Journal ArticleDOI
Simultaneous production of interleukin 2, interleukin 4 and interferon-γ by activated human blood lymphocytes
TLDR
IL 2, IL 4 and IFN‐γ accumulated in the Golgi system, which resulted in a characteristic morphology of the staining, eliminating problems with evaluation of background signals.Abstract:
The production of interleukin 2 (IL 2), IL 4 and interferon-gamma (IFN-gamma) by in vitro activated unselected human blood mononuclear cells was studied at a single-cell level. Individual lymphokine-synthesizing cells were identified by intracellular immunofluorescent staining using cytokine-specific monoclonal or polyclonal antibodies. Cultures from adult blood donors revealed a biphasic kinetic production pattern for IL 2 and IFN-gamma with peaks occurring 4-6 and 24-30 h after initiation of the cultures. Approximately 20%-40% of the lymphocytes produced IL 2 and IFN-gamma. In contrast, only 1%-3% of the lymphocytes synthesized IL 4 with maximal frequency after 6 h of culture. CD4+ as well as CD8+ T cells contributed to the synthesis of all three lymphokines studied. CD4+CD45R- T cells were the major producers of IL 2 and IL 4, while CD8+CD45R- T cells were the most common phenotype of IFN-gamma-synthesizing cells. By performing two-color immunofluorescence studies we observed that among IL 4-producing cells every second one made simultaneously IL 2 and every fourth one made IFN-gamma. Mononuclear cells from umbilical cord blood could be stimulated to make IL 2 to the same extent as cells from adult blood donors. No IL 4 production and a strikingly reduced frequency of IFN-gamma producers were noted in cell cultures from neonates. IL 2, IL 4 and IFN-gamma accumulated in the Golgi system, which resulted in a characteristic morphology of the staining, eliminating problems with evaluation of background signals.read more
Citations
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Journal ArticleDOI
Detection of intracellular cytokines by flow cytometry
TL;DR: In this article, the carboxylic ionophore monensin was used to interrupt intracellular transport processes leading to an accumulation of the cytokine in the Golgi complex.
Journal ArticleDOI
Heterogeneity of intracellular cytokine synthesis at the single-cell level in polarized T helper 1 and T helper 2 populations.
Peter J. M. Openshaw,Erin Murphy,Nancy Ann Hosken,Vernon C. Maino,Kenneth A. Davis,Kenneth M. Murphy,Anne O'Garra +6 more
TL;DR: Results show that stimulation in the presence of polarizing stimuli results in cells producing either IFN-gamma or IL-4, but that coproduction can occur in rare cells under defined conditions.
Journal ArticleDOI
Direct demonstration of cytokine synthesis heterogeneity among human memory/effector T cells by flow cytometry.
Louis J. Picker,Manoj K. Singh,Zoran Zdraveski,John R. Treer,Sharr L. Waldrop,Paul R. Bergstresser,Paul R. Bergstresser,Vernon C. Maino +7 more
TL;DR: The data strongly support the in vivo existence of human memory/effector T-cell subsets with "preprogrammed" cytokine synthesis potential, although they suggest that these subsets may be more complex than originally proposed in the TH1/TH2 hypothesis.
Journal ArticleDOI
Interferon alpha increases the frequency of interferon gamma-producing human CD4+ T cells.
TL;DR: IFN- alpha increases the frequency of IFN-gamma-secreting CD4 Th cells and antagonizes the suppressive effect of IL-4 on IFN -gamma production, indicating that IFn-alpha may favor the induction and maintenance of Th1- like cells and thereby counteract Th2-driven allergic immune responses.
Journal ArticleDOI
Detection of intracytoplasmic cytokine using flow cytometry and directly conjugated anti-cytokine antibodies.
Calman Prussin,Dean D. Metcalfe +1 more
TL;DR: Collaborating with directly conjugated anti-cytokine antibodies becomes a specific and versatile technique for the assessment of complex cytokine production phenotypes in fresh ex vivo T cell subpopulations.
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Journal Article
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Journal Article
IL-4 is an essential factor for the IgE synthesis induced in vitro by human T cell clones and their supernatants.
G Del Prete,Enrico Maggi,Paola Parronchi,I Chretien,A Tiri,Donatella Macchia,Mario Ricci,Jacques F. Banchereau,J E de Vries,Sergio Romagnani +9 more
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