Khabar K S, Siddiqui S, Armstrong J A
Department of Biological and Medical Research, King Faisal Specialist Hospital, Riyadh, Saudi Arabia.
Immunol Lett. 1995 May;46(1-2):107-10. doi: 10.1016/0165-2478(95)00026-2.
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.
肿瘤坏死因子(TNF)的生物测定不仅依赖于对TNF作用敏感的细胞系,还依赖于稳定的细胞系。我们对WEHI 164克隆13的一个变体WEHI-13VAR进行了表征。当在放线菌素-D(AcD)存在的情况下进行检测时,该变体对重组TNF-α(rTNF-α)或重组TNF-β(rTNF-β)表现出稳定性和高敏感性。从WEHI-13VAR(rTNF-α;半数有效剂量范围:0.005 - 0.065 ng/ml)产生了15个亚克隆。未发现比经AcD处理的WEHI-13VAR细胞系更敏感的克隆。该细胞系的一个重要特征是其稳定的敏感性。在AcD存在的情况下,该细胞系对rTNF-β的敏感性稳定性在16个月的时间内得到了证明,平均半数有效剂量为0.046±0.004 ng/ml。总之,与其他敏感性丧失或稳定性未知的TNF敏感细胞系不同,WEHI-13VAR提供了一个灵敏、可靠且稳定的生物测定系统来检测细胞毒素、TNF-α和TNF-β。