Chirinos-Rojas C L, Steward M W, Partidos C D
Department of Infections and Tropical Diseases, London School of Hygiene and Tropical Medicine, United Kingdom.
J Immunol. 1998 Nov 15;161(10):5621-6.
Phage-displayed peptide libraries represent a vast collection of peptide sequences that can be used to identify novel therapeutic molecules. In this report, a 15-mer phage-displayed peptide library was used to identify potential TNF-alpha antagonists. After direct interaction of recombinant human TNF-alpha with the library, four randomly selected phage clones were shown to inhibit in a dose-dependent fashion both mouse and human TNF-alpha-induced cytotoxicity in vitro. DNA sequencing of the positive clones revealed a common amino acid sequence that does not bear any structural similarity to the known primary structures of the extracellular domains of either 55-kDa or 75-kDa TNF receptors. This sequence was synthesized, and the peptidomimotope was shown i) to bind to the recombinant human TNF-alpha using surface plasmon resonance (biosensor) technology and ii) to inhibit both recombinant mouse and human TNF-alpha-induced cytotoxicity in vitro in a dose-dependent fashion. These findings highlight the potential of phage-displayed random peptide libraries for the identification of novel low molecular antagonistic molecules that can block the biologic activities of TNF-alpha.
噬菌体展示肽库代表了大量的肽序列集合，可用于鉴定新型治疗分子。在本报告中，一个15肽的噬菌体展示肽库被用于鉴定潜在的肿瘤坏死因子-α（TNF-α）拮抗剂。重组人TNF-α与该肽库直接相互作用后，四个随机选择的噬菌体克隆在体外以剂量依赖的方式抑制了小鼠和人TNF-α诱导的细胞毒性。对阳性克隆进行DNA测序，发现了一个共同的氨基酸序列，该序列与55 kDa或75 kDa TNF受体细胞外结构域的已知一级结构没有任何结构相似性。合成了该序列，并且该肽模拟物被证明：i）使用表面等离子体共振（生物传感器）技术与重组人TNF-α结合；ii）在体外以剂量依赖的方式抑制重组小鼠和人TNF-α诱导的细胞毒性。这些发现突出了噬菌体展示随机肽库在鉴定可阻断TNF-α生物学活性的新型低分子拮抗剂方面的潜力。
