Tumor necrosis factor soluble receptor 75: the principal receptor form released by human alveolar macrophages and monocytes in the presence of interferon gamma

Am J Respir Cell Mol Biol. 1996 Mar;14(3):279-87. doi: 10.1165/ajrcmb.14.3.8845179.

Abstract

Tumor necrosis factor alpha(TNF alpha), a proinflammatory cytokine secreted predominantly by monocytemacrophages, interacts with two cell-surface receptors: TNF-R55 and TNF-R75. Few studies have been devoted to their modulation on human alveolar macrophages (AM). Both source and target of TNF(alpha), AM also release its inhibitors, the soluble receptors, following the cleavage of the extracellular domain of TNF-R55 and TNF-R75. Because in vivo AM are subject to activation by exogenous or endogenous stimuli, we analyzed the release of both receptors into the cell culture supernatant in response to lipopolysaccharide (LPS), phorbol myristate acetate (PMA), and cytokines such as interleukin 2(IL-2), IL-4, IL-6, granulocyte-macrophage colony-stimulating factor (GM-CSF), and interferon gamma (IFN-gamma). Results were compared with those obtained on peripheral blood monocytes (Mo), and the role of receptor recycling was investigated using inhibitors such as monensin and chloroquine. In our culture conditions, basal release by unstimulated AM amounted to 0.3 +/- 0.1 and 0.5 +/- 0.1 ng/ml for TNF-sR55 and TNF-sR75, respectively. In the same conditions, Mo released 1.2 +/- 1.2 ng/ml of TNF-sR55 and 5.1 +/- 0.1 ng/ml of TNF-sR75. PMA slightly increased mRNA expression and release of TNF-sR55, but those of TNF-sR75 were enhanced approximately 4-fold. After 24 h of culture, the release of TNF-sR75 was 2.5-fold higher on Mo than on AM. Of the cytokines tested on AM, IFN-gamma increased the release of TNF-sR75 3-fold, but that of TNF-sR55 only between 1.5- and 2-fold. GM-CSF enhanced them to a lower extent (approximately 1.5-fold). Shedding occurred despite the presence of chloroquine, monensin and colchicine, suggesting that cleavage takes place on the cell surface rather than after internalization. Addition of colchicine increased the release of TNF-sR75 induced by LPS and IFN-gamma, but not by PMA. In conclusion, Mo and AM differ in their ability to release TNF(alpha) and TNF-sR. On AM the release of each receptor appears to be regulated separately. Finally, IFN-gamma was among the most efficacious cytokines to induce the release of both receptors, with TNF-sR75 being more liable to shedding. Thus, the two TNF-R seem to be ruled by separate mechanisms and to differ in terms of release sensitivity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, CD / analysis
  • Antigens, CD / metabolism*
  • Cells, Cultured
  • Chloroquine / pharmacology
  • Colchicine / pharmacology
  • Culture Media
  • Growth Substances / pharmacology
  • Humans
  • Interferon-gamma / pharmacology*
  • L Cells
  • Lipopolysaccharides / pharmacology
  • Macrophage Activation
  • Macrophages, Alveolar / metabolism*
  • Mice
  • Microtubules / drug effects
  • Monensin / pharmacology
  • Monocytes / metabolism*
  • Protein Synthesis Inhibitors / pharmacology
  • RNA, Messenger / biosynthesis
  • Receptors, Tumor Necrosis Factor / analysis
  • Receptors, Tumor Necrosis Factor / metabolism*
  • Receptors, Tumor Necrosis Factor, Type I
  • Receptors, Tumor Necrosis Factor, Type II
  • Tetradecanoylphorbol Acetate / pharmacology
  • Tumor Necrosis Factor-alpha / analysis
  • Tumor Necrosis Factor-alpha / biosynthesis
  • Tumor Necrosis Factor-alpha / metabolism

Substances

  • Antigens, CD
  • Culture Media
  • Growth Substances
  • Lipopolysaccharides
  • Protein Synthesis Inhibitors
  • RNA, Messenger
  • Receptors, Tumor Necrosis Factor
  • Receptors, Tumor Necrosis Factor, Type I
  • Receptors, Tumor Necrosis Factor, Type II
  • Tumor Necrosis Factor-alpha
  • Interferon-gamma
  • Chloroquine
  • Monensin
  • Tetradecanoylphorbol Acetate
  • Colchicine