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The immunosuppressive substance 2-chloro-2-deoxyadenosine modulates lipoprotein metabolism in a murine macrophage cell line (P388 cells)

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Lipids

Abstract

A recently developed immunosuppressive substance, 2-chloro-2-deoxyadenosine (2-CdA), was reported to inhibit monocyte functions at low concentration. Because macrophages play a key role in the formation of atherosclerotic plaques, it was of interest to study the effect of 2-CdA on cellular lipid metabolism. For this purpose we have used a macrophage cell line (P388) to perform incubation studies in the presence of acetylated low density lipoprotein (Ac-LDL) and 2-CdA. The addition of 2-CdA, in concentrations ranging from 5–20 nM, induced a dose-dependent decrease in cellular cholesterol content and in the amount of extracellular [14C]oleic acid (OA) incorporated into the cholesteryl ester (CE) fraction. The effect was maximized at 20 nM 2-CdA with an 86% reduction in cholesterol esterification compared to controls (P<0.008). To evaluate the mechanism of interaction of 2-CdA with cellular lipid metabolism, deoxycytidine (dCyt) and 3-methoxybenzamide (3-MOB), substances known to antagonize the effect of 2-CdA in different ways, were co-administered with 2-CdA. dCyt, a competitive inhibitor of dCyt kinase, which catalyzes phosphorylation to the active metabolite, antagonized the effects of 20 nM 2-CdA, producing significantly greater incorporation of extracellular [14C]OA into the CE fraction than in the presence of 2-CdA alone (P<0.0086). Co-incubation with 2-CdA and the poly-ADP-ribose synthetase inhibitor 3-MOB, which is known to render cells resistant to 2-CdA toxicity by preventing cellular nicotinamide adenine dinucleotide (NAD)- and adenosine triphosphase-depletion, also reversed the effect of 2-CdA on lipid accumulation. However, incubation of P388 cells with 20 nM 2-CdA did not result in a decrease in cellular NAD content. As 20 nM 2-CdA showed no effect on intracellular cholesterol synthesis based on measurement by 3-hydroxy-3-methylglutaryl coenzyme A reductase activity, the decrease in cellular cholesterol content and in [14C]OA incorporation seems to be primarily due to an interference with Ac-LDL metabolism.

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Abbreviations

ACAT:

acyl coenzyme A:cholesterol acyltransferase

Ac-LDL:

acetylated low density lipoprotein

ADP:

adenosine diphosphate

AMP:

adenosine-monophosphate

2-CdA:

2-chloro-2-deoxyadenosine

cdATP:

chlorodeoxy adenosine triphosphate

CE:

cholesteryl ester

dCyt:

deoxycytidine

EDTA:

ethylenediaminetetraacetic acid

HMG-CoA:

3-hydroxy-3-methylglutaryl coenzyme A

LDL:

low density lipoprotein

3-MOB:

3-methoxybenzamide

NAD:

nicotinamide adenine dinucleotide

OA:

oleic acid

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Authors and Affiliations

  1. Department of Internal Medicine, University of Innsbruck, Anichstrasse 35, A-6020, Innsbruck, Austria

    Monika Lechleitner, Udo Zilian, Fritz Hoppichler, Michael Schirmer, Bernhard Föger, Francoise Geisen, Josef R. Patsch & Günther Konwalinka

  2. Department of Biochemistry, University of Innsbruck, A-6020, Innsbruck, Austria

    Bernhard Auer

Authors
  1. Monika Lechleitner
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  2. Bernhard Auer
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  8. Josef R. Patsch
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  9. Günther Konwalinka
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Lechleitner, M., Auer, B., Zilian, U. et al. The immunosuppressive substance 2-chloro-2-deoxyadenosine modulates lipoprotein metabolism in a murine macrophage cell line (P388 cells). Lipids 29, 627–633 (1994). https://doi.org/10.1007/BF02536097

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  • DOI: https://doi.org/10.1007/BF02536097

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