title: Evaluation of human cytomegalovirus-specific CD8+ T-cells in allogeneic haematopoietic stem cell transplant recipients using pentamer and interferon-γ-enzyme-linked immunospot assays.
abstract: BACKGROUND
  Reactivation of latent human cytomegalovirus (HCMV) is a frequent complication following allogeneic haematopoietic stem cell transplantation (HSCT). Evaluation of the quantity and function of HCMV-specific CD8+ T-cell responses after HSCT may play a crucial role in the prevention of HCMV reactivation.
  
  
  OBJECTIVE
  To investigate the mechanism of HCMV-specific T-cell immune responses after HSCT in HCMV-specific CD8+ T cells.
  
  
  METHODS
  HCMV-specific CD8+ T cells were quantified using human leucocyte antigen (HLA) pentamer staining and functionally analysed by interferon-γ-enzyme-linked immunospot (IFN-γ-ELISPOT) assay with a pp65495-503 peptide in recipients four years after HSCT.
  
  
  RESULTS
  The absolute number of pp65495-503-specific CD8+ T cells did not differ significantly (p>0.05) between samples with antigenaemia and those without antigenaemia given a mean of 54.5/μl and 40.5/μl, respectively, in 21 HLA-A* 0201 patients after HSCT. The level of pp65495-503-specific CD8+ T cells>20/μl of peripheral blood was maintained 90 days after transplantation. There was a significant difference in the spot count of IFN-γ-secreting T cells between samples with antigenaemia (mean, 507/2.5×10(5)PBMCs) and those without antigenaemia (mean, 216/2.5×10(5)PBMCs; p<0.05).
  
  
  CONCLUSIONS
  pp65495-503-specific CD8+ T cells may not be sufficient to control HCMV reactivation in recipients after HSCT. However, the combination of pentamer and IFN-γ-ELISPOT assays may be valuable for evaluating HCMV-specific CD8+ T cells. Further studies on HCMV-specific T-cell immune responses continue to be performed for the prevention of persistent HCMV reactivation.
authors.affiliation: State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, School of Medicine, Zhejiang University, 79 Qingchun Road, Hangzhou 310003, China.
authors.forename: Anbing
authors.initials: A
authors.lastname: Liu
authors.forename: Yadan
authors.initials: Y
authors.lastname: Ma
authors.forename: Wei
authors.initials: W
authors.lastname: Wu
authors.forename: Xiaoming
authors.initials: X
authors.lastname: Chen
authors.forename: Yaping
authors.initials: Y
authors.lastname: Huang
authors.forename: Jianhua
authors.initials: J
authors.lastname: Hu
authors.forename: Hanying
authors.initials: H
authors.lastname: Liang
authors.forename: Huiqi
authors.initials: H
authors.lastname: Wang
authors.forename: Rong
authors.initials: R
authors.lastname: Yang
authors.forename: Jun
authors.initials: J
authors.lastname: Fan
mesh_terms: D000293:Adolescent; D000328:Adult; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D003587:Cytomegalovirus / Q000276:immunology*; D058501:Enzyme-Linked Immunospot Assay; D005260:Female; D006680:HLA Antigens / Q000032:analysis*; D018380:Hematopoietic Stem Cell Transplantation / Q000009:adverse effects*; D006801:Humans; D007371:Interferon-gamma / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D014180:Transplantation; D055815:Young Adult
chemical_list: D006680:HLA Antigens; D007371:Interferon-gamma
keywords: AA; ALL; AML; ARA-C; ATG; Allogeneic haematopoietic stem cell transplantation; Antigenaemia; BU; CD8+ T cells; CML; CSA; CTLs; CY; ELISPOT; FLU; GVHD; HCMV; HLA; HSCT; Human cytomegalovirus; IFN-γ; MDS; MMF; MP; MTX; MeCCNU; PBMCs; PRED; Pentamer; acute lymphoblastic leukaemia; acute myeloid leukaemia; antithymocyte globulin; aplastic anaemia; busulfan; chronic myeloid leukaemia; cyclophosphamide; cyclosporin A; cytosine arabinoside; cytotoxic T lymphocytes; enzyme-linked immunospot; fludarabine; graft-versus-host disease; haematopoietic stem cell transplantation; human cytomegalovirus; human leucocyte antigen; methotrexate; methylcyclohexylnitrosamine; methylprednisone; mycophenolate mofetil; myelodysplastic syndrome; peripheral blood mononuclear cells; prednisone