Abstract
Amiodarone interferes with the endocytic pathway, inhibits proteolysis, and causes the formation of vacuoles, but uptake and intracellular distribution of the drug, origin of vacuoles, and functional consequences of amiodarone accumulation remain unclear. Our objective was to study amiodarone uptake, clarify the origin of vacuoles, and investigate the effect of amiodarone on the life cycle of the coronavirus responsible for the Severe Acute Respiratory Syndrome (SARS), which, to enter cells, relies on the proteolytic cleavage of a viral spike protein by the endosomal proteinase cathepsin L. Using alveolar macrophages, we studied uptake of 125I-amiodarone and 125I-B2, an analog lacking the lateral group diethylamino-β- ethoxy, and analyzed the effects of amiodarone on the distribution of endosomal markers and on the uptake of an acidotropic dye. Furthermore, using Vero cells, we tested the impact of amiodarone on the in vitro spreading of the SARS coronavirus. We found that (1) amiodarone associates with different cell membranes and accumulates in acidic organelles; (2) the diethylamino-β- ethoxy group is an important determinant of uptake; (3) vacuoles forming upon exposure to amiodarone are enlarged late endosomes; (4) amiodarone inhibits the spreading in vitro of SARS coronavirus; and (5) trypsin cleavage of the viral spike protein before infection, which permits virus entry through the plasma membrane, does not impair amiodarone antiviral activity. We conclude that amiodarone alters late compartments of the endocytic pathway and inhibits SARS coronavirus infection by acting after the transit of the virus through endosomes.
| Original language | English |
|---|---|
| Pages (from-to) | 142-149 |
| Number of pages | 8 |
| Journal | American Journal of Respiratory Cell and Molecular Biology |
| Volume | 39 |
| Issue number | 2 |
| DOIs | |
| Publication status | Published - Aug 1 2008 |
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Keywords
- Amiodarone
- Endocytic pathway
- SARS coronavirus
ASJC Scopus subject areas
- Cell Biology
- Pulmonary and Respiratory Medicine
- Molecular Biology
Cite this
Amiodarone alters late endosomes and inhibits SARS coronavirus infection at a post-endosomal level. / Stadler, Konrad; Ha, Huy Riem; Ciminale, Vincenzo; Spirli, Carlo; Saletti, Giulietta; Schiavon, Marco; Bruttomesso, Daniela; Bigler, Laurent; Follath, Ferenc; Pettenazzo, Andrea; Baritussio, Aldo.
In: American Journal of Respiratory Cell and Molecular Biology, Vol. 39, No. 2, 01.08.2008, p. 142-149.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Amiodarone alters late endosomes and inhibits SARS coronavirus infection at a post-endosomal level
AU - Stadler, Konrad
AU - Ha, Huy Riem
AU - Ciminale, Vincenzo
AU - Spirli, Carlo
AU - Saletti, Giulietta
AU - Schiavon, Marco
AU - Bruttomesso, Daniela
AU - Bigler, Laurent
AU - Follath, Ferenc
AU - Pettenazzo, Andrea
AU - Baritussio, Aldo
PY - 2008/8/1
Y1 - 2008/8/1
N2 - Amiodarone interferes with the endocytic pathway, inhibits proteolysis, and causes the formation of vacuoles, but uptake and intracellular distribution of the drug, origin of vacuoles, and functional consequences of amiodarone accumulation remain unclear. Our objective was to study amiodarone uptake, clarify the origin of vacuoles, and investigate the effect of amiodarone on the life cycle of the coronavirus responsible for the Severe Acute Respiratory Syndrome (SARS), which, to enter cells, relies on the proteolytic cleavage of a viral spike protein by the endosomal proteinase cathepsin L. Using alveolar macrophages, we studied uptake of 125I-amiodarone and 125I-B2, an analog lacking the lateral group diethylamino-β- ethoxy, and analyzed the effects of amiodarone on the distribution of endosomal markers and on the uptake of an acidotropic dye. Furthermore, using Vero cells, we tested the impact of amiodarone on the in vitro spreading of the SARS coronavirus. We found that (1) amiodarone associates with different cell membranes and accumulates in acidic organelles; (2) the diethylamino-β- ethoxy group is an important determinant of uptake; (3) vacuoles forming upon exposure to amiodarone are enlarged late endosomes; (4) amiodarone inhibits the spreading in vitro of SARS coronavirus; and (5) trypsin cleavage of the viral spike protein before infection, which permits virus entry through the plasma membrane, does not impair amiodarone antiviral activity. We conclude that amiodarone alters late compartments of the endocytic pathway and inhibits SARS coronavirus infection by acting after the transit of the virus through endosomes.
AB - Amiodarone interferes with the endocytic pathway, inhibits proteolysis, and causes the formation of vacuoles, but uptake and intracellular distribution of the drug, origin of vacuoles, and functional consequences of amiodarone accumulation remain unclear. Our objective was to study amiodarone uptake, clarify the origin of vacuoles, and investigate the effect of amiodarone on the life cycle of the coronavirus responsible for the Severe Acute Respiratory Syndrome (SARS), which, to enter cells, relies on the proteolytic cleavage of a viral spike protein by the endosomal proteinase cathepsin L. Using alveolar macrophages, we studied uptake of 125I-amiodarone and 125I-B2, an analog lacking the lateral group diethylamino-β- ethoxy, and analyzed the effects of amiodarone on the distribution of endosomal markers and on the uptake of an acidotropic dye. Furthermore, using Vero cells, we tested the impact of amiodarone on the in vitro spreading of the SARS coronavirus. We found that (1) amiodarone associates with different cell membranes and accumulates in acidic organelles; (2) the diethylamino-β- ethoxy group is an important determinant of uptake; (3) vacuoles forming upon exposure to amiodarone are enlarged late endosomes; (4) amiodarone inhibits the spreading in vitro of SARS coronavirus; and (5) trypsin cleavage of the viral spike protein before infection, which permits virus entry through the plasma membrane, does not impair amiodarone antiviral activity. We conclude that amiodarone alters late compartments of the endocytic pathway and inhibits SARS coronavirus infection by acting after the transit of the virus through endosomes.
KW - Amiodarone
KW - Endocytic pathway
KW - SARS coronavirus
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UR - http://www.scopus.com/inward/citedby.url?scp=48649102819&partnerID=8YFLogxK
U2 - 10.1165/rcmb.2007-0217OC
DO - 10.1165/rcmb.2007-0217OC
M3 - Article
C2 - 18314540
AN - SCOPUS:48649102819
VL - 39
SP - 142
EP - 149
JO - American Journal of Respiratory Cell and Molecular Biology
JF - American Journal of Respiratory Cell and Molecular Biology
SN - 1044-1549
IS - 2
ER -