Genetic engineering of virus-specific T cells with T-cell receptors recognizing minor histocompatibility antigens for clinical application

Abstract

BackgroundDonor lymphocyte infusion is an effective form of adoptive immunotherapy for hematologic malignancies after allogeneic stem cell transplantation. Graft-versus-host disease, however, often develops due to recognition of ubiquitously-expressed minor histocompatibility antigens. Transfer of T-cell receptors recognizing hematopoiesis-restricted minor histocompatibility antigens to virus-specific T cells may be a powerful anti-tumor therapy with a low risk of graft-versus-host disease. The purpose of this study was to develop an optimal T-cell receptors-encoding multi-cistronic retroviral vector and an efficient method for generating T-cell receptors-engineered virus-specific T cells.

Identification of 4 new HLA-DR–restricted minor histocompatibility antigens as hematopoietic targets in antitumor immunity

“…Viral supernatants were used for transduction of EBV-LCLs on plates coated with recombinant human fibronectin CH 296 (Takara Shuzo) as previously described. 37 All EBV-LCLs showed retroviral transduction efficiencies of 3% to 7% based on staining with PE-conjugated anti-⌬NGF-R (BD Biosciences).…”

“…Polymerase chain reaction (PCR) products were cloned into retroviral vector MP71, which contains the marker nerve growth factor receptor (⌬NGF-R). 37 PCR products were verified by sequencing. Wild-type nx A packaging cells were transfected with these vectors as previously described, 37 with the exception that the Fugene HD transfection kit (Roche Diagnostics) was used.…”

“…37 PCR products were verified by sequencing. Wild-type nx A packaging cells were transfected with these vectors as previously described, 37 with the exception that the Fugene HD transfection kit (Roche Diagnostics) was used. Viral supernatants were used for transduction of EBV-LCLs on plates coated with recombinant human fibronectin CH 296 (Takara Shuzo) as previously described.…”

“…A 15-mer peptide comprising amino acids shared by the 2 19-mer peptides also stimulated IFN-␥ release, whereas a 15-mer peptide containing the donor variant H at position 39 was not recognized ( Figure 3A). Serial truncations of the 15-mer peptide at the N and C termini led to the identification of a minimal 12-mer YFRLGVSDPIRG peptide (amino acids [29][30][31][32][33][34][35][36][37][38][39][40], designated LB-MR1-1R ( Figure 3A). …”

Human allo-reactive CD4+ T cells as strong mediators of anti-tumor immunity in NOD/scid mice engrafted with human acute lymphoblastic leukemia

“…Donor EBV-LCL and HLA-class II negative Hela cells were retrovirally transduced with the mismatched HLA-class II alleles of the patients as described previously. [26][27][28] Hela cells retrovirally transduced with the invariant chain (Ii chain) and HLA-DM were previously generated. 28 HLA-DRB1*1301, 29 -DRB3*0101, 29 -DPB1*0101, 28 -DPB1*0301 7 and -DPB1*0401 30 were cloned into MP71-IRES-NGFR, whereas HLA-DQB1*0603 and HLA-DPA1*0201 28 were cloned into LZRS-IRES-GFP.…”