Protection from diabetes development by single-chain antibody-mediated delivery of a NF-κB inhibitor specifically to β-cells in vivo
| Autor: | Timo Deutschbein, Harald Klein, Ralph Schirrmacher, Johannes W. Dietrich, Sara Akinturk, Stephan Schneider, Sandra Ueberberg, Nora Prochnow |
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| Rok vydání: | 2011 |
| Předmět: |
medicine.medical_specialty
Physiology Endocrinology Diabetes and Metabolism medicine.medical_treatment Drug Evaluation Preclinical Streptozocin Diabetes Mellitus Experimental Substrate Specificity Mice chemistry.chemical_compound Drug Delivery Systems In vivo Insulin-Secreting Cells Physiology (medical) Diabetes mellitus Internal medicine medicine Animals Cells Cultured biology NF-kappa B NF-κB medicine.disease Streptozotocin Fusion protein Rats Cytokine Endocrinology chemistry Cytoprotection Organ Specificity biology.protein Female Antibody Beta cell Peptides Single-Chain Antibodies medicine.drug |
| Zdroj: | American Journal of Physiology-Endocrinology and Metabolism. 301:E83-E90 |
| ISSN: | 1522-1555 0193-1849 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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