A class II‐restricted cytotoxic T‐cell clone recognizes a human minor histocompatibility antigen with a restricted tissue distribution

Abstract

SummaryFollowing a human leucocyte antigen (HLA)-identical allogeneic stem cell transplantation (allo-SCT), minor histocompatibility antigens (mHags) play an important role in the induction of graft-versus-leukaemia (GvL) and graftversus-myeloma (GvM). Many mHags show ubiquitous tissue expression and are associated with GvL and graft-versus-host disease. Here we describe a cytotoxic CD4 + T lymphocyte line and a cytotoxic, CD4 + T cell clone (CTC), 3AB11, which recognized a tissue-restricted mHag. This CTC was isolated from a multiple myeloma patient with clinical GvM following an HLAmatched allo-SCT. CTC 3AB11 was activated in a HLA-DP*0401 restricted fashion and the antigen was expressed by 27% of HLA-DP*0401 positive Epstein-Barr virus (EBV)-transformed B-cell lines (EBV-B). Tissue distribution analysis of antigen 3AB11 showed it to be expressed by patient-derived EBV-transformed B cell lines (EBVp), the myeloma plasma cell-line UM9 and monocytes. It was weakly expressed by peripheral bloodderived phytohaemagglutinin-induced T-cell blasts and absent on CD40L stimulated peripheral B (CD40L B) cells and stromal cells. The relatively high prevalence of the HLA class II-restricted 3AB11 antigen, together with its apparent haematopoietic-restricted expression, makes it an antigen of interest for cellular immunotherapy.

Rapid Identification of Clinical Relevant Minor Histocompatibility Antigens via Genome-Wide Zygosity-Genotype Correlation Analysis

“…The HLA-DRB1*1501 restricted CD4 + T-cell clone 1GF5 was previously isolated from a multiple myeloma patient (12). It was expanded using a feeder cell cytokine mixture as previously described (13).…”

Accessory Cells of the Microenvironment Protect Multiple Myeloma from T-Cell Cytotoxicity through Cell Adhesion-Mediated Immune Resistance

“…The luciferase (Luc)-transduced human MM cell lines, U266 and UM9, were maintained in RPMI-1640 (Invitrogen), supplemented with 10% FBS (Integro BV) and antibiotics (penicillin/streptomycin; Life Technologies) as previously described (18). The authenticity was confirmed by partial HLA typing carried out maximal 6 months before the most recent experiment.…”

“…We used cloned CD4 þ and CD8 þ CTLs that are directed at different mHag expressed on multiple myeloma cells, but not on accessory cells. Because the recognition of mHags is HLA restricted, these CTLs recognize only those multiple myeloma cells expressing both the relevant antigen as well as the presenting HLA molecule (18). Accordingly, to conduct our experiments, we selected UM9 as the relevant target cell for CD4 þ CTLs and U266 for the CD8 þ CTLs (Supplementary Fig.…”

“…The HLA-DP4 restricted, minor histocompatibility antigen (mHag)-specific CD4 þ CTL clone 3AB11; the HLA-A2-restricted mHag-specific CD8 þ CTL clone HA-1; the HLA-A2-specific CD8 þ CTL clone 1E4; and the HLA-A2-restricted WT-1-specific T-cell receptor transferred polyclonal CTLs were previously described in detail with respect to their antigen-specific and HLA-restricted cytotoxic capacity against the relevant HLA-matched and antigen-positive multiple myeloma cells (18)(19)(20)(21). CTLs were expanded using a feeder cell-cytokine mixture and cryopreserved until use as described (19).…”

Identification of minor histocompatibility antigens based on the 1000 Genomes Project

“…29 In short, T cells were isolated from a multiple myeloma patient after allo-SCT. Several Tcell clones were generated of which 3AB11 displayed patient-specific cytotoxicity and was selected for additional experiments.…”