Development in vitro of human CD4+ thymocytes into functionally mature Th2 cells. Exogenous interleukin-12 is required for priming thymocytes to produce both Th1 cytokines and interleukin-10

Maria Cristina Mingari, Enrico Maggi, Anna Cambiaggi, Francesco Annunziato, Francesca Schiavetti, Roberto Manetti, Lorenzo Moretta, Sergio Romagnani

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

Fresh postnatal thymocyte cell suspensions were directly cloned under limiting dilution conditions with either phytohemagglutinin or toxic shock syndrome toxin-1 (TSST-1), a bacterial superantigen. Cultures contained allogenic irradiated feeder cells and interleukin (IL)-2, in the absence or presence of exogenous IL-4, interferon (IFN)-γ or IL-12. The resulting CD4+ T cell clones generated under these different experimental conditions were then analyzed for their ability to produce IL-2, IL-4, IL-5, IL-10, IFN-γ and tumor necrosis factor (TNF)-β in response to stimulation with phorbol 12-myristate 13-acetate (PMA)+anti-CD3 monoclonal antibody or PMA + ionomycin. Different from T cell clones generated from peripheral blood, virtually all CD4+ T cell clones generated from human thymocytes produced high concentrations of IL-2, IL-4 and IL-5, but no IFN-γ, TNF-β or IL-10. Moreover, after activation, these clones expressed on their surface membrane both CD30 and CD40 ligand, but not the product of lymphocyte activation gene (LAG)-3, and provided strong helper activity for IgE synthesis by allogeneic B cells. The Th2 cytokine pattern could not be modified by the addition of IFN-γ. However, upon addition of exogenous IL-12, the resulting CD4+ thymocyte clones produced TNF-β, IFN-γ, and IL-10 in addition to IL-4 and IL-5. These results suggest that CD4 human thymocytes have the potential to develop into cells producing the Th2 cytokines IL-4 and IL-5, whereas the ability to produce both Th1 cytokines and IL-10 is acquired only after priming with IL-12.

Original languageEnglish
Pages (from-to)1083-1087
Number of pages5
JournalEuropean Journal of Immunology
Volume26
Issue number5
Publication statusPublished - May 1996

Fingerprint

Th2 Cells
Thymocytes
Interleukin-12
Interleukin-4
Interleukin-10
Interferons
Interleukin-5
Clone Cells
Cytokines
Interleukin-2
Tumor Necrosis Factor-alpha
T-Lymphocytes
CD30 Ligand
Acetates
Feeder Cells
Superantigens
CD40 Ligand
Ionomycin
Phytohemagglutinins
Lymphocyte Activation

Keywords

  • Interleukin-12 priming
  • T cell clone
  • Th2 cell
  • Thymus

ASJC Scopus subject areas

  • Immunology

Cite this

Development in vitro of human CD4+ thymocytes into functionally mature Th2 cells. Exogenous interleukin-12 is required for priming thymocytes to produce both Th1 cytokines and interleukin-10. / Mingari, Maria Cristina; Maggi, Enrico; Cambiaggi, Anna; Annunziato, Francesco; Schiavetti, Francesca; Manetti, Roberto; Moretta, Lorenzo; Romagnani, Sergio.

In: European Journal of Immunology, Vol. 26, No. 5, 05.1996, p. 1083-1087.

Research output: Contribution to journalArticle

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abstract = "Fresh postnatal thymocyte cell suspensions were directly cloned under limiting dilution conditions with either phytohemagglutinin or toxic shock syndrome toxin-1 (TSST-1), a bacterial superantigen. Cultures contained allogenic irradiated feeder cells and interleukin (IL)-2, in the absence or presence of exogenous IL-4, interferon (IFN)-γ or IL-12. The resulting CD4+ T cell clones generated under these different experimental conditions were then analyzed for their ability to produce IL-2, IL-4, IL-5, IL-10, IFN-γ and tumor necrosis factor (TNF)-β in response to stimulation with phorbol 12-myristate 13-acetate (PMA)+anti-CD3 monoclonal antibody or PMA + ionomycin. Different from T cell clones generated from peripheral blood, virtually all CD4+ T cell clones generated from human thymocytes produced high concentrations of IL-2, IL-4 and IL-5, but no IFN-γ, TNF-β or IL-10. Moreover, after activation, these clones expressed on their surface membrane both CD30 and CD40 ligand, but not the product of lymphocyte activation gene (LAG)-3, and provided strong helper activity for IgE synthesis by allogeneic B cells. The Th2 cytokine pattern could not be modified by the addition of IFN-γ. However, upon addition of exogenous IL-12, the resulting CD4+ thymocyte clones produced TNF-β, IFN-γ, and IL-10 in addition to IL-4 and IL-5. These results suggest that CD4 human thymocytes have the potential to develop into cells producing the Th2 cytokines IL-4 and IL-5, whereas the ability to produce both Th1 cytokines and IL-10 is acquired only after priming with IL-12.",
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AU - Cambiaggi, Anna

AU - Annunziato, Francesco

AU - Schiavetti, Francesca

AU - Manetti, Roberto

AU - Moretta, Lorenzo

AU - Romagnani, Sergio

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