A peptidomimetic antagonist of TNF-alpha-mediated cytotoxicity identified from a phage-displayed random peptide library

• Published in: The Journal of immunology (1950) Vol. 161; no. 10; pp. 5621 - 5626
• Main Authors: Chirinos-Rojas, C L, Steward, M W, Partidos, C D
• Format: Journal Article
• Language: English
• Published: United States 15-11-1998
• Subjects: Amino Acid Sequence; Animals; Bacteriophages - immunology; Bacteriophages - isolation & purification; Bacteriophages - metabolism; Clone Cells; Cytotoxicity, Immunologic - drug effects; Humans; Mice; Molecular Mimicry; Molecular Sequence Data; Peptide Library; Peptides - immunology; Peptides - metabolism; Peptides - pharmacology; Protein Binding - immunology; Recombinant Proteins - metabolism; Sequence Analysis; Tumor Necrosis Factor-alpha - antagonists & inhibitors; Tumor Necrosis Factor-alpha - genetics; Tumor Necrosis Factor-alpha - metabolism
• ISSN: 0022-1767; 1550-6606
• DOI: 10.4049/jimmunol.161.10.5621

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.