TNF－α的生物活性检测法与酶联免疫吸附法比较

Comparison of Bioassay and Enzyme-Linked Immunosorbent Assay for Measurement of TNF-α

比较了生物活性检测法（ｂｉｏａｓｓｙ，ＢＡ）与酶联免疫吸附法（ＥＬＩＳＡ）对大鼠腹腔巨噬细胞（ｍａｃｒｏｐｈａｇｅｓ，Ｍφ）分泌的肿瘤坏死因子－α（ｔｕｍｏｒｎｅｃｒｏｓｉｓｆａｃｔｏｒ－α，ＴＮＦ－α）的检测。在静息Ｍφ培养上清液中，用两种方法均未测到ＴＮＦ－α；而脂多糖（ｌｉｐｏｐｏｌｙｓａｃｃｈａｒｉｄｅ，ＬＰＳ，１００μｇ／Ｌ）刺激组，用两种方法均测到大量ＴＮＦ－α；如ＬＰＳ与秋水仙碱（ｃｏｌｃｈｉｃｉｎｅ，Ｃｏｌ）联用，ＴＮＦ－α的产生受到抑制，ＢＡ法检测显示抑制率为９７％，而ＥＬＩＳＡ检测则显示抑制率为４２％，提示Ｃｏｌ对ＴＮＦ－α生物活性的抑制比对其生物合成的抑制更明显。此外，Ｃｏｌ在浓度≥１０μｍｏｌ／Ｌ时，可部分减轻ＴＮＦ－α对其靶细胞Ｌ９２９的毒性作用。文中对这两种方法的优缺点进行了比较与讨论。

The results of bioassay and ELISA in the test of TNF-a release from rat peritoneal macrophage(Mφ)were compared.In the supernatant of resident Mφ culture no measurable TNF-α was found by both methods. In the Lipopolysaccharide(LPS)(100μg/L)stimulated group,however,markedly enhanced TNF-α could be demonstrated by both assays. When Mφs were treated with LPS and colchicine(Col),it has been shown that TNF-α production was suppressed.The inhibitory rate of LPS-induced TNF-α by Col was 97% by using bioassay, but only 42%by using ELISA,This suggested that the suppressive effect of Col on TNF-α bioactivity is more significant than that on TNF-α biosynthesis.Furthermore,the TNF-α toxicity to its target cell-L929 was also decreased by Col treatment(≥10 μmol/L).The advantages and disadvantages between the two methods were discussed.