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Journal of Transmitted Diseases and Immunity
ISSN: 2573-0320
Page 50
Volume 4
May 10-11, 2018
Frankfurt, Germany
Immunology Research 2018
Tissue Science 2018
JOINT EVENT
2 2
n d
E d i t i o n o f I n t e r n a t i o n a l C o n f e r e n c e o n
Immunology and
Evolution of Infectious Diseases
&
1 2
t h
E d i t i o n o f I n t e r n a t i o n a l C o n f e r e n c e o n
Tissue Engineering and
Regenerative Medicine
I
FN-
α
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-
α
, an IFN-
α
knockout mouse
has not yet been established by conventional knockout strategies
and CRISPR/Cas. We used an IFN-
α
knockdown strategy based
on ER-intrabodies to inhibit IFN-
α
secretion in macrophages and
dendritic cells, the main producers of IFN-
α
after virus infection.
To realize this strategy an ER intrabody was constructed from
an anti-mouse IFN-
α
rat hybridoma recognizing five mice IFN-
α
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-
α
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-
α
in transgenic
intrabody mice. These animals might be very valuable in the
future to study in detail the role of IFN-
α
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 Biol-
ogy 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 pub-
lished 35 manuscripts.
thomas.boeldicke@helmholtz
hzi.deER - intrabody mediates knockdown of mouse IFN alpha
in macrophages and dendritic cells
Thomas Boldicke
Helmholtz Centre for Infection Research, Germany
Thomas Boldicke, J Transm Dis Immun 2018, Volume 2
DOI: 10.21767/2573-0320-C2-005