Comparison of the responses of freshly isolated and cultured human monocytes and P388D1 cells to agents affecting cyclic AMP metabolism

V. R. Lavis, S. J. Strada, C. P. Ross, E. M. Hersh, W. J. Thompson

Research output: Contribution to journalArticle

11 Scopus citations

Abstract

We have studied the effects of beta-adrenergic agonists, PGE1, MIX, and insulin on the metabolism and function of freshly isolated and cultured human monocytes and P388D1 cells. Human monocytes were isolated from fresh defibrinated blood by centrifugation through Ficoll/Hypaque, followed by adherence to plastic tissue culture dishes. Immediately after the adherence step, basal monocytic cyclic AMP was 0.61 ± 0.06 pmol/106 cells (mean ± S.E. for eight different preparations). The cyclic AMP content of fresh monocytes was transiently increased no more than fivefold by incubation with 0.1 mM ISO together with 0.1 mM MIX. Cellular cyclic AMP was elevated 70-fold by 10 μM PGE1 with 0.1 mM MIX. The basal cyclic AMP of monocytes cultured for 7 days was 3.77 ± 0.86 pmol/106 cells (seven preparations). Their responses to ISO, MIX, and PGE1 were similar to those of fresh monocytes. Insulin (0.2 μM) had no effect on cyclic AMP of either fresh or cultured monocytes. The cyclic AMP content of “macrophage-like” P388D1 cells grown in monolayer culture was elevated ninefold by 10 μM PGE1 and 50-fold by 10 μM PGE1 plus 0.1 mM MIX. However, 0.1 mM ISO did not increase cyclic AMP, even in the presence of MIX. Insulin (0.2 μM) did not change cyclic AMP in the presence or absence of PGE1. In sonicated P388D1 cells, adenylyl cyclase activity was increased fivefold by 10 μM PGE1 but was insensitive to 0.5 mM EPI, 0.5 mM ISO, or 0.2 μM insulin. Insulin (0.1 μM) had no effect on incorporation of [14C]glucose into glycogen by any of the cell types studied. Also, insulin (0.25 nM to 1.25 μM) did not influence performance of ADCC by human monocytes. We conclude that the adenylyl cyclases of human monocytes and P388D1 cells respond strongly to PGE1 but weakly to beta-adrenergic agonists. Since there was no effect of insulin on monocytic cyclic AMP, glycogen metabolism, or antibody-dependent effector function, we suggest that human monocytic insulin-binding sites are not coupled to physiologically important effector systems.

Original languageEnglish (US)
Pages (from-to)551-561
Number of pages11
JournalJournal of Laboratory and Clinical Medicine
Volume96
Issue number3
StatePublished - Jan 1 1980

Keywords

  • 1-methyl-3-isobutylxanthine
  • 5′-cyclic monophosphate [H]cAMP
  • 5′-guanylylimidodiphosphate
  • ADCC
  • BCG
  • EPI
  • Eagle's minimum essential medium
  • Gpp(NH)p
  • ISO
  • MEM
  • MIX
  • PGE
  • [G-H]adenosine-3′
  • antibody-dependent cell-mediated cytotoxicity
  • bacille Calmette-Guérin
  • l-epinephrine
  • l-isoproterenol
  • prostaglandin E

ASJC Scopus subject areas

  • Pathology and Forensic Medicine

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