Search

Minor histocompatibility antigens are the main targets of donor-derived T-cells after allogeneic stem cell transplantation. Identification of these antigens and understanding their biology are a key requisite for more insight into how graft vs. leukemia effect and graft vs. host disease could be separated. We here identified four new HLA class II-restricted minor histocompatibility antigens using whole genome association scanning. For one of the new antigens, i.e., LB-PIP4K2A-1S, we measured strong T-cell recognition of the donor variant PIP4K2A-1N when pulsed as exogenous peptide, while the endogenously expressed variant in donor EBV-B cells was not recognized. We showed that lack of T-cell recognition was caused by intracellular cleavage by a protease named asparagine endopeptidase (AEP). Furthermore, microarray gene expression analysis showed that PIP4K2A and AEP are both ubiquitously expressed in a wide variety of healthy tissues, but that expression levels of AEP were lower in primary acute myeloid leukemia (AML). In line with that, we confirmed low activity of AEP in AML cells and demonstrated that HLA-DRB1 * 03:01 positive primary AML expressing LB-PIP4K2A-1S or its donor variant PIP4K2A-1N were both recognized by specific T-cells. In conclusion, LB-PIP4K2A-1S not only represents a novel minor histocompatibility antigen but also provides evidence that donor T-cells after allogeneic stem cell transplantation can target the autologous allelic variant as leukemia-associated antigen. Furthermore, it demonstrates that endopeptidases can play a role in cell type-specific intracellular processing and presentation of HLA class II-restricted antigens, which may be explored in future immunotherapy of AML., (Copyright © 2020 Kremer, Bausenwein, Lurvink, Kremer, Rutten, van Bergen, Kretschmann, van der Meijden, Honders, Mazzeo, Watts, Mackensen, Falkenburg and Griffioen.)

Patients with leukemia who receive a T cell-depleted allogeneic stem cell graft followed by postponed donor lymphocyte infusion (DLI) can experience graft-versus-leukemia (GVL) reactivity, with a lower risk of graft-versus-host disease (GVHD). Here, we have investigated the magnitude, diversity, and specificity of alloreactive CD8 T cells in patients who developed GVL reactivity after DLI in the absence or presence of GVHD. We observed a lower magnitude and diversity of CD8 T cells for minor histocompatibility antigens (MiHAs) in patients with selective GVL reactivity without GVHD. Furthermore, we demonstrated that MiHA-specific T cell clones from patients with selective GVL reactivity showed lower reactivity against nonhematopoietic cells, even when pretreated with inflammatory cytokines. Expression analysis of MiHA-encoding genes showed that similar types of antigens were recognized in both patient groups, but in patients who developed GVHD, T cell reactivity was skewed to target broadly expressed MiHAs. As an inflammatory environment can render nonhematopoietic cells susceptible to T cell recognition, prevention of such circumstances favors induction of selective GVL reactivity without development of GVHD., Competing Interests: The authors have declared that no conflict of interest exists.

Patients with malignant diseases can be effectively treated with allogeneic hematopoietic stem cell transplantation (allo-SCT). Polymorphic peptides presented in HLA molecules, the so-called minor histocompatibility antigens (MiHA), play a crucial role in antitumor immunity as targets for alloreactive donor T cells. Identification of multiple MiHAs is essential to understand and manipulate the development of clinical responses after allo-SCT. In this study, CD8+ T-cell clones were isolated from leukemia patients who entered complete remission after allo-SCT, and MiHA-specific T-cell clones were efficiently selected for analysis of recognition of a panel of EBV-transformed B cells positive for the HLA restriction elements of the selected T-cell clones. One million single nucleotide polymorphisms (SNP) were determined in the panel cell lines and investigated for matching with the T-cell recognition data by whole genome association scanning (WGAs). Significant association with 12 genomic regions was found, and detailed analysis of genes located within these genomic regions revealed SNP disparities encoding polymorphic peptides in 10 cases. Differential recognition of patient-type, but not donor-type, peptides validated the identification of these MiHAs. Using tetramers, distinct populations of MiHA-specific CD8+ T cells were detected, demonstrating that our WGAs strategy allows high-throughput discovery of relevant targets in antitumor immunity after allo-SCT., (Copyright © 2010 AACR.)

Based on clinical observations that donor T cells specific for minor histocompatibility antigens (MiHA) ubiquitously expressed on both hematopoietic and nonhematopoietic cells were detected in patients showing evident graft-versus-leukemia/lymphoma (GVL) reactivity with no or limited coinciding graft-versus-host disease (GVHD), we hypothesized that nonhematopoietic tissues may be relatively unsusceptible to the cytotoxic effect of MiHA-specific T cells under normal, noninflammatory conditions. To test this hypothesis, we investigated the reactivity of alloreactive T cells specific for ubiquitously expressed MiHA against skin-derived primary human fibroblasts. We demonstrated that this reactivity was not merely determined by their antigen-specificity, but was highly dependent on adhesion molecule expression. ICAM-1 expression on the fibroblasts upregulated under proinflammatory conditions and induced during cross-talk with the T cells was demonstrated to be a crucial factor facilitating formation of high avidity interactions with the T cells and subsequent efficient target cell destruction. Furthermore, we provide supporting evidence for the role of ICAM-1 in vivo by demonstrating that ICAM-1 expression on nonhematopoietic target cells was dependent on the presence of infiltrating activated T cells, as was illustrated by restricted ICAM-1 expression at the sites of T cell infiltration in skin biopsies of patients with acute GVHD (aGVHD), by the absence of ICAM-1 expression in the same biopsies in areas without T cell infiltration and by the absence of ICAM-1 expression in biopsies of patients without GVHD independent of the presence of infiltrating nonactivated T cells. In conclusion, under noninflammatory conditions, nonhematopoietic tissues are unsusceptible to the GVHD reactivity of alloreactive T cells due to their inability to establish high avidity interactions. [ABSTRACT FROM AUTHOR]