OF A SOLID-PHASE RANDOM PEPTIDE LIBRARY TO IDENTIFY INHIBITORS OF TNF-ALPHA MEDIATED CYTOTOXICITY IN-VITRO

Tumour necrosis factor (TNF) is a pleiotropic cytokine which plays a c entral role in infection, inflammation and autoimmune diseases, Its fu nctions are mediated through binding to high affinity cell surface rec eptors, An approach to modulate excessive levels of TNF-alpha in the s erum is the use of soluble receptors, In this study the potential of a solid-phase combinatorial peptide library was investigated to identif y peptide mimics of the binding site of the TNF-alpha receptor, One of the identified mimotopes was shown to inhibit TNF-alpha-mediated cyto toxicity in mouse L929 and in a human KYM-1D4 cell lines in a dose-dep endent fashion, Characterization of the mimotope sequence by high-perf ormance liquid chromatography and mass spectroscopy has shown that the inhibitory effect of the mimotope was dependent upon the presence of a protective MTR group on the side chains of the arginine residues in the mimotope sequence, Furthermore, antibodies to the mimotope were sh own to recognize the remombinant human TNF 55-kDa receptor in an ELISA assay, These findings highlight the potential of combinatorial peptid e libraries in identifying peptide mimics of the binding site of TNF-a lpha receptor, which can be used as competive inhibitors of ligand-rec eptor interactions. (C) 1997 Academic Press Limited.