title: Selective depletion of alloreactive T lymphocytes using patient-derived nonhematopoietic stimulator cells in allograft engineering.
abstract: BACKGROUND
  Selective depletion of alloreactive T cells in vitro results in efficient graft-versus-host disease prophylaxis in allogeneic hematopoietic stem-cell transplantation, but it is accompanied by increased recurrence of leukemia. To spare donor T-cell-mediated graft-versus-leukemia immunity against hematopoiesis-restricted minor histocompatibility (minor-H) antigens, we explored the use of patient-derived nonhematopoietic antigen-presenting cells (APC) as allogeneic stimulators for selective allodepletion in leukemia-reactive donor T-cell lines.
  
  
  METHODS
  Primary keratinocytes, dermal fibroblasts, and bone marrow fibroblasts were generated from skin biopsies and diagnostic bone marrow aspirates of acute myeloid leukemia patients in vitro. Cell cultures were analyzed for expansion, phenotype, and immunostimulatory capacity in comparison with CD40-activated B cells as professional APC. In addition, nonhematopoietic APCs were used for selective allodepletion in vitro.
  
  
  RESULTS
  Patient-derived fibroblasts could be reliably expanded to large cell numbers, whereas keratinocytes had limited growth potential. Interferon-gamma-pretreated fibroblasts showed increased expression of human leukocyte antigen (HLA)-class I and II molecules, CD40, and CD54. Fibroblasts and CD40-activated B cells comparably stimulated HLA-A*0301-specific CD8 T cells after transient expression of HLA-A*0301 as a model alloantigen. Finally, fibroblasts could be effectively applied to selectively deplete alloreactivity within leukemia-reactive donor CD8 T-cell lines by targeting the activation-induced antigen CD137.
  
  
  CONCLUSIONS
  Primary fibroblasts can be efficiently used as allogeneic nonhematopoietic APC for selective depletion of donor T cells reactive to HLA and ubiquitously expressed minor-H antigen disparities in leukemia-stimulated CD8 T-cell lines. Therefore, harnessing alloreactivity to hematopoietic minor-H antigens in addition to leukemia-associated antigens might increase graft-versus-leukemia immunity of donor lymphocyte grafts in allogeneic hematopoietic stem-cell transplantation.
authors.affiliation: Department of Medicine III - Hematology and Oncology, Johannes Gutenberg-University Medical School, Mainz, Germany.
authors.forename: Marion
authors.initials: M
authors.lastname: Nonn
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Herr
authors.forename: Shamsul
authors.initials: S
authors.lastname: Khan
authors.forename: Mariya
authors.initials: M
authors.lastname: Todorova
authors.forename: Irina
authors.initials: I
authors.lastname: Link
authors.forename: Jochen
authors.initials: J
authors.lastname: Thies
authors.forename: Eva
authors.initials: E
authors.lastname: Distler
authors.forename: Marcus
authors.initials: M
authors.lastname: Kaltwasser
authors.forename: Julia
authors.initials: J
authors.lastname: Hoffmann
authors.forename: Christoph
authors.initials: C
authors.lastname: Huber
authors.forename: Udo F
authors.initials: UF
authors.lastname: Hartwig
mesh_terms: D000328:Adult; D015703:Antigens, CD / Q000276:immunology; D001402:B-Lymphocytes / Q000276:immunology; D020405:Dermis / Q000166:cytology / Q000276:immunology; D000078404:Epidermal Cells; D004817:Epidermis / Q000276:immunology; D005347:Fibroblasts / Q000187:drug effects / Q000276:immunology; D006086:Graft vs Host Disease / Q000276:immunology; D006681:HLA-D Antigens / Q000276:immunology; D018380:Hematopoietic Stem Cell Transplantation / Q000379:methods; D015395:Histocompatibility Antigens Class I / Q000276:immunology; D006801:Humans; D007371:Interferon-gamma / Q000276:immunology; D015603:Keratinocytes / Q000276:immunology; D008212:Lymphocyte Depletion / Q000379:methods*; D012867:Skin / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology*; D023822:Tissue Engineering / Q000379:methods; D014184:Transplantation, Homologous / Q000276:immunology*; D053261:Tumor Necrosis Factor Receptor Superfamily, Member 9 / Q000276:immunology
chemical_list: D015703:Antigens, CD; D006681:HLA-D Antigens; D015395:Histocompatibility Antigens Class I; D053261:Tumor Necrosis Factor Receptor Superfamily, Member 9; D007371:Interferon-gamma