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Induction of lymphokine-activated killer activity in rat splenocyte cultures: The importance of 2-mercaptoethanol and indomethacin

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Summary

The role of 2-mercaptoethanol and indomethacin in the induction of lymphokine-activated killer (LAK) activity by interleukin-2 (IL-2) in rat splenocyte cultures was investigated. Spleens from 4-month-old male rats of five different strains were tested. Splenocytes were cultured for 3–5 days in the presence of IL-2 (1000 U/ml) and LAK activity was assessed by 4-h51Cr release assays with P815 and YAC-1 cells as targets. LAK activity could be induced by IL-2 in splenocytes from all rat strains, but only when 2-mercaptoethanol was present in the culture medium. Optimal LAK activity was induced when the 2-mercaptoethanol concentration in splenocyte cultures was at least 5 µM. Different rat strains showed differences in levels of in vitro induction of LAK activity. In the presence of 2-mercaptoethanol the level of LAK activity induced by IL-2 was high in BN and Lewis rats, intermediate in Wistar and Wag rats, and low in DZB rats. In the absence of 2-mercaptoethanol no or minimal LAK activity was induced. Furthermore we observed that addition of 50 µm indomethacin to the culture medium in the presence of 2-mercaptoethanol augmented the induction of LAK activity to some extent. In the absence of 2-mercaptoethanol, addition of indomethacin resulted only in low levels or no induction of LAK activity. We conclude that for optimal induction of LAK activity by IL-2 in rat splenocyte cultures 2-mercaptoethanol is essential, while indomethacin can only marginally further improve this induction.

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Authors and Affiliations

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

    Peter J. K. Kuppen, Emile de Heer & Gert Jan Fleuren

  2. Department of Surgery, Leiden University Hospital, 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

Authors
  1. Peter J. K. Kuppen
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  2. Alexander M. M. Eggermont
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  3. Andreas Marinelli
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  4. Emile de Heer
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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., Marinelli, A. et al. Induction of lymphokine-activated killer activity in rat splenocyte cultures: The importance of 2-mercaptoethanol and indomethacin. Cancer Immunol Immunother 33, 28–32 (1991). https://doi.org/10.1007/BF01742524

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  • DOI: https://doi.org/10.1007/BF01742524

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