Abstract
Human adenovirus infection is life threatening after allogeneic haematopoietic stem cell transplantation (HSCT). Immunotherapy with donor-derived adenovirus-specific T cells is promising; however, 20% of all donors lack adenovirus-specific T cells. To overcome this, we transfected α/β T cells with mRNA encoding a T-cell receptor (TCR) specific for the HLA-A*0101-restricted peptide LTDLGQNLLY from the adenovirus hexon protein. Furthermore, since allo-reactive endogenous TCR of donor T lymphocytes would induce graft-versus-host disease (GvHD) in a mismatched patient, we transferred the TCR into γ/δ T cells, which are not allo-reactive. TCR-transfected γ/δ T cells secreted low quantities of cytokines after antigen-specific stimulation, which were increased dramatically after co-transfection of CD8α-encoding mRNA. In direct comparison with TCR-transfected α/β T cells, TCR-CD8α-co-transfected γ/δ T cells produced more tumor necrosis factor (TNF), and lysed peptide-loaded target cells as efficiently. Most importantly, TCR-transfected α/β T cells and TCR-CD8α-co-transfected γ/δ T cells efficiently lysed adenovirus-infected target cells. We show here, for the first time, that not only α/β T cells but also γ/δ T cells can be equipped with an adenovirus specificity by TCR-RNA electroporation. Thus, our strategy offers a new means for the immunotherapy of adenovirus infection after allogeneic HSCT.
Highlights
After allogeneic haematopoietic stem cell transplantation (HSCT) human adenovirus (HAdV) infection is a life threatening complication
Jurkat cells were co-electroporated with RNA encoding the T-cell receptor (TCR) and the DNA Transluc vector that encodes luciferase under the control of an NFAT-inducible promoter
HAdV/A1-TCR and CD8a co-transfected c/d T cells produce cytokines antigen- Since the endogenous TCR of the donor T lymphocytes would induce graft-versus-host disease (GvHD) in a mismatched patient, we investigated whether the HAdV/A1-specific TCR can be functionally transferred into not allo-reactive c/d T cells by mRNA electroporation. c/d T cells were isolated from expanded PBMC of healthy donors to obtain sufficient numbers for electroporation
Summary
After allogeneic haematopoietic stem cell transplantation (HSCT) human adenovirus (HAdV) infection is a life threatening complication. Immunotherapy with either magnetically separated [4] or expanded [5] HAdV-specific T cells represents a promising treatment option to overcome viral infections after allogeneic HSCT. More recent approaches are based on the short-term expansion of HAdV-specific T cells with overlapping 15-mer polypeptides from highly conserved regions of the immunodominant major capsid protein hexon [6,7], to facilitate broad recognition and protection against several HAdV species [8]. As a prerequisite for such immunotherapies, the Tcell donor has to have virus-specific T cells. Recent data from our laboratory showed that in 12 out of 50 donors, no HAdVspecific T cells were detectable via MHC class I multimers and/ or IFNc ELIspot (unpublished data). Due to the incomplete match of donor and recipient, the use of donor T cells is further restricted because they only react in the presence of matching HLA molecules
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