Human interleukin-23 receptor antagonists derived from an albumin-binding domain scaffold inhibit IL-23-dependent ex vivo expansion of IL-17-producing T-cells

0431238 - BTÚ 2015 RIV US eng J - Článek v odborném periodiku
Kuchař, Milan - Vaňková, Lucie - Petroková, Hana - Černý, Jiří - Osička, Radim - Pelák, O. - Šípová, Hana - Schneider, Bohdan - Homola, Jiří - Šebo, Peter - Kalina, T. - Malý, Petr
Human interleukin-23 receptor antagonists derived from an albumin-binding domain scaffold inhibit IL-23-dependent ex vivo expansion of IL-17-producing T-cells.
Proteins-Structure, Function and Bioinformatics. Roč. 82, č. 6 (2014), s. 975-989. ISSN 0887-3585. E-ISSN 1097-0134
Grant CEP: GA ČR GAP303/10/1849; GA ČR(CZ) GAP302/11/0580; GA MŠMT(CZ) ED1.1.00/02.0109
Výzkumný záměr: CEZ:AV0Z50520701
Institucionální podpora: RVO:86652036 ; RVO:61388971 ; RVO:67985882
Klíčová slova: ribosome display * combinatorial library * psoriasis
Kód oboru RIV: EB - Genetika a molekulární biologie
Impakt faktor: 2.627, rok: 2014

Engineered combinatorial libraries derived from small protein scaffolds represent a powerful tool for generating novel binders with high affinity, required specificity and designed inhibitory function. This work was aimed to generate a collection of recombinant binders of human interleukin-23 receptor (IL-23R), which is a key element of proinflammatory IL-23-mediated signaling. A library of variants derived from the three-helix bundle scaffold of the albumin-binding domain (ABD) of streptococcal protein G and ribosome display were used to select for high-affinity binders of recombinant extracellular IL-23R. A collection of 34 IL-23R-binding proteins (called REX binders), corresponding to 18 different sequence variants, was used to identify a group of ligands that inhibited binding of the recombinant p19 subunit of IL-23, or the biologically active human IL-23 cytokine, to the recombinant IL-23R or soluble IL-23R-IgG chimera. The strongest competitors for IL-23R binding in ELISA were confirmed to recognize human IL-23R-IgG in surface plasmon resonance experiments, estimating the binding affinity in the sub- to nanomolar range. We further demonstrated that several REX variants bind to human leukemic cell lines K-562, THP-1 and Jurkat, and this binding correlated with IL-23R cell-surface expression. The REX125, REX009 and REX128 variants competed with the p19 protein for binding to THP-1 cells. Moreover, the presence of REX125, REX009 and REX115 variants significantly inhibited the IL-23-driven expansion of IL-17-producing primary human CD4(+) T-cells. Thus, we conclude that unique IL-23R antagonists derived from the ABD scaffold were generated that might be useful in designing novel anti-inflammatory biologicals. Proteins 2014; 82:975-989. (c) 2013 The Authors. Proteins: Structure, Function, and Bioinformatics Published by Wiley Periodicals, Inc.
Trvalý link: http://hdl.handle.net/11104/0235830