Visualization of DC-SIGN-mediated entry pathway of engineered lentiviral vectors in target cells.
Dendritic cells (DCs) are potent antigen-presenting cells and therefore have enormous potential as vaccine targets. We have previously developed an engineered lentiviral vector (LV) that is pseudotyped with a mutated Sindbis virus glycoprotein (SVGmu), which is capable of targeting DCs through Dendr...
Main Authors: | , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
Public Library of Science (PLoS)
2013-01-01
|
Series: | PLoS ONE |
Online Access: | http://europepmc.org/articles/PMC3696072?pdf=render |
id |
doaj-a2e3bb3e79984324b33f696a948c868c |
---|---|
record_format |
Article |
spelling |
doaj-a2e3bb3e79984324b33f696a948c868c2020-11-25T00:26:49ZengPublic Library of Science (PLoS)PLoS ONE1932-62032013-01-0186e6740010.1371/journal.pone.0067400Visualization of DC-SIGN-mediated entry pathway of engineered lentiviral vectors in target cells.Yarong LiuApril TaiKye-Il JooPin WangDendritic cells (DCs) are potent antigen-presenting cells and therefore have enormous potential as vaccine targets. We have previously developed an engineered lentiviral vector (LV) that is pseudotyped with a mutated Sindbis virus glycoprotein (SVGmu), which is capable of targeting DCs through Dendritic Cell-specific ICAM3-grabbing Nonintegrin (DC-SIGN), a receptor that is predominantly expressed by DCs. In this study, we aimed to elucidate the internalization and trafficking mechanisms of this viral vector system through direct visualization of GFP-Vpr-tagged viral particles in target DCs, which was further corroborated by drug inhibition and dominant-negative mutants of cellular proteins that regulate the endocytic traffic. We demonstrated that our engineered LVs enter the cell via receptor-mediated clathrin- and dynamin-dependent endocytosis. Microtubule networks were also involved in a productive infection. Viral vector fusion was low-pH-dependent and occurred in the early endosomal stage of the intracellular transport. Autophagy was also examined for its effect on transduction efficiency, and we observed that enhanced autophage activity reduced vector infectivity, while suppressed autophagy boosted transduction efficiency. This study shed some light on the internalization and trafficking mechanisms of DC-directed LVs and offers some strategies to further improve the efficiency of LV-mediated gene therapy.http://europepmc.org/articles/PMC3696072?pdf=render |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Yarong Liu April Tai Kye-Il Joo Pin Wang |
spellingShingle |
Yarong Liu April Tai Kye-Il Joo Pin Wang Visualization of DC-SIGN-mediated entry pathway of engineered lentiviral vectors in target cells. PLoS ONE |
author_facet |
Yarong Liu April Tai Kye-Il Joo Pin Wang |
author_sort |
Yarong Liu |
title |
Visualization of DC-SIGN-mediated entry pathway of engineered lentiviral vectors in target cells. |
title_short |
Visualization of DC-SIGN-mediated entry pathway of engineered lentiviral vectors in target cells. |
title_full |
Visualization of DC-SIGN-mediated entry pathway of engineered lentiviral vectors in target cells. |
title_fullStr |
Visualization of DC-SIGN-mediated entry pathway of engineered lentiviral vectors in target cells. |
title_full_unstemmed |
Visualization of DC-SIGN-mediated entry pathway of engineered lentiviral vectors in target cells. |
title_sort |
visualization of dc-sign-mediated entry pathway of engineered lentiviral vectors in target cells. |
publisher |
Public Library of Science (PLoS) |
series |
PLoS ONE |
issn |
1932-6203 |
publishDate |
2013-01-01 |
description |
Dendritic cells (DCs) are potent antigen-presenting cells and therefore have enormous potential as vaccine targets. We have previously developed an engineered lentiviral vector (LV) that is pseudotyped with a mutated Sindbis virus glycoprotein (SVGmu), which is capable of targeting DCs through Dendritic Cell-specific ICAM3-grabbing Nonintegrin (DC-SIGN), a receptor that is predominantly expressed by DCs. In this study, we aimed to elucidate the internalization and trafficking mechanisms of this viral vector system through direct visualization of GFP-Vpr-tagged viral particles in target DCs, which was further corroborated by drug inhibition and dominant-negative mutants of cellular proteins that regulate the endocytic traffic. We demonstrated that our engineered LVs enter the cell via receptor-mediated clathrin- and dynamin-dependent endocytosis. Microtubule networks were also involved in a productive infection. Viral vector fusion was low-pH-dependent and occurred in the early endosomal stage of the intracellular transport. Autophagy was also examined for its effect on transduction efficiency, and we observed that enhanced autophage activity reduced vector infectivity, while suppressed autophagy boosted transduction efficiency. This study shed some light on the internalization and trafficking mechanisms of DC-directed LVs and offers some strategies to further improve the efficiency of LV-mediated gene therapy. |
url |
http://europepmc.org/articles/PMC3696072?pdf=render |
work_keys_str_mv |
AT yarongliu visualizationofdcsignmediatedentrypathwayofengineeredlentiviralvectorsintargetcells AT apriltai visualizationofdcsignmediatedentrypathwayofengineeredlentiviralvectorsintargetcells AT kyeiljoo visualizationofdcsignmediatedentrypathwayofengineeredlentiviralvectorsintargetcells AT pinwang visualizationofdcsignmediatedentrypathwayofengineeredlentiviralvectorsintargetcells |
_version_ |
1725342376764899328 |