Journal of Transmitted Diseases and Immunity

ER - intrabody mediates knockdown of mouse IFN alpha in macrophages and dendritic cells

Joint Event on 22^nd Edition of International Conference on Immunology and Evolution of Infectious Diseases & 12^th Edition of International Conference on Tissue Engineering and Regenerative Medicine
May 10-11, 2018 Frankfurt, Germany

Thomas Boldicke

Helmholtz Centre for Infection Research, Germany

ScientificTracks Abstracts: J Transm Dis Immun

DOI: 10.21767/2573-0320-C2-005

Abstract

IFN-�?± activates the transcription of various IFN-stimulated genes (ISGs) in virus infected cells. Proteins encoded by ISGs block viral transport into the host cell and inhibit viral gene transcription and translation. Due to the existence of 13 different high homologous isoforms of mouse IFN-�?±, an IFN-�?± knockout mouse has not yet been established by conventional knockout strategies and CRISPR/Cas. We used an IFN-�?± knockdown strategy based on ER-intrabodies to inhibit IFN-�?± secretion in macrophages and dendritic cells, the main producers of IFN-�?±ï¢ï¢after virus infection. To realize this strategy an ER intrabody was constructed from an anti-mouse IFN-�?± rat hybridoma recognizing five mice IFN-�?± isoforms. We follow the hypothesis that an intrabody recognizing five high homologous isoforms of the proteins will be able to knockdown all isoforms. The secretion of IFN-�?± was significantly inhibited by the intrabody in stable intrabody expressing RAW 264.7 macrophages and D1 dendritic cells as demonstrated by LISA, Mx2-dependent luciferase assay and immunofluorescence. This antibody has the potential to knockdown IFN-�?± in transgenic intrabody mice. These animals might be very valuable in the future to study in detail the role of IFN-�?± during active and chronic viral infections and in autoimmune diseases.

Biography

Thomas Boldicke has received his PhD in 1982 at the Max-Planck-Institut of Molecular Genetics, Berlin. He started his carrier as Postdoc at the German Research Centre for Biotechnology (GBF, Brunswick) in the Department of Genetics and Cell Biology by John Collins. Now he is a Senior Scientist at the Helmholtz Centre for Infection Research (HZI, former GBF) and Project Leader of intrabodies. In 2011, he qualified as a Professor in Molecular Biology and Cell Biology at the Technical University of Braunschweig. He is an expert in generating mouse and human hybridomas and in selecting and modifying recombinant antibodies. In the last decade he focused on the construction and characterization of intracellular antibodies. He has published 35 manuscripts.

Email:thomas.boeldicke@helmholtz hzi.de