Ultrastructural and functional studies of the interaction between IL-12 and IL-2 for the generation of lymphokine-activated killer cells

Exp Cell Res. 1999 Dec 15;253(2):440-53. doi: 10.1006/excr.1999.4697.

Abstract

IL-12 promotes generation of LAK activity in short-term-cultured NK cells, but information on the structure and function of IL-12-induced LAK cells is not yet available. The latter issues have been here investigated with emphasis on interactions between IL-12 and IL-2. Peripheral blood mononuclear cells (MNC) exposed to IL-12 for 5-7 days displayed a decrease in the amount and density of the matrix of large granular lymphocyte (LGL)-associated granules. In cells cultured with IL-12 and IL-2 for 5-7 days, empty vacuoles were predominant and the electron-dense matrix was scanty. In MNC incubated with IL-2 for 5-7 days, most granules were loaded with electron-dense matrix. IL-12 and IL-2 displayed an additive effect on LAK cell cytotoxicity until approximately 48 h in culture which was followed by a sharp decline. Immunocytochemical and biochemical studies demonstrated that MNC cultured for 5-7 days with IL-12 and IL-2 displayed downregulated perforin expression and upregulated granzyme B expression. Fas ligand expression was virtually undetectable in MNC cultured for 5-7 days with or without cytokines. It appears that perforin downregulation plays a major role in the reduced cytotoxicity of MNC cultured with IL-12 and IL-2 for 5-7 days.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Differentiation / drug effects
  • Cell Differentiation / immunology
  • Cells, Cultured
  • Cytoplasmic Granules / immunology
  • Cytoplasmic Granules / ultrastructure
  • Fas Ligand Protein
  • Granzymes
  • Humans
  • Interleukin-12 / immunology*
  • Interleukin-12 / metabolism
  • Interleukin-12 / pharmacology
  • Interleukin-2 / immunology*
  • Interleukin-2 / metabolism
  • Interleukin-2 / pharmacology
  • Killer Cells, Natural / enzymology*
  • Killer Cells, Natural / immunology*
  • Killer Cells, Natural / ultrastructure
  • Kinetics
  • Leukocytes, Mononuclear / cytology
  • Leukocytes, Mononuclear / drug effects
  • Leukocytes, Mononuclear / immunology
  • Membrane Glycoproteins / biosynthesis
  • Membrane Glycoproteins / metabolism
  • Perforin
  • Pore Forming Cytotoxic Proteins
  • Serine Endopeptidases / biosynthesis
  • Serine Endopeptidases / metabolism
  • fas Receptor / metabolism

Substances

  • FASLG protein, human
  • Fas Ligand Protein
  • Interleukin-2
  • Membrane Glycoproteins
  • Pore Forming Cytotoxic Proteins
  • fas Receptor
  • Perforin
  • Interleukin-12
  • GZMB protein, human
  • Granzymes
  • Serine Endopeptidases