title: Biological containment of genetically modified Lactococcus lactis for intestinal delivery of human interleukin 10.
abstract: Genetically modified Lactococcus lactis secreting interleukin 10 provides a therapeutic approach for inflammatory bowel disease. However, the release of such genetically modified organisms through clinical use raises safety concerns. In an effort to address this problem, we replaced the thymidylate synthase gene thyA of L. lactis with a synthetic human IL10 gene. This thyA- hIL10+ L. lactis strain produced human IL-10 (hIL-10), and when deprived of thymidine or thymine, its viability dropped by several orders of magnitude, essentially preventing its accumulation in the environment. The biological containment system and the bacterium's capacity to secrete hIL-10 were validated in vivo in pigs. Our approach is a promising one for transgene containment because, in the unlikely event that the engineered L. lactis strain acquired an intact thyA gene from a donor such as L. lactis subsp. cremoris, the transgene would be eliminated from the genome.
authors.affiliation: Department of Molecular Biomedical Research, Vlaams Interuniversitair instituut voor Biotechnologie, Ghent University, KL. Ledeganckstraat 35, B-9000 Ghent, Belgium. l.steidler@ucc.ie
authors.forename: Lothar
authors.initials: L
authors.lastname: Steidler
authors.forename: Sabine
authors.initials: S
authors.lastname: Neirynck
authors.forename: Nathalie
authors.initials: N
authors.lastname: Huyghebaert
authors.forename: Veerle
authors.initials: V
authors.lastname: Snoeck
authors.forename: An
authors.initials: A
authors.lastname: Vermeire
authors.forename: Bruno
authors.initials: B
authors.lastname: Goddeeris
authors.forename: Eric
authors.initials: E
authors.lastname: Cox
authors.forename: Jean Paul
authors.initials: JP
authors.lastname: Remon
authors.forename: Erik
authors.initials: E
authors.lastname: Remaut
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002455:Cell Division; D002470:Cell Survival; D003092:Colitis / Q000382:microbiology / Q000628:therapy; D016503:Drug Delivery Systems / Q000379:methods; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology*; D005818:Genetic Engineering / Q000379:methods; D006801:Humans; D007082:Ileum / Q000382:microbiology*; D016753:Interleukin-10 / Q000235:genetics* / Q000378:metabolism* / Q000627:therapeutic use; D013294:Lactococcus lactis / Q000166:cytology / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D019936:Probiotics / Q000627:therapeutic use; D015202:Protein Engineering / Q000379:methods*; D013552:Swine; D013940:Thymidylate Synthase / Q000172:deficiency* / Q000235:genetics / Q000378:metabolism
chemical_list: D016753:Interleukin-10; D013940:Thymidylate Synthase