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Vpx-independent lentiviral transduction and shRNA-mediated protein knock-down in monocyte-derived dendritic cells

Wojciech Witkowski UGent, JOLIEN VERMEIRE, Alessia Landi, Evelien Naessens UGent, Hanne Vanderstraeten, Hans Nauwynck UGent, Herman Favoreel UGent and Bruno Verhasselt UGent (2015) PLOS ONE. 10(7).
abstract
The function of dendritic cells (DCs) in the immune system is based on their ability to sense and present foreign antigens. Powerful tools to research DC function and to apply in cell-based immunotherapy are either silencing or overexpression of genes achieved by lentiviral transduction. To date, efficient lentiviral transduction of DCs or their monocyte derived counterparts (MDDCs) required high multiplicity of infection (MOI) or the exposure to the HIV-2/SIV protein Vpx to degrade viral restriction factor SAM domain and HD domain-containing protein 1 (SAMHD1). Here we present a Vpx-independent method for efficient (>95%) transduction of MDDCs at lower MOI. The protocol can be used both for ectopic gene expression and knock-down. Introducing shRNA targeting viral entry receptor CD4 and restriction factor SAMHD1 into MDDCs resulted in down-regulation of targeted proteins and, consequently, expected impact on HIV infection. This protocol for MDDCs transduction is robust and free of the potential risk arising from the use of Vpx which creates a virus infection-prone environment, potentially dangerous in clinical setting.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
subject
keyword
MACROPHAGES, PATHOGENESIS, SAMHD1 RESTRICTS, HIV-1 RESTRICTION, IMMUNODEFICIENCY-VIRUS, REPLICATION, ACTIVATION, INFECTION, IMMUNITY
journal title
PLOS ONE
PLoS One
volume
10
issue
7
article number
e0133651
pages
12 pages
Web of Science type
Article
Web of Science id
000358622000115
JCR category
MULTIDISCIPLINARY SCIENCES
JCR impact factor
3.057 (2015)
JCR rank
11/63 (2015)
JCR quartile
1 (2015)
ISSN
1932-6203
DOI
10.1371/journal.pone.0133651
language
English
UGent publication?
yes
classification
A1
copyright statement
I have retained and own the full copyright for this publication
id
6960781
handle
http://hdl.handle.net/1854/LU-6960781
date created
2015-10-15 14:18:49
date last changed
2016-12-21 15:42:28
@article{6960781,
  abstract     = {The function of dendritic cells (DCs) in the immune system is based on their ability to sense and present foreign antigens. Powerful tools to research DC function and to apply in cell-based immunotherapy are either silencing or overexpression of genes achieved by lentiviral transduction. To date, efficient lentiviral transduction of DCs or their monocyte derived counterparts (MDDCs) required high multiplicity of infection (MOI) or the exposure to the HIV-2/SIV protein Vpx to degrade viral restriction factor SAM domain and HD domain-containing protein 1 (SAMHD1). Here we present a Vpx-independent method for efficient ({\textrangle}95\%) transduction of MDDCs at lower MOI. The protocol can be used both for ectopic gene expression and knock-down. Introducing shRNA targeting viral entry receptor CD4 and restriction factor SAMHD1 into MDDCs resulted in down-regulation of targeted proteins and, consequently, expected impact on HIV infection. This protocol for MDDCs transduction is robust and free of the potential risk arising from the use of Vpx which creates a virus infection-prone environment, potentially dangerous in clinical setting.},
  articleno    = {e0133651},
  author       = {Witkowski, Wojciech and VERMEIRE, JOLIEN and Landi, Alessia and Naessens, Evelien and Vanderstraeten, Hanne and Nauwynck, Hans and Favoreel, Herman and Verhasselt, Bruno},
  issn         = {1932-6203},
  journal      = {PLOS ONE},
  keyword      = {MACROPHAGES,PATHOGENESIS,SAMHD1 RESTRICTS,HIV-1 RESTRICTION,IMMUNODEFICIENCY-VIRUS,REPLICATION,ACTIVATION,INFECTION,IMMUNITY},
  language     = {eng},
  number       = {7},
  pages        = {12},
  title        = {Vpx-independent lentiviral transduction and shRNA-mediated protein knock-down in monocyte-derived dendritic cells},
  url          = {http://dx.doi.org/10.1371/journal.pone.0133651},
  volume       = {10},
  year         = {2015},
}

Chicago
Witkowski, Wojciech, Jolien Vermeire, Alessia Landi, Evelien Naessens, Hanne Vanderstraeten, Hans Nauwynck, Herman Favoreel, and Bruno Verhasselt. 2015. “Vpx-independent Lentiviral Transduction and shRNA-mediated Protein Knock-down in Monocyte-derived Dendritic Cells.” Plos One 10 (7).
APA
Witkowski, W., Vermeire, J., Landi, A., Naessens, E., Vanderstraeten, H., Nauwynck, H., Favoreel, H., et al. (2015). Vpx-independent lentiviral transduction and shRNA-mediated protein knock-down in monocyte-derived dendritic cells. PLOS ONE, 10(7).
Vancouver
1.
Witkowski W, Vermeire J, Landi A, Naessens E, Vanderstraeten H, Nauwynck H, et al. Vpx-independent lentiviral transduction and shRNA-mediated protein knock-down in monocyte-derived dendritic cells. PLOS ONE. 2015;10(7).
MLA
Witkowski, Wojciech, Jolien Vermeire, Alessia Landi, et al. “Vpx-independent Lentiviral Transduction and shRNA-mediated Protein Knock-down in Monocyte-derived Dendritic Cells.” PLOS ONE 10.7 (2015): n. pag. Print.