Campbell G R, Magee D, Kennedy A, Rowlands B J, Halliday M I
Department of Surgery, Queen's University of Belfast, Northern Ireland.
Eur Cytokine Netw. 1997 Mar;8(1):97-102.
Tumour necrosis factor (TNF) is a cytokine with multiple biological activities which plays a pivotal role in the response of the body to infection. TNF is secreted in the the monomeric form and associates to yield a biologically active oligomeric molecule. Bioactive TNF can be measured in plasma using cytotoxic assays which employ the murine cell-lines L929 or WEHI 164 clone 13. However, it has become clear that inactive TNF also circulates in vivo, which is not detected by bioassay. These inactive forms may play an important role in the regulation of TNF activity. The rat is often used as a model for the study of acute infection and its systemic effects. Our aim was to develop an ELISA for rat TNF which would provide a convenient and cost effective method of assay. The assay employs two commercially available antibodies raised against recombinant murine TNF (rMuTNF) which exhibit cross-reactivity with rat TNF. The lower limit of detection for this assay was determined to be 39.0 pg/ml rMuTNF. The inter and intra-assay coefficients of variation were < 12.0%. Specificity of the assay was shown by the high degree of parallelism obtained between rMuTNF and commercially available rRatTNF. The assay described measures rat TNF in both plasma and tissue culture supernatant. The measurement of plasma concentrations of TNF using both the ELISA and bioassay may help elucidate more fully the biological importance of TNF.
肿瘤坏死因子（TNF）是一种具有多种生物学活性的细胞因子，在机体对感染的反应中起关键作用。TNF以单体形式分泌，并结合形成具有生物活性的寡聚分子。可使用采用小鼠细胞系L929或WEHI 164克隆13的细胞毒性试验在血浆中检测生物活性TNF。然而，现已明确无活性的TNF也在体内循环，生物测定法无法检测到。这些无活性形式可能在TNF活性调节中起重要作用。大鼠常被用作研究急性感染及其全身效应的模型。我们的目的是开发一种用于大鼠TNF的酶联免疫吸附测定（ELISA）方法，该方法将提供一种方便且经济高效的检测方法。该测定采用两种针对重组小鼠TNF（rMuTNF）产生的市售抗体，它们与大鼠TNF具有交叉反应性。该测定的检测下限确定为39.0 pg/ml rMuTNF。测定的批间和批内变异系数均<12.0%。rMuTNF与市售rRatTNF之间获得的高度平行性表明了该测定的特异性。所描述的测定法可测量血浆和组织培养上清液中的大鼠TNF。使用ELISA和生物测定法测量血浆中TNF的浓度可能有助于更全面地阐明TNF的生物学重要性。
