Počet záznamů: 1  

Engineered Lactococcus lactis Secreting IL-23 Receptor-Targeted REX Protein Blockers for Modulation of IL-23/Th17-Mediated Inflammation

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    SYSNO ASEP0507592
    Druh ASEPJ - Článek v odborném periodiku
    Zařazení RIVJ - Článek v odborném periodiku
    Poddruh JČlánek ve WOS
    NázevEngineered Lactococcus lactis Secreting IL-23 Receptor-Targeted REX Protein Blockers for Modulation of IL-23/Th17-Mediated Inflammation
    Tvůrce(i) Plavec, T. V. (SI)
    Kuchař, Milan (BTO-N) RID
    Benko, A. (SI)
    Lišková, Veronika (BTO-N)
    Černý, Jiří (BTO-N) RID, ORCID
    Berlec, A. (SI)
    Malý, Petr (BTO-N) RID, ORCID
    Celkový počet autorů7
    Číslo článku152
    Zdroj.dok.Microorganisms. - : MDPI
    Roč. 7, č. 5 (2019)
    Poč.str.13 s.
    Jazyk dok.eng - angličtina
    Země vyd.CH - Švýcarsko
    Klíč. slovalactococcus ; binding protein ; albumin-binding domain ; IL-23 cytokine
    Vědní obor RIVEE - Mikrobiologie, virologie
    Obor OECDMicrobiology
    CEPNV16-27676A GA MZd - Ministerstvo zdravotnictví
    ED1.1.00/02.0109 GA MŠk - Ministerstvo školství, mládeže a tělovýchovy
    Způsob publikováníOpen access
    Institucionální podporaBTO-N - RVO:86652036
    UT WOS000470962000040
    EID SCOPUS85074238090
    DOI10.3390/microorganisms7050152
    AnotaceLactococcus lactis, a probiotic bacterium of food origin, has recently been demonstrated as a suitable strain for the production and in vivo delivery of therapeutically important proteins into the gut. We aimed to engineer recombinant L. lactis cells producing/secreting REX binding proteins that have been described as IL-23 receptor (IL-23R) blockers and IL-23R antagonists suppressing the secretion of cytokine IL-17A, a pivotal step in the T-helper Th17-mediated pro-inflammatory cascade, as well as in the development of autoimmune diseases, including inflammatory bowel disease (IBD). To reach this goal, we introduced cDNA sequences coding for REX009, REX115, and REX125 proteins into plasmid vectors carrying a Usp45 secretion signal, a FLAG tag sequence consensus, and a LysM-containing cA surface anchor (AcmA), thus allowing cell-surface peptidoglycan anchoring. These plasmids, or their non-FLAG/non-AcmA versions, were introduced into L. lactis host cells, thus generating unique recombinant L. lactis-REX strains. We demonstrate that all three REX proteins are expressed in L. lactis cells and are efficiently displayed on the bacterial surface, as tested by flow cytometry using an anti-FLAG antibody conjugate. Upon 10-fold concentration of the conditioned media, a REX125 secretory variant can be detected by Western blotting. To confirm that the FLAG/non-FLAG REX proteins displayed by L. lactis retain their binding specificity, cell-surface interactions of REX proteins with an IL-23R-IgG chimera were demonstrated by flow cytometry. In addition, statistically significant binding of secreted REX009 and REX115 proteins to bacterially produced, soluble human IL-23R was confirmed by ELISA. We conclude that REX-secreting L. lactis strains were engineered that might serve as IL-23/IL-23R blockers in an experimentally induced mouse model of colitis.
    PracovištěBiotechnologický ústav
    KontaktMonika Kopřivová, Monika.Koprivova@ibt.cas.cz, Tel.: 325 873 700
    Rok sběru2020
    Elektronická adresahttps://www.mdpi.com/2076-2607/7/5/152
Počet záznamů: 1