title: Protection from diabetes development by single-chain antibody-mediated delivery of a NF-κB inhibitor specifically to β-cells in vivo.
abstract: Recently, we reported the generation of single-chain antibodies (SCAs) highly specific for rodent and human β-cells. Our current report describes the generation of a fusion protein of one of these SCAs (SCA B1) with a NF-κB essential modifier (NEMO)-binding domain (NBD) peptide, thereby creating a selective inhibitor of NF-κB activation in β-cells. The SCA B1-NBD fusion protein was cloned in the pIRES-EGFP, expressed in bacteria, and purified by metal affinity chromatography; the newly generated complex was then administered intravenously to rodents and evaluated for its ability to protect β-cells against cytokines in vitro and diabetogenic agents in vivo. First, it was shown clearly that our SCA B1-NBD fusion protein binds highly selective to CD rat β-cells in vivo. Second, we observed that SCA B1-mediated in vivo delivery of the NBD peptide completely blocked IL-1β + IFNγ- and TNFα + IFNγ-mediated induction of NF-κB as well as islet dysfunction in culture. Finally, repeated intravenous injection of SCA B1-NBD prior to multiple low-dose administration of streptozotocin in CD mice not only induced a striking resistance to diabetes development but also preserved β-cell mass. In conclusion, our data show for the first time that a SCA B1-NBD fusion peptide reliably protects β-cells against cytokines in vitro and allows protection from diabetes development in CD mice in vivo.
authors.affiliation: Department of Internal Medicine I, Division of Endocrinology and Metabolism, Berufsgenossenschaftliches University Hospital Bergmannsheil, Bochum, Germany.
authors.forename: Sandra
authors.initials: S
authors.lastname: Ueberberg
authors.forename: Timo
authors.initials: T
authors.lastname: Deutschbein
authors.forename: Harald H
authors.initials: HH
authors.lastname: Klein
authors.forename: Johannes W
authors.initials: JW
authors.lastname: Dietrich
authors.forename: Sara
authors.initials: S
authors.lastname: Akinturk
authors.forename: Nora
authors.initials: N
authors.lastname: Prochnow
authors.forename: Ralph
authors.initials: R
authors.lastname: Schirrmacher
authors.forename: Stephan
authors.initials: S
authors.lastname: Schneider
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D019610:Cytoprotection / Q000187:drug effects*; D003921:Diabetes Mellitus, Experimental / Q000139:chemically induced / Q000473:pathology / Q000517:prevention & control*; D016503:Drug Delivery Systems / Q000379:methods*; D004353:Drug Evaluation, Preclinical; D005260:Female; D050417:Insulin-Secreting Cells / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology / Q000502:physiology; D051379:Mice; D016328:NF-kappa B / Q000037:antagonists & inhibitors*; D009928:Organ Specificity / Q000187:drug effects; D010455:Peptides / Q000008:administration & dosage* / Q000494:pharmacology; D051381:Rats; D057127:Single-Chain Antibodies / Q000494:pharmacology*; D013311:Streptozocin; D013379:Substrate Specificity
chemical_list: C467063:NBD peptide, mouse; D016328:NF-kappa B; D010455:Peptides; D057127:Single-Chain Antibodies; D013311:Streptozocin