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Adenine and hypoxanthine metabolism in phythohemagglutinin-stimulated and unstimulated human lymphocytes

Der Stoffwechsel von Adenin und Hypoxanthin in Phytohaemagglutinin-stimulierten und unstimulierten menschlichen Lymphozyten

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Zusammenfassung

Die Aufnahme und der Stoffwechsel von Adenin und Hypoxanthin in Phytohaemagglutinin-stimulierten und unstimulierten Blutlymphozyten wurden untersucht. Durch den Einbau von14C-markierten Purinen in die säurelösliche Fraktion sollte der 1. Schritt des Nukleinsäurestoffwechsels gemessen werden. Zwischen frisch präparierten und 48 h kultivierten Lymphozyten konnte keine bevorzugte Aufnahme von Adenin bzw. Hypoxanthin festgestellt werden. Die kultivierten Zellen metabolisierten jedoch im Vergleich zu den frisch gewonnenen Lymphozyten nur etwa 1/3 der Purine. Intrazellulär wurde 2/3 des Adenins und die Hälfte des Hypoxanthins hauptsächlich zu den Nukleotiden AMP und ADP metabolisiert. Eine 1stündige Inkubation mit PHA führte in den frischen Lymphozyten zu einem Anstieg der Adenin-bzw. Hypoxanthinaufnahme auf 191% bzw. 153% verglichen mit den Ausgangswerten. Hingegen stiegen die entsprechenden Werte während einer PHA-Stimulation von 48 h auf 158% bzw. 132% an. Obwohl die intrazelluläre Bildung der Nukleotide aus Adenin bzw. Hypoxanthin nach 1 h PHA-Stimulation auf 152% bzw. 161% anstieg, kam es zu keiner Änderung der relativen Durchflußraten der verschiedenen Stoffwechselwege. Im Gegensatz war nach 48 h Stimulation keine vermehrte Nukleotidbildung nachweisbar. Sowohl die gesteigerte Aufnahme als auch die erhöhte Nukleotidbildung nach 1 h PHA-Stimulation könnte auf geänderte Transporteigenschaften der Lymphozytenmembran hinweisen und die erste Antwort der Zelle auf den mitogenen Reiz darstellen. Dadurch könnte der gesteigerte Bedarf an niedermolekularen Substanzen für die nachfolgenden Biosynthesen gedeckt werden. Nach abgeschlossener Transformation würde kein vermehrter Bedarf bestehen und somit die Aufnahme und Metabolisierung gedrosselt werden.

Summary

The uptake and subsequent metabolism of adenine and hypoxanthine in phytohemagglutinin-stimulated and unstimulated peripheral human blood lymphocytes, freshly prepared or cultured, were studied. To investigate the initial step of nucleic acid metabolism the incorporation of14C-purines into the acid soluble material was examined. No preferential uptake of adenine or hypoxanthine was observed in freshly prepared and cultured lymphocytes during an incubation of 1 h. However, cultured cells utilized approximately 1/3 of the purines compared to freshly drawn cells. Within the cells 2/3 of adenine and 1/2 of hypoxanthine were metabolized to nucleotides (mainly AMP and ADP). Incubation of lymphocytes with PHA for 1 h produced in the freshly prepared cells an increase of adenine- and hypoxanthine-uptake to 191% and 153%, in 48 h stimulated cells to 158% and 132%. There was, however, no change in the relative rates of the metabolic routes though the intracellular concentrations of nucleotides formed increased with adenine as substrate to 152% and with hypoxanthine to 161% during a 1 h stimulation. In contrast no enhanced formation of acid soluble nucleotide formation could be observed with PHA stimulation during 48 h. The increased rates of purine uptake and metabolism apparent 1 h after addition of mitogen may be due to an altered transport mechanism at the beginning of the transformation as an adaptive response to the increased requirements for the synthetic processes soon to follow. Once the lymphocytes are transformed no demand of purines is necessary and the uptake and metabolism is switched off.

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Authors and Affiliations

  1. Allgemeines Krankenhaus der Stadt Wien, II. Medizinische Universitätsklinik, Garnisongasse 13, A-1090, Wien, Austria

    M. M. Müller

  2. Institut für Medizinische Chemie der Universität, A-1090, Wien, Austria

    G. Pischek

  3. Institut für allgemeine und experimentelle Pathologie der Universität, A-1090, Wien, Austria

    O. Scheiner

  4. Institut für spezifische Prophylaxe und Tropenmedizin der Universität, A-1095, Wien, Austria

    H. Stemberger & G. Wiedermann

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  1. M. M. Müller
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  2. G. Pischek
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  3. O. Scheiner
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  4. H. Stemberger
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  5. G. Wiedermann
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This work has been supported by grant No. 9796 of the “Fonds zur Förderung der wissenschaftlichen Forschung”

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Müller, M.M., Pischek, G., Scheiner, O. et al. Adenine and hypoxanthine metabolism in phythohemagglutinin-stimulated and unstimulated human lymphocytes. Blut 40, 137–145 (1980). https://doi.org/10.1007/BF01013696

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  • DOI: https://doi.org/10.1007/BF01013696

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