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p19-Targeting ILP Protein Blockers of IL-23/Th-17 Pro-Inflammatory Axis Displayed on Engineered Bacteria of Food Origin
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SYSNO ASEP 0498920 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title p19-Targeting ILP Protein Blockers of IL-23/Th-17 Pro-Inflammatory Axis Displayed on Engineered Bacteria of Food Origin Author(s) Skrlec, K. (SI)
Zadravec, P. (SI)
Hlavničková, Marie (BTO-N)
Kuchař, Milan (BTO-N) RID
Vaňková, Lucie (BTO-N)
Petroková, Hana (BTO-N) RID
Křížová, Lucie (BTO-N)
Černý, Jiří (BTO-N) RID, ORCID
Berlec, A. (SI)
Malý, Petr (BTO-N) RID, ORCIDNumber of authors 10 Article number 1933 Source Title International Journal of Molecular Sciences. - : MDPI
Roč. 19, č. 7 (2018)Number of pages 14 s. Language eng - English Country CH - Switzerland Keywords lactococcus ; binding protein ; albumin-binding domain Subject RIV EB - Genetics ; Molecular Biology OECD category Biochemistry and molecular biology R&D Projects NV16-27676A GA MZd - Ministry of Health (MZ) ED1.1.00/02.0109 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) Institutional support BTO-N - RVO:86652036 UT WOS 000442807400112 EID SCOPUS 85049569279 DOI 10.3390/ijms19071933 Annotation IL-23-mediated Th-17 cell activation and stimulation of IL-17-driven pro-inflammatory axis has been associated with autoimmunity disorders such as Inflammatory Bowel Disease (IBD) or Crohn's Disease (CD). Recently we developed a unique class of IL-23-specific protein blockers, called ILP binding proteins that inhibit binding of IL-23 to its cognate cell-surface receptor (IL-23R) and exhibit immunosuppressive effect on human primary blood leukocytes ex vivo. In this study, we aimed to generate a recombinant Lactococcus lactis strain which could serve as in vivo producer/secretor of IL-23 protein blockers into the gut. To achieve this goal, we introduced ILP030, ILP317 and ILP323 cDNA sequences into expression plasmid vector containing USP45 secretion signal, FLAG sequence consensus and LysM-containing cA surface anchor (AcmA) ensuring cell-surface peptidoglycan anchoring. We demonstrate that all ILP variants are expressed in L. lactis cells, efficiently transported and secreted from the cell and displayed on the bacterial surface. The binding function of AcmA-immobilized ILP proteins is documented by interaction with a recombinant p19 protein, alpha subunit of human IL-23, which was assembled in the form of a fusion with Thioredoxin A. ILP317 variant exhibits the best binding to the human IL-23 cytokine, as demonstrated for particular L. lactis-ILP recombinant variants by Enzyme-Linked ImmunoSorbent Assay (ELISA). We conclude that novel recombinant ILP-secreting L. lactis strains were developed that might be useful for further in vivo studies of IL-23-mediated inflammation on animal model of experimentally-induced colitis. Workplace Institute of Biotechnology Contact Monika Kopřivová, Monika.Koprivova@ibt.cas.cz, Tel.: 325 873 700 Year of Publishing 2019
Number of the records: 1