title: Intracellular accumulation of trehalose protects Lactococcus lactis from freeze-drying damage and bile toxicity and increases gastric acid resistance.
abstract: Interleukin-10 (IL-10) is a promising candidate for the treatment of inflammatory bowel disease. Intragastric administration of Lactococcus lactis genetically modified to secrete IL-10 in situ in the intestine was shown to be effective in healing and preventing chronic colitis in mice. However, its use in humans is hindered by the sensitivity of L. lactis to freeze-drying and its poor survival in the gastrointestinal tract. We expressed the trehalose synthesizing genes from Escherichia coli under control of the nisin-inducible promoter in L. lactis. Induced cells accumulated intracellular trehalose and retained nearly 100% viability after freeze-drying, together with a markedly prolonged shelf life. Remarkably, cells producing trehalose were resistant to bile, and their viability in human gastric juice was enhanced. None of these effects were seen with exogenously added trehalose. Trehalose accumulation did not interfere with IL-10 secretion or with therapeutic efficacy in murine colitis. The newly acquired properties should enable a larger proportion of the administered bacteria to reach the gastrointestinal tract in a bioactive form, providing a means for more effective mucosal delivery of therapeutics.
authors.affiliation: Department for Molecular Biomedical Research, Flanders Interuniversity Institute for Biotechnology (VIB) and Ghent University, Technologiepark 927, B-9052 Ghent, Belgium.
authors.forename: Sofie
authors.initials: S
authors.lastname: Termont
authors.forename: Klaas
authors.initials: K
authors.lastname: Vandenbroucke
authors.forename: Dirk
authors.initials: D
authors.lastname: Iserentant
authors.forename: Sabine
authors.initials: S
authors.lastname: Neirynck
authors.forename: Lothar
authors.initials: L
authors.lastname: Steidler
authors.forename: Erik
authors.initials: E
authors.lastname: Remaut
authors.forename: Pieter
authors.initials: P
authors.lastname: Rottiers
mesh_terms: D000818:Animals; D001646:Bile / Q000737:chemistry; D002908:Chronic Disease; D003092:Colitis / Q000382:microbiology / Q000628:therapy*; D003451:Cryoprotective Agents / Q000378:metabolism / Q000494:pharmacology; D016503:Drug Delivery Systems; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D005260:Female; D005612:Freeze Drying; D005750:Gastric Juice / Q000737:chemistry; D041981:Gastrointestinal Tract / Q000382:microbiology*; D005964:Glucosyltransferases / Q000235:genetics / Q000378:metabolism; D006801:Humans; D016753:Interleukin-10 / Q000235:genetics / Q000378:metabolism; D013294:Lactococcus lactis / Q000187:drug effects / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014199:Trehalose / Q000378:metabolism* / Q000494:pharmacology
chemical_list: D003451:Cryoprotective Agents; D029968:Escherichia coli Proteins; D011994:Recombinant Proteins; D016753:Interleukin-10; D014199:Trehalose; D005964:Glucosyltransferases; C023180:trehalose-6-phosphate synthase