Bioassaying tumor necrosis factor (TNF) relies not only on a sensitive but also on a stable cell line towards TNF action. A variant of WEHI 164 clone 13, WEHI-13VAR, was characterized by us. This variant demonstrated both stability and high sensitivity towards rTNF-alpha or rTNF-beta when assayed in the presence of actinomycin-D (AcD). Fifteen subclones were generated from WEHI-13VAR (rTNF-alpha; ED50 range: 0.005-0.065 ng/ml). No clones were found more sensitive than the AcD-treated WEHI-13VAR cell line. An important feature of this line is its stable sensitivity. The stability of the cell line sensitivity towards rTNF-beta was demonstrated in the presence of AcD over a period of 16 months with an average ED50 of 0.046 +/- 0.004 ng/ml. In conclusion, unlike other TNF-sensitive cell lines in which the sensitivity is lost or the stability is unknown, WEHI-13VAR provides a sensitive, reliable and stable bioassay system to detect cytotoxin, TNF-alpha, and TNF-beta.
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