title: Expansion of T cells targeting multiple antigens of cytomegalovirus, Epstein-Barr virus and adenovirus to provide broad antiviral specificity after stem cell transplantation.
abstract: OBJECTIVE
  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 × 10(7) PBMC to a mean of 6.1 × 10(7) 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.
authors.affiliation: Texas Children's Hospital, Cell and Gene Therapy, Houston, Texas, USA.
authors.forename: Patrick J
authors.initials: PJ
authors.lastname: Hanley
authors.forename: Donald R
authors.initials: DR
authors.lastname: Shaffer
authors.forename: Conrad R Y
authors.initials: CR
authors.lastname: Cruz
authors.forename: Stephanie
authors.initials: S
authors.lastname: Ku
authors.forename: Benjamin
authors.initials: B
authors.lastname: Tzou
authors.forename: Hao
authors.initials: H
authors.lastname: Liu
authors.forename: Gail
authors.initials: G
authors.lastname: Demmler-Harrison
authors.forename: Helen E
authors.initials: HE
authors.lastname: Heslop
authors.forename: Clio M
authors.initials: CM
authors.lastname: Rooney
authors.forename: Stephen
authors.initials: S
authors.lastname: Gottschalk
authors.forename: Catherine M
authors.initials: CM
authors.lastname: Bollard
mesh_terms: D000256:Adenoviridae / Q000502:physiology*; D018929:Cell Culture Techniques; D049109:Cell Proliferation; D002478:Cells, Cultured; D003587:Cytomegalovirus / Q000502:physiology*; D019337:Hematologic Neoplasms / Q000473:pathology / Q000628:therapy*; D018380:Hematopoietic Stem Cell Transplantation; D004854:Herpesvirus 4, Human / Q000502:physiology*; D006801:Humans; D017874:Immediate-Early Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D008213:Lymphocyte Activation; D010750:Phosphoproteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D011183:Postoperative Complications; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D037182:T-Cell Antigen Receptor Specificity / Q000235:genetics / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology / Q000378:metabolism* / Q000473:pathology / Q000637:transplantation; D014763:Viral Matrix Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D014777:Virus Diseases / Q000209:etiology* / Q000517:prevention & control
chemical_list: C050678:IE1 protein, Human herpesvirus 1; D017874:Immediate-Early Proteins; D010750:Phosphoproteins; D011993:Recombinant Fusion Proteins; D014763:Viral Matrix Proteins; C065418:cytomegalovirus matrix protein 65kDa