title: Identification of Biological Relevant Minor Histocompatibility Antigens within the B-lymphocyte-Derived HLA-Ligandome Using a Reverse Immunology Approach.
abstract: OBJECTIVE
  T-cell recognition of minor histocompatibility antigens (MiHA) not only plays an important role in the beneficial graft-versus-leukemia (GVL) effect of allogeneic stem cell transplantation (allo-SCT) but also mediates serious GVH complications associated with allo-SCT. Using a reverse immunology approach, we aim to develop a method enabling the identification of T-cell responses directed against predefined antigens, with the goal to select those MiHAs that can be used clinically in combination with allo-SCT.
  
  
  METHODS
  In this study, we used a recently developed MiHA selection algorithm to select candidate MiHAs within the HLA-presented ligandome of transformed B cells. From the HLA-presented ligandome that predominantly consisted of monomorphic peptides, 25 polymorphic peptides with a clinically relevant allele frequency were selected. By high-throughput screening, the availability of high-avidity T cells specific for these MiHA candidates in different healthy donors was analyzed.
  
  
  RESULTS
  With the use of MHC multimer enrichment, analyses of expanded T cells by combinatorial coding MHC multimer flow cytometry, and subsequent single-cell cloning, positive T-cell clones directed to two new MiHA: LB-CLYBL-1Y and LB-TEP1-1S could be demonstrated, indicating the immunogenicity of these two MiHAs.
  
  
  CONCLUSIONS
  The biologic relevance of MiHA LB-CLYBL-1Y was demonstrated by the detection of LB-CLYBL-1Y-specific T cells in a patient suffering from acute myeloid leukemia (AML) that experienced an anti-leukemic response after treatment with allo-SCT.
authors.affiliation: Department of Hematology, Leiden University Medical Center, Leiden, the Netherlands.
authors.forename: Pleun
authors.initials: P
authors.lastname: Hombrink
authors.affiliation: Department of Immunohematology and Blood Transfusion, Leiden University Medical Center, Leiden, the Netherlands.
authors.forename: Chopie
authors.initials: C
authors.lastname: Hassan
authors.affiliation: Department of Hematology, Leiden University Medical Center, Leiden, the Netherlands.
authors.forename: Michel G D
authors.initials: MG
authors.lastname: Kester
authors.affiliation: Department of Hematology, Leiden University Medical Center, Leiden, the Netherlands.
authors.forename: Lorenz
authors.initials: L
authors.lastname: Jahn
authors.affiliation: Department of Hematology, Leiden University Medical Center, Leiden, the Netherlands.
authors.forename: Margot J
authors.initials: MJ
authors.lastname: Pont
authors.affiliation: Department of Immunohematology and Blood Transfusion, Leiden University Medical Center, Leiden, the Netherlands.
authors.forename: Arnoud H
authors.initials: AH
authors.lastname: de Ru
authors.affiliation: Department of Hematology, Leiden University Medical Center, Leiden, the Netherlands.
authors.forename: Cornelis A M
authors.initials: CA
authors.lastname: van Bergen
authors.affiliation: Department of Hematology, Leiden University Medical Center, Leiden, the Netherlands.
authors.forename: Marieke
authors.initials: M
authors.lastname: Griffioen
authors.affiliation: Department of Hematology, Leiden University Medical Center, Leiden, the Netherlands.
authors.forename: J H Frederik
authors.initials: JH
authors.lastname: Falkenburg
authors.affiliation: Department of Immunohematology and Blood Transfusion, Leiden University Medical Center, Leiden, the Netherlands.
authors.forename: Peter A
authors.initials: PA
authors.lastname: van Veelen
authors.affiliation: Department of Hematology, Leiden University Medical Center, Leiden, the Netherlands. M.H.M.Heemskerk@lumc.nl.
authors.forename: Mirjam H M
authors.initials: MH
authors.lastname: Heemskerk
mesh_terms: D000465:Algorithms; D064591:Allografts; D001402:B-Lymphocytes / Q000276:immunology; D018984:Epitopes, T-Lymphocyte / Q000276:immunology*; D020544:Graft vs Leukemia Effect / Q000276:immunology*; D018380:Hematopoietic Stem Cell Transplantation; D006801:Humans; D015778:Minor Histocompatibility Antigens / Q000276:immunology*; D020411:Oligonucleotide Array Sequence Analysis; D013601:T-Lymphocytes / Q000276:immunology*; D053719:Tandem Mass Spectrometry
chemical_list: D018984:Epitopes, T-Lymphocyte; D015778:Minor Histocompatibility Antigens