Expansion of T cells targeting multiple antigens of cytomegalovirus, Epstein–Barr virus and adenovirus to provide broad antiviral specificity after stem cell transplantation

• Published in: Cytotherapy (Oxford, England) England), 2011-09, Vol.13 (8), p.976-986
• Main Authors: Hanley, Patrick J, Shaffer, Donald R, Cruz, Conrad R.Y, Ku, Stephanie, Tzou, Benjamin, Liu, Hao, Demmler-Harrison, Gail, Heslop, Helen E, Rooney, Clio M, Gottschalk, Stephen, Bollard, Catherine M
• Format: Article
• Language: English
• Subjects: Adenoviridae - physiology; adenovirus; adoptive immunotherapy; Advanced Basic Science; antiviral immunity; Cell Culture Techniques; Cell Proliferation; Cells, Cultured; cytomegalovirus; Cytomegalovirus - physiology; cytotoxic T lymphocytes; Epstein–Barr virus; Hematologic Neoplasms - pathology; Hematologic Neoplasms - therapy; Hematopoietic Stem Cell Transplantation; Herpesvirus 4, Human - physiology; Humans; Immediate-Early Proteins - genetics; Immediate-Early Proteins - immunology; Immediate-Early Proteins - metabolism; Lymphocyte Activation; Other; Phosphoproteins - genetics; Phosphoproteins - immunology; Phosphoproteins - metabolism; Postoperative Complications; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - immunology; Recombinant Fusion Proteins - metabolism; stem cell transplant; T-Cell Antigen Receptor Specificity - genetics; T-Cell Antigen Receptor Specificity - immunology; T-Lymphocytes, Cytotoxic - immunology; T-Lymphocytes, Cytotoxic - metabolism; T-Lymphocytes, Cytotoxic - pathology; T-Lymphocytes, Cytotoxic - transplantation; Viral Matrix Proteins - genetics; Viral Matrix Proteins - immunology; Viral Matrix Proteins - metabolism; Virus Diseases - etiology; Virus Diseases - prevention & control
• ISSN: 1465-3249; 1477-2566; 1477-2566
• DOI: 10.3109/14653249.2011.575356

Abstract Background aims Hematopoietic stem cell transplant (HSCT) is the treatment of choice for a proportion of patients with hematologic malignancies as well as for non-malignant diseases. However, viral infections, particularly Epstein–Barr virus (EBV), cytomegalovirus (CMV) and adenovirus (Ad), remain problematic after transplant despite the use of antiviral drugs. We have shown that cytotoxic T lymphocytes (CTL) generated against CMV-pp65, EBV and Ad antigens in a single culture are capable of controlling infections with all three viruses after HSCT. Although pp65-specific CTL have proved efficacious for the control of CMV infection, several reports highlight the importance of targeting additional CMV antigens. Methods To expand multivirus-specific T cells with activity against both CMV-pp65 and CMV-IE-1, peripheral blood mononuclear cells (PBMC) were transduced with the adenoviral vector (Ad5f35-IE-1-I-pp65). After 9–12 days the CTL were restimulated with autologous EBV-transformed B cells transduced with the same Ad vector. Results After 18 days in culture nine CTL lines expanded from less than 1.5 × 107 PBMC to a mean of 6.1 × 107 T cells that recognized CMV antigens pp65 [median 273 spot-forming cells (SFC), range 47–995] and IE-1 (median 154 SFC, range 11–505), the Ad antigens hexon (median 153 SFC, range 26–465) and penton (median 37 SFC, range 1–353), as well as EBV lymphoblastoid cell lines (median 55 SFC, range 9–301). Importantly, the T cells recognized at least two antigens per virus and lysed virus peptide-pulsed targets. Conclusions CTL that target at least two antigens each of CMV, EBV and Ad should have clinical benefit with broad coverage of all three viruses and enhanced control of CMV infections compared with current protocols.