated against individual highly immunogenic HLA-A0201-binding EBV peptides. Each of these T-cell lines specifically killed WT1+ leukemias and solid tumors in an HLA-restricted manner but did not lyse autologous or HLA-matched normal CD34+ hematopoietic progenitor cells or reduce their yield of colony-forming unit-granulocyte-macrophage (CFU-GM), burst-forming unit erythroid (BFU-E), or mixed colonies (CFU-mix). Furthermore, WT1 peptide-specific T cells after adoptive transfer into nonobese diabetic-severe combined immunodeficient mice bearing subcutaneous xenografts of WT1+ and WT1- HLA-A0201+ leukemias preferentially accumulated in and induced regressions of WT1+ leukemias that expressed the restricting HLA allele. Such cells are clinically applicable and may prove useful for adoptive cell therapy of WT1+ malignant diseases in humans.
authors.affiliation: Bone Marrow Transplantation Service, Memorial Sloan-Kettering Cancer Center, New York, New York 10021, USA.
authors.forename: Ekaterina S
authors.initials: ES
authors.lastname: Doubrovina
authors.forename: Mikhail M
authors.initials: MM
authors.lastname: Doubrovin
authors.forename: Sangyull
authors.initials: S
authors.lastname: Lee
authors.forename: Jae-Hung
authors.initials: JH
authors.lastname: Shieh
authors.forename: Glen
authors.initials: G
authors.lastname: Heller
authors.forename: Eric
authors.initials: E
authors.lastname: Pamer
authors.forename: Richard J
authors.initials: RJ
authors.lastname: O'Reilly
mesh_terms: D000483:Alleles; D000818:Animals; D018952:Antigens, CD34 / Q000096:biosynthesis; D045744:Cell Line, Tumor; D002465:Cell Movement; D016207:Cytokines / Q000378:metabolism; D016162:Genes, Wilms Tumor; D004854:Herpesvirus 4, Human / Q000235:genetics*; D006801:Humans; D007091:Image Processing, Computer-Assisted; D051379:Mice; D016513:Mice, SCID; D009000:Monocytes / Q000378:metabolism; D009368:Neoplasm Transplantation; D010455:Peptides / Q000737:chemistry; D011485:Protein Binding; D012680:Sensitivity and Specificity; D013601:T-Lymphocytes / Q000276:immunology* / Q000378:metabolism; D013997:Time Factors; D025721:WT1 Proteins / Q000737:chemistry / Q000494:pharmacology*
chemical_list: D018952:Antigens, CD34; D016207:Cytokines; D010455:Peptides; D025721:WT1 Proteins