title: Improving health from the inside: Use of engineered intestinal microorganisms as in situ cytokine delivery system.
abstract: The anti-inflammatory cytokine interleukin-10 and its viral homologs were chosen as model proteins for the development of drug delivery systems based on probiotic carriers like E. coli Nissle 1917, E. coli G3/10, and Saccharomyces boulardii. Exterior cytokine secretion was achieved by a modified E. coli hemolysin transporter. Release of interleukin-10 transported to the periplasm via the OmpF signal peptide was enabled by a T4 phage lysis system under control of the araC PBAD activator-promoter. The yield of interleukin-10 delivered by the phage lysis system was too low for functional analysis whereas the fusion protein secreted by the hemolysin transporter proved to be biologically inactive. Moreover, partial processing of the fusion protein by the E. coli membrane protease OmpT had no effect on the protein's functionality. Using the α-mating factor signal sequence, the yeast S. boulardii proved to be suitable for secretory expression of biologically active viral interleukin-10.
authors.affiliation: Department of Laboratory Medicine, Robert-Bosch Hospital, Stuttgart, Germany. Christoph.Poehlmann@rbk.de
authors.forename: Christoph
authors.initials: C
authors.lastname: Pöhlmann
authors.forename: Mandy
authors.initials: M
authors.lastname: Thomas
authors.forename: Sarah
authors.initials: S
authors.lastname: Förster
authors.forename: Manuela
authors.initials: M
authors.lastname: Brandt
authors.forename: Maike
authors.initials: M
authors.lastname: Hartmann
authors.forename: André
authors.initials: A
authors.lastname: Bleich
authors.forename: Florian
authors.initials: F
authors.lastname: Gunzer
mesh_terms: D000818:Animals; D002460:Cell Line; D016503:Drug Delivery Systems / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D005818:Genetic Engineering; D006460:Hemolysin Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D016753:Interleukin-10 / Q000235:genetics* / Q000378:metabolism; D007422:Intestines / Q000382:microbiology*; D051379:Mice; D019936:Probiotics / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012440:Saccharomyces / Q000235:genetics* / Q000378:metabolism
chemical_list: D029968:Escherichia coli Proteins; D006460:Hemolysin Proteins; C036507:Hlya protein, E coli; D011993:Recombinant Fusion Proteins; D016753:Interleukin-10
keywords: Escherichia coli; Saccharomyces boulardii; bacterial cell lysis; biological containment; hemolysin A; interleukin-10; probiotics; transport system