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Soluble factors produced by macrophages/monocytes inhibit lymphokine-activated killer activity in rat splenocyte cultures

  • Original Articles
  • LAK, Interleukin-2, 2-Mercaptoenthanol, Macrophages/Monocytes, Rat
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Abstract

In the present study we investigated the inhibition of interleukin-2(IL-2)-induced lymphokine-activated killer (LAK) activity in rat splenocyte cultures in relation to the presence of 2-mercaptoethanol and macrophages/monocytes. The presence of 2-mercaptoethanol is necessary for induction of LAK activity in rat splenocyte cultures. Removal of macrophages/monocytes from rat splenocytes by plastic or nylon-wool adherence, or iron ingestion resulted in LAK induction by IL-2 in the absence of 2-mercaptoethanol. The effect of macrophages/monocytes on LAK activity was also studied in transwell co-cultures. In the absence of 2-mercaptoethanol, the induction of LAK activity was very low in macrophage/monocyte-depleted splenocytes with macrophages/monocytes in the upper compartment of a transwell culture. In contrast, in the presence of 2-mercaptoethanol a high level of LAK activity was induced in these transwell cultures, showing that 2-mercaptoethanol abolished the LAK-inhibiting capacity of macrophages/monocytes. In addition, established LAK activity was strongly inhibited when, after LAK induction, splenocytes were cultured with supernatant of unfractionated splenocytes, which were cultured with IL-2 but in the absence of 2-mercaptoethanol. Addition of 2-mercaptoethanol abrogated the inhibiting effect of the supernatant completely. These experiments demonstrate that rat macrophages/monocytes produce 2-mercaptoethanolsensitive soluble LAK-inhibiting factors. Ultrafiltration of conditioned culture medium of macrophages/monocytes revealed the presence of LAK-inhibiting factors larger than 10 kDa. We concluded that 2-mercaptoethanol-sensitive soluble factors produced by macrophages/monocytes determine the level of LAK induction in rat splenocyte cultures.

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Authors and Affiliations

  1. Department of Pathology, University of Leiden, P.O. Box 9603, 2300RC, Leiden, The Netherlands

    Peter J. K. Kuppen, Rianne B. W. M. Quak & Gert Jan Fleuren

  2. Department of Surgery, University of Leiden, Leiden, The Netherlands

    Peter J. K. Kuppen, Andreas Marinelli & Cornelis J. H. van de Velde

  3. Department of Surgery, Rotterdam Cancer Institute, Rotterdam, The Netherlands

    Alexander M. M. Eggermont

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  1. Peter J. K. Kuppen
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  2. Alexander M. M. Eggermont
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  3. Rianne B. W. M. Quak
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  4. Andreas Marinelli
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  5. Cornelis J. H. van de Velde
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  6. Gert Jan Fleuren
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Kuppen, P.J.K., Eggermont, A.M.M., Quak, R.B.W.M. et al. Soluble factors produced by macrophages/monocytes inhibit lymphokine-activated killer activity in rat splenocyte cultures. Cancer Immunol Immunother 38, 61–67 (1994). https://doi.org/10.1007/BF01517171

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  • DOI: https://doi.org/10.1007/BF01517171

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