Generation of high quantities of viral and tumor-specific human CD4 + and CD8 + T-cell clones using peptide pulsed mature dendritic cells

Jean François Fonteneau, Marie Larsson, Selin Somersan, Catherine Sanders, Christian Münz, William W. Kwok, Nina Bhardwaj, Francine Jotereau

Research output: Contribution to journalArticlepeer-review

82 Scopus citations

Abstract

CD4 + and CD8 + T cells are key components of immune response against tumors and viruses. Many techniques have been used to clone and expand these cells in vitro for purposes of immunotherapy. Here, we describe an improved method to obtain large quantities of tumor and virus-specific human CD4 + and CD8 + T-cell clones. T cells derived from peripheral blood mononuclear cells (PBMCs) of healthy donors were stimulated several times by peptide pulsed monocyte-derived mature dendritic cells (DCs) in the presence of exogenous cytokines. T cells specific for influenza or melanoma antigens were detected by IFN-γ intracellular staining and were cloned by limiting dilution. Specific polyclonal T-cell populations were derived for all epitopes presented by mature DCs. Nine different populations were cloned and clones were raised from eight of them. Clonality was verified by HLA/peptide tetramer staining. With additional rounds of stimulation after the cloning procedure, it was possible to obtain from 109 to 1012 of each clone. Furthermore, clones could be maintained in culture in the presence of IL-2 for at least 1 month without losing their antigen-specific reactivity (e.g. cytokine secretion, cytolytic activity and proliferation). Importantly, a majority of the CD8 + T-cell clones recognized endogenously processed antigens. This method is of value for the purposes of adoptive anti-virus or anti-tumor immunotherapy.

Original languageEnglish
Pages (from-to)111-126
Number of pages16
JournalJournal of Immunological Methods
Volume258
Issue number1-2
DOIs
StatePublished - 1 Dec 2001
Externally publishedYes

Keywords

  • Adoptive immunotherapy
  • Dendritic cells
  • Human T-cell clones
  • Viral and tumor immunity

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