Evans T J, Moyes D, Carpenter A, Martin R, Loetscher H, Lesslauer W, Cohen J
Department of Infectious Diseases and Bacteriology, Royal Postgraduate Medical School, London, UK.
J Exp Med. 1994 Dec 1;180(6):2173-9. doi: 10.1084/jem.180.6.2173.
The aim of this study was to compare the ability of both a 55- and 75-kD soluble tumor necrosis factor receptor immunoglobulin G fusion protein (sTNFR-IgG) in protecting against death in a murine model of gram-negative sepsis. Pretreatment with 250 micrograms of the p75 construct delayed but did not avert death in this model, reducing peak bioactive TNF-alpha levels after infection from 76.4 ng ml-1 in control mice to 4.7 ng ml-1 in the treated group (p < 0.05, two-sample t test). However, these low levels of bioactive TNF-alpha persisted in the p75 fusion protein-treated animals compared with the controls and were sufficient to mediate delayed death. In contrast, pretreatment with 200 micrograms of the p55 sTNFR-IgG gave excellent protection against death with complete neutralization of circulating TNF. Studies of the binding of TNF-alpha with the soluble TNFR fusion proteins showed that the p75 fusion construct exchanges bound TNF-alpha about 50-100-fold faster than the p55 fusion protein. Thus, although both fusion proteins in equilibrium bind TNF-alpha with high affinity, the TNF-alpha p55 fusion protein complex is kinetically more stable than the p75 fusion construct, which thus acts as a TNF carrier. The persistent release of TNF-alpha from the p75 fusion construct limits its therapeutic effect in this model of sepsis.
本研究的目的是比较55-kD和75-kD可溶性肿瘤坏死因子受体免疫球蛋白G融合蛋白(sTNFR-IgG)在革兰氏阴性菌败血症小鼠模型中预防死亡的能力。在该模型中,用250微克p75构建体进行预处理可延迟但不能避免死亡,感染后生物活性TNF-α的峰值水平从对照小鼠的76.4 ng/ml降至治疗组的4.7 ng/ml(p<0.05,双样本t检验)。然而,与对照组相比,p75融合蛋白处理的动物中这些低水平的生物活性TNF-α持续存在,并且足以介导延迟死亡。相比之下,用200微克p55 sTNFR-IgG进行预处理可提供极佳的死亡保护,使循环中的TNF完全中和。对TNF-α与可溶性TNFR融合蛋白结合的研究表明,p75融合构建体交换结合的TNF-α的速度比p55融合蛋白快约50-100倍。因此,尽管两种融合蛋白在平衡状态下都以高亲和力结合TNF-α,但TNF-α p55融合蛋白复合物在动力学上比p75融合构建体更稳定,因此p75融合构建体起到了TNF载体的作用。在该败血症模型中,p75融合构建体持续释放TNF-α限制了其治疗效果。