Functional CD4(+) and CD8(+) T-cell responses induced by autologous mitomycin C treated Epstein-Barr virus transformed lymphoblastoid cell lines

P Meij, E Bloemena, N Palmen, A Brink, M B Vervoort, C J Meijer, J M Middeldorp

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6 Citations (Scopus)


Epstein-Barr virus (EBV) gene expression in tumor cells of posttransplant lymphoproliferative disorder (PTLD) patients resembles that of EBV transformed B-cell lines (LCL). EBV-specific cytotoxic T-lymphocytes can be generated by stimulating peripheral blood lymphocytes with autologous LCL. We describe a standardized method for the growth inactivation and cryopreservation of LCL for optimal T-cell stimulation and analyzed the function and phenotype of responding T-cells. LCL growth was completely blocked by mitomycin C treatment (McLCL) and McLCL could be cryopreserved while retaining excellent APC function. McLCL stimulated both CD4(+) and CD8(+) T-cells as measured by HLA-DR and CD25 expression using FACS analysis. EBV-specific CTL activity and T-cell proliferation were induced and immunocytochemical staining showed CD4(+) and (granzyme B positive) CD8(+) T-cells rosetting with McLCL. Granzymes A and B, IFN-gamma, and IL-6 were detected at significant levels in the supernatant. Thus, ex vivo T-cell activation with cryopreserved McLCL results in activation of both CD4(+) and CD8(+) T-cells producing a Th1-like cytokine profile, making this a suitable protocol for adoptive therapy of PTLD.

Original languageEnglish
Pages (from-to)25-33
Number of pages9
JournalCellular Immunology
Issue number1
Publication statusPublished - 25 Feb 2001


  • Adoptive Transfer
  • Antigen Presentation
  • CD4-Positive T-Lymphocytes
  • CD8-Positive T-Lymphocytes
  • Cell Division
  • Cell Line, Transformed
  • Cryopreservation
  • Cytotoxicity, Immunologic
  • Enzyme-Linked Immunosorbent Assay
  • Flow Cytometry
  • Granzymes
  • Herpesvirus 4, Human
  • Humans
  • Interferon-gamma
  • Interleukin-6
  • Journal Article
  • Lymphocyte Activation
  • Mitomycin
  • Phenotype
  • Rosette Formation
  • Serine Endopeptidases
  • T-Lymphocytes, Cytotoxic
  • Tumor Cells, Cultured

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