Comparison of the responses of freshly isolated and cultured human monocytes and P388D 1 cells to agents affecting cyclic AMP metabolism

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

Research output: Contribution to journalArticle

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Abstract

The authors have studied the effects of beta-adrenergic agonists, PGE 1, MIX, and insulin on the metabolism and function of freshly isolated and cultured human monocytes and P388D 1 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/10 6 cells (mean ± S.E. for 8 different preparations). The cyclic AMP content of fresh monocytes was transiently increased no more than 5-fold by incubation with 0.1 mM ISO together with 0.1 mM MIX. Cellular cyclic AMP was elevated 70-fold by 10 μM PGE 1 with 0.1 mM MIX. The basal cyclic AMP of monocytes cultured for 7 days was 3.77 ± 0.86 pmol/10 6 cells (7 preparations). Their responses to ISO, MIX, and PGE 1 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' P388D 1 cells grown in monolayer culture was elevated 9-fold by 10 μM PGE 1 and 50-fold by 10 μM PGE 1 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 PGE 1. In sonicated P388D 1 cells, adenylyl cyclase activity was increased 5-fold by 10 μM PGE 1 but was insensitive to 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. The authors conclude that the adenylyl cyclases of human monocytes and P388D 1 cells respond strongly to PGE 1 but weakly to beta-adrenergic agonists. Since there was no effect of insulin on monocytic cyclic AMP, glycogen metabolism, or antibody-dependent effector function, the authors 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
JournalThe Journal of Laboratory and Clinical Medicine
Volume96
Issue number3
StatePublished - 1980
Externally publishedYes

Fingerprint

Metabolism
Cyclic AMP
Monocytes
Insulin
Prostaglandins E
Adrenergic beta-Agonists
Glycogen
Adenylyl Cyclases
Adrenergic beta-1 Receptor Agonists
Diatrizoate
Antibody-Dependent Cell Cytotoxicity
Ficoll
Tissue culture
Centrifugation
Macrophages
Cell culture
Monolayers
Blood
Binding Sites
Cells

ASJC Scopus subject areas

  • Medicine(all)
  • Pathology and Forensic Medicine

Cite this

Comparison of the responses of freshly isolated and cultured human monocytes and P388D 1 cells to agents affecting cyclic AMP metabolism. / Lavis, V. R.; Strada, S. J.; Ross, C. P.; Hersh, Evan M; Thompson, W. J.

In: The Journal of Laboratory and Clinical Medicine, Vol. 96, No. 3, 1980, p. 551-561.

Research output: Contribution to journalArticle

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AU - Hersh, Evan M

AU - Thompson, W. J.

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N2 - The authors have studied the effects of beta-adrenergic agonists, PGE 1, MIX, and insulin on the metabolism and function of freshly isolated and cultured human monocytes and P388D 1 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/10 6 cells (mean ± S.E. for 8 different preparations). The cyclic AMP content of fresh monocytes was transiently increased no more than 5-fold by incubation with 0.1 mM ISO together with 0.1 mM MIX. Cellular cyclic AMP was elevated 70-fold by 10 μM PGE 1 with 0.1 mM MIX. The basal cyclic AMP of monocytes cultured for 7 days was 3.77 ± 0.86 pmol/10 6 cells (7 preparations). Their responses to ISO, MIX, and PGE 1 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' P388D 1 cells grown in monolayer culture was elevated 9-fold by 10 μM PGE 1 and 50-fold by 10 μM PGE 1 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 PGE 1. In sonicated P388D 1 cells, adenylyl cyclase activity was increased 5-fold by 10 μM PGE 1 but was insensitive to 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. The authors conclude that the adenylyl cyclases of human monocytes and P388D 1 cells respond strongly to PGE 1 but weakly to beta-adrenergic agonists. Since there was no effect of insulin on monocytic cyclic AMP, glycogen metabolism, or antibody-dependent effector function, the authors suggest that human monocytic insulin-binding sites are not coupled to physiologically important effector systems.

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