title: Clinical-grade varicella zoster virus-specific T cells produced for adoptive immunotherapy in hemopoietic stem cell transplant recipients.
abstract: OBJECTIVE
  Varicella zoster virus (VZV) causes life-long latent infection in healthy individuals, which reactivates in 10-68% of stem cell transplant patients. Reconstituting immunity through adoptive transfer of T cells specific for VZV may aid in the prophylaxis and treatment of VZV infections. The potential for generating T cells specific for VZV using a clinically approved VZV vaccine strain was investigated.
  
  
  METHODS
  The Varivax® vaccine was used to stimulate peripheral blood mononuclear cells from healthy donors. Only reagents approved for clinical manufacture were used. Monocyte-derived dendritic cells pulsed with Varivax (R) were used to stimulate autologous mononuclear cells at a responder to stimulator ratio of 10:1. On day 7, a second stimulation was performed; 20 U/mL interleukin (IL)-2 were added from day 7 and 50 U/mL IL-2 from day 14 onwards. Cell phenotype and functionality were assessed after 21 days of culture.
  
  
  RESULTS
  A mean increase of 11-fold in cell number was observed (n= 18). Cultures were mainly T cells (mean CD3 (+) 89.7%, CD4 (+) 54.2%, CD8 (+) 28.7%) with effector and central memory phenotypes. Cells produced one or more T helper (Th)1 cytokine (interferon-γ, tumor necrosis factor-α and IL-2), and CD4 (+) (but not CD8 (+) ) cells expressed the cytoxicity marker CD107 when restimulated with VZV antigens.
  
  
  CONCLUSIONS
  We have demonstrated a clinically applicable method that yields high numbers of highly reactive T cells specific for VZV. We propose that reconstructing host immunity through adoptive transfer of VZV-specific T cells will reduce the frequency of clinical VZV infection in the period of severe immune suppression that follows allogeneic stem cell transplantation.
authors.affiliation: Westmead Millennium Institute, University of Sydney, Westmead, Australia. emily.blyth@sydney.edu.au
authors.forename: Emily
authors.initials: E
authors.lastname: Blyth
authors.forename: Shivashni S
authors.initials: SS
authors.lastname: Gaundar
authors.forename: Leighton
authors.initials: L
authors.lastname: Clancy
authors.forename: Renee M
authors.initials: RM
authors.lastname: Simms
authors.forename: Ian
authors.initials: I
authors.lastname: Bilmon
authors.forename: Kenneth P
authors.initials: KP
authors.lastname: Micklethwaite
authors.forename: David J
authors.initials: DJ
authors.lastname: Gottlieb
mesh_terms: D049109:Cell Proliferation; D003602:Cytotoxicity, Immunologic; D000939:Epitopes / Q000276:immunology; D018380:Hematopoietic Stem Cell Transplantation; D006562:Herpes Zoster / Q000276:immunology; D014645:Herpesvirus 3, Human / Q000276:immunology* / Q000472:pathogenicity; D006801:Humans; D016219:Immunotherapy, Adoptive / Q000379:methods*; D010641:Phenotype; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000276:immunology; D013045:Species Specificity; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*
chemical_list: D000939:Epitopes; D016693:Receptors, Antigen, T-Cell, alpha-beta