title: Efficient induction of minor histocompatibility antigen HA-1-specific cytotoxic T-cells using dendritic cells retrovirally transduced with HA-1-coding cDNA.
abstract: Cytotoxic T-cells (CTLs) specific for the hematopoietic system-restricted minor histocompatibility antigen (mHag) HA-1 efficiently lyse HA-1-positive leukemic cells without affecting nonhematopoietic cells. HA-1-specific CTLs are thus potential tools for adoptive immunotherapy of relapsed leukemia after HLA-matched-HA-1-mismatched stem cell transplantation (SCT). In vitro generation of HA-1-specific CTLs from SC donors is possible using dendritic cells (DCs) pulsed with synthetic HA-1 peptide as stimulator cells. However, this approach requires at least 6 weeks of in vitro culturing under GMP (good manufacturing practice) conditions. Our data show that in vitro induction of HA-1-specific CTLs is more rapid with the use of DCs that are retrovirally transduced with the HA-1 complementary DNA. Retrovirally transduced DCs showed functional and long-term stable expression of the HA-1 CTL epitope in primary CTL cultures. In 4 SC donors, HA-1-transduced DCs induced HA-1-specific CTLs in 14 to 21 days. The in vitro-generated CTL lines contained 6% to 9% T-cells that stained brightly with tetrameric HLA-A2/HA-1 peptide complexes (HA-1(A2) tetramer) and showed significant lysis of HA-1+ leukemic cells. The CTL induction procedure using peptide-pulsed DCs was less effective and required 28 to 35 days of T-cell culture. Thus, sustained presentation of mHag HA-1 by retrovirally transduced DCs facilitates the in vitro induction of HA-1-specific CTLs.
authors.affiliation: Department of Immunohematology and Blood Transfusion, Leiden University Medical Center, The Netherlands. t.mutis@lumc.nl
authors.forename: Tuna
authors.initials: T
authors.lastname: Mutis
authors.forename: Kamran
authors.initials: K
authors.lastname: Ghoreschi
authors.forename: Ellen
authors.initials: E
authors.lastname: Schrama
authors.forename: Janine
authors.initials: J
authors.lastname: Kamp
authors.forename: Mirjam
authors.initials: M
authors.lastname: Heemskerk
authors.forename: J H Frederik
authors.initials: JH
authors.lastname: Falkenburg
authors.forename: Martina
authors.initials: M
authors.lastname: Wilke
authors.forename: Els
authors.initials: E
authors.lastname: Goulmy
mesh_terms: D017951:Antigen Presentation / Q000276:immunology; D018929:Cell Culture Techniques / Q000379:methods; D003602:Cytotoxicity, Immunologic; D018076:DNA, Complementary; D003713:Dendritic Cells / Q000166:cytology / Q000276:immunology* / Q000378:metabolism; D018984:Epitopes, T-Lymphocyte / Q000235:genetics / Q000276:immunology; D005822:Genetic Vectors; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D006801:Humans; D008213:Lymphocyte Activation / Q000276:immunology; D015778:Minor Histocompatibility Antigens / Q000235:genetics / Q000276:immunology*; D009842:Oligopeptides / Q000235:genetics / Q000276:immunology*; D012190:Retroviridae / Q000235:genetics; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014161:Transduction, Genetic / Q000379:methods
chemical_list: D018076:DNA, Complementary; D018984:Epitopes, T-Lymphocyte; C110615:HA-1 antigen; D015778:Minor Histocompatibility Antigens; D009842:Oligopeptides