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A comparison between two methods for measuring tumor necrosis factor in biological fluids

I M Garrelds1, F J Zijlstra, C J Tak

  • 1Dept. of Pharmacology, Erasmus University, Rotterdam, The Netherlands.

Agents and Actions
|January 1, 1993
PubMed
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The MTT assay is a more sensitive and simpler method for measuring tumor necrosis factor-alpha (TNF-alpha) compared to the 3H-thymidine assay. This finding aids in the efficient detection of anti-tumor activity in biological fluids.

Area of Science:

  • Immunology
  • Cell Biology
  • Biochemistry

Background:

  • Tumor necrosis factor-alpha (TNF-alpha) is a crucial cytokine involved in immune responses and inflammation.
  • Accurate measurement of TNF-alpha activity in biological fluids is essential for research and diagnostics.
  • Existing methods for TNF-alpha measurement vary in sensitivity, simplicity, and cost.

Purpose of the Study:

  • To compare the efficiency of the MTT tetrazolium cytotoxic assay and the 3H-thymidine incorporation cytostatic assay for measuring TNF-alpha.
  • To evaluate the sensitivity and simplicity of these assays for detecting anti-tumor activity.

Main Methods:

  • Comparison of MTT assay and 3H-thymidine assay using human recombinant TNF-alpha, human colonic tissue, and stimulated macrophages.
  • Utilized murine myelomonocytic leukaemia WEHI-164 and L-929-transformed murine fibroblast cell-lines for the MTT assay.

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  • Employed the L-929 cell-line for the 3H-thymidine assay.

    • Main Results:

    • The MTT assay demonstrated higher sensitivity to TNF-alpha compared to the 3H-thymidine assay.
    • The WEHI-164 cell-line showed greater sensitivity than the L-929 cell-line in the MTT assay.
    • Both assays can detect anti-tumor activity, but the MTT assay proved more sensitive and simpler.

    Conclusions:

    • The MTT assay is a preferred method for measuring TNF-alpha due to its superior sensitivity and simplicity.
    • This assay facilitates reliable and cost-effective detection of anti-tumor activity in biological samples.
    • The MTT assay offers an advantageous approach for studying TNF-alpha's role in various biological contexts.