Gouze Jean-Noel, Gouze Elvire, Palmer Glyn D, Liew Victor S, Pascher Arnulf, Betz Oliver B, Thornhill Thomas S, Evans Christopher H, Grodzinsky Alan J, Ghivizzani Steven C
Center for Molecular Orthopaedics, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, USA.
Arthritis Res Ther. 2003;5(5):R301-9. doi: 10.1186/ar795. Epub 2003 Aug 1.
Anakinra, the recombinant form of IL-1 receptor antagonist (IL-1Ra), has been approved for clinical use in the treatment of rheumatoid arthritis as the drug Kineret trade mark, but it must be administered daily by subcutaneous injection. Gene transfer may offer a more effective means of delivery. In this study, using prostaglandin E2 production as a measure of stimulation, we quantitatively compared the ability of anakinra, as well as that of IL-1Ra delivered by gene transfer, to inhibit the biologic actions of IL-1beta. Human synovial fibroblast cultures were incubated with a range of doses of anakinra or HIG-82 cells genetically modified to constitutively express IL-1Ra. The cultures were then challenged with recombinant human IL-1beta either simultaneously with addition of the source of IL-1Ra or 24 hours later. In a similar manner, the potencies of the two sources of IL-1Ra were compared when human synovial fibroblasts were challenged with IL-1beta produced constitutively by genetically modified cells. No significant difference in inhibitory activity was observed between recombinant protein and IL-1Ra provided by the genetically modified cells, under static culture conditions, even following incubation for 4 days. However, under culture conditions that provided progressive dilution of the culture media, striking differences between these methods of protein delivery became readily apparent. Constitutive synthesis of IL-1Ra by the genetically modified cells provided sustained or increased protection from IL-1 stimulation over time, whereas the recombinant protein became progressively less effective. This was particularly evident under conditions of continuous IL-1beta synthesis.
阿那白滞素是白细胞介素-1受体拮抗剂(IL-1Ra)的重组形式,已作为Kineret商标药物被批准用于类风湿性关节炎的临床治疗,但必须每天皮下注射给药。基因转移可能提供一种更有效的给药方式。在本研究中,以前列腺素E2的产生作为刺激的衡量指标,我们定量比较了阿那白滞素以及通过基因转移递送的IL-1Ra抑制IL-1β生物学作用的能力。将人滑膜成纤维细胞培养物与一系列剂量的阿那白滞素或经基因改造以组成性表达IL-1Ra的HIG-82细胞一起孵育。然后在添加IL-1Ra来源的同时或24小时后,用重组人IL-1β刺激培养物。同样地,当用人滑膜成纤维细胞受到经基因改造的细胞组成性产生的IL-1β刺激时,比较了两种IL-1Ra来源的效力。在静态培养条件下,即使孵育4天后,重组蛋白和经基因改造的细胞提供的IL-1Ra之间在抑制活性上也未观察到显著差异。然而,在提供培养基逐步稀释的培养条件下,这些蛋白质递送方法之间的显著差异变得很明显。经基因改造的细胞组成性合成IL-1Ra随着时间的推移提供了对IL-1刺激的持续或增强的保护,而重组蛋白的效果则逐渐降低。在持续合成IL-1β的条件下尤其明显。