high-throughput identification of potential minor histocompatibility antigens by mhc tetramer-based screening feasibility and limitations高通量鉴定潜在的次要组织相容性抗原mhc tetramer-based筛查的可行性和局限性.pdfVIP

High-Throughput Identification of Potential Minor Histocompatibility Antigens by MHC Tetramer-Based Screening: Feasibility and Limitations 1 2 2 1 1 Pleun Hombrink *, Sine R. Hadrup , Arne Bakker , Michel G. D. Kester , J. H. Frederik Falkenburg , 1 2 1 Peter A. von dem Borne , Ton N. M. Schumacher , Mirjam H. M. Heemskerk 1 Department of Hematology, Leiden University Medical Center, Leiden, The Netherlands, 2 Division of Immunology, The Netherlands Cancer Institute, Amsterdam, The Netherlands Abstract T-cell recognition of minor histocompatibility antigens (MiHA) plays an important role in the graft-versus-tumor (GVT) effect of allogeneic stem cell transplantation (allo-SCT). However, the number of MiHA identified to date remains limited, making clinical application of MiHA reactive T-cell infusion difficult. This study represents the first attempt of genome-wide prediction of MiHA, coupled to the isolation of T-cell populations that react with these antigens. In this unbiased high-throughput MiHA screen, both the possibilities and pitfalls of this approach were investigated. First, 973 polymorphic peptides expressed by hematopoietic stem cells were predicted and screened for HLA-A2 binding. Subsequently a set of 333 high affinity HLA-A2 ligands was identified and post transplantation samples from allo-SCT patients were screened for T-cell reactivity by a combination of pMHC-tetramer-based enrichment and multi-color flow cytometry. Using this approach, 71 peptide-reactive T- cell populations were generated. The isolation of a T-cell line specifically recognizing target cells expressing the MAP4K1IMA ant