Simultaneous production of interleukin 2, interleukin 4 and interferon-gamma by activated human blood lymphocytes

Eur J Immunol. 1990 Jul;20(7):1591-6. doi: 10.1002/eji.1830200727.


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Antibodies, Monoclonal
  • Cytoplasm / analysis
  • Fluorescent Antibody Technique
  • Humans
  • In Vitro Techniques
  • Infant, Newborn
  • Interferon-gamma / biosynthesis*
  • Interleukin-2 / biosynthesis*
  • Interleukin-4 / biosynthesis*
  • Ionomycin / pharmacology
  • Kinetics
  • Leukocytes, Mononuclear / metabolism*
  • Lymphocyte Activation / drug effects
  • Lymphocyte Activation / physiology
  • Phenotype
  • Tetradecanoylphorbol Acetate / pharmacology


  • Antibodies, Monoclonal
  • Interleukin-2
  • Interleukin-4
  • Ionomycin
  • Interferon-gamma
  • Tetradecanoylphorbol Acetate