Ovalbumin sensitization of guinea-pigs reduces fMLP-induced calcium signal in alveolar macrophages

Life Sci. 2001 Aug 24;69(14):1597-607. doi: 10.1016/s0024-3205(01)01239-5.

Abstract

In this study we analyzed the N-formyl-Met-Leu-Phe (fMLP)-induced calcium signal in alveolar macrophages (AM) isolated from ovalbumin-sensitized (OA-sensitized AM) and naive (naive AM) guinea-pigs. Guinea-pigs were sensitized by subcutaneous injection of OA and AM were isolated by bronchoalveolar lavage 6 weeks thereafter. On the following day, we measured in resting and fMLP-stimulated cells: intracellular calcium concentration by fura-2 imaging analysis, forskolin-induced cyclic AMP production and superoxide dismutase inhibitable superoxide anion release of adherent AM. Resting calcium was 82+/-5.0 nM (n=217) and 144+/-9.3 nM (n=213, P<0.001) in naive and OA-sensitized AM respectively. fMLP (10(-11)-10(-7)M) induced a dose-dependent calcium increase, 10(-8)M being the maximal effective dose in both naive and OA-sensitized AM. However, at all doses tested, this fMLP effect was lower in OA-sensitized than in naive AM. While in resting condition 10(-5)M forskolin increased cyclic AMP both in naive and OA-sensitized AM, in fMPL-stimulated AM forskolin was effective only in OA-sensitized AM. Superoxide anion release measured 10 min after fMLP stimulus was higher in naive than in sensitized AM. These data suggest that the fMLP-induced intracellular signal is different in OA-sensitized AM compared to naive cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenylyl Cyclases / metabolism
  • Animals
  • Calcium / metabolism
  • Calcium Signaling / drug effects*
  • Colforsin / pharmacology
  • Cyclic AMP / biosynthesis
  • Enzyme Activation
  • Fluorescent Dyes
  • Fura-2
  • Guinea Pigs
  • Macrophages, Alveolar / drug effects*
  • Macrophages, Alveolar / enzymology
  • Macrophages, Alveolar / metabolism
  • Male
  • N-Formylmethionine Leucyl-Phenylalanine / pharmacology*
  • Ovalbumin / administration & dosage*
  • Superoxide Dismutase / metabolism
  • Superoxides / metabolism
  • Virulence Factors, Bordetella / pharmacology

Substances

  • Fluorescent Dyes
  • Virulence Factors, Bordetella
  • Superoxides
  • Colforsin
  • N-Formylmethionine Leucyl-Phenylalanine
  • Ovalbumin
  • Cyclic AMP
  • Superoxide Dismutase
  • Adenylyl Cyclases
  • Calcium
  • Fura-2