The culture of primary duck endothelial cells for the study of avian influenza
Abstract Background Endothelial cells play a major role in highly pathogenic avian influenza (HPAI) virus pathogenesis in gallinaceous poultry species (e.g. chicken, turkey and quail). Upon infection of gallinaceous poultry with HPAI viruses, endothelial cells throughout the body become rapidly infe...
Main Authors: | , , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
BMC
2018-10-01
|
Series: | BMC Microbiology |
Subjects: | |
Online Access: | http://link.springer.com/article/10.1186/s12866-018-1307-4 |
id |
doaj-e4600d4d0383412790c398d75f49dc57 |
---|---|
record_format |
Article |
spelling |
doaj-e4600d4d0383412790c398d75f49dc572020-11-25T01:28:59ZengBMCBMC Microbiology1471-21802018-10-011811910.1186/s12866-018-1307-4The culture of primary duck endothelial cells for the study of avian influenzaRaissa L. Davis0Geunho Choi1Thijs Kuiken2Pascale Quéré3Sascha Trapp4Kirsty R. Short5Mathilde Richard6Department of Viroscience, Erasmus Medical CentreDepartment of Viroscience, Erasmus Medical CentreDepartment of Viroscience, Erasmus Medical CentreINRA ISP, Université de ToursINRA ISP, Université de ToursDepartment of Viroscience, Erasmus Medical CentreDepartment of Viroscience, Erasmus Medical CentreAbstract Background Endothelial cells play a major role in highly pathogenic avian influenza (HPAI) virus pathogenesis in gallinaceous poultry species (e.g. chicken, turkey and quail). Upon infection of gallinaceous poultry with HPAI viruses, endothelial cells throughout the body become rapidly infected, leading to systemic dissemination of the virus, disseminated intravascular coagulation, oedema and haemorrhaging. In contrast, the pathogenesis of HPAI viruses in most wild bird species (e.g. duck, goose and gull species) is not associated with endothelial tropism. Indeed, viral antigen is not found in the endothelial cells of most wild bird species following infection with HPAI viruses. This differential endothelial cell tropism in avian species is poorly understood, mainly due to the absence of appropriate cell culture systems. Results Here, we describe the isolation and purification of primary duck endothelial cells from the aorta or bone marrow of Pekin duck embryos. Cells were differentiated in the presence of vascular endothelial growth factor and, if needed, enriched via fluorescent-activated cell sorting based on the uptake of acetylated low-density lipoprotein. The expression of von Willebrand factor, a key marker of endothelial cells, was confirmed by polymerase chain reaction. Monocultures of duck endothelial cells, either derived from the aorta or the bone marrow, were susceptible to infection with an H5N1 HPAI virus but to a much lesser extent than chicken endothelial cells. Conclusions The methods described herein to isolate and purify duck endothelial cells from the aorta or bone marrow could also be applied to obtain microvascular endothelial cells from other tissues and organs, such as the lung or the intestine, and represent a valuable tool to study the pathogenesis of avian viruses.http://link.springer.com/article/10.1186/s12866-018-1307-4DuckEndothelial cellsHighly pathogenic avian influenza virus |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Raissa L. Davis Geunho Choi Thijs Kuiken Pascale Quéré Sascha Trapp Kirsty R. Short Mathilde Richard |
spellingShingle |
Raissa L. Davis Geunho Choi Thijs Kuiken Pascale Quéré Sascha Trapp Kirsty R. Short Mathilde Richard The culture of primary duck endothelial cells for the study of avian influenza BMC Microbiology Duck Endothelial cells Highly pathogenic avian influenza virus |
author_facet |
Raissa L. Davis Geunho Choi Thijs Kuiken Pascale Quéré Sascha Trapp Kirsty R. Short Mathilde Richard |
author_sort |
Raissa L. Davis |
title |
The culture of primary duck endothelial cells for the study of avian influenza |
title_short |
The culture of primary duck endothelial cells for the study of avian influenza |
title_full |
The culture of primary duck endothelial cells for the study of avian influenza |
title_fullStr |
The culture of primary duck endothelial cells for the study of avian influenza |
title_full_unstemmed |
The culture of primary duck endothelial cells for the study of avian influenza |
title_sort |
culture of primary duck endothelial cells for the study of avian influenza |
publisher |
BMC |
series |
BMC Microbiology |
issn |
1471-2180 |
publishDate |
2018-10-01 |
description |
Abstract Background Endothelial cells play a major role in highly pathogenic avian influenza (HPAI) virus pathogenesis in gallinaceous poultry species (e.g. chicken, turkey and quail). Upon infection of gallinaceous poultry with HPAI viruses, endothelial cells throughout the body become rapidly infected, leading to systemic dissemination of the virus, disseminated intravascular coagulation, oedema and haemorrhaging. In contrast, the pathogenesis of HPAI viruses in most wild bird species (e.g. duck, goose and gull species) is not associated with endothelial tropism. Indeed, viral antigen is not found in the endothelial cells of most wild bird species following infection with HPAI viruses. This differential endothelial cell tropism in avian species is poorly understood, mainly due to the absence of appropriate cell culture systems. Results Here, we describe the isolation and purification of primary duck endothelial cells from the aorta or bone marrow of Pekin duck embryos. Cells were differentiated in the presence of vascular endothelial growth factor and, if needed, enriched via fluorescent-activated cell sorting based on the uptake of acetylated low-density lipoprotein. The expression of von Willebrand factor, a key marker of endothelial cells, was confirmed by polymerase chain reaction. Monocultures of duck endothelial cells, either derived from the aorta or the bone marrow, were susceptible to infection with an H5N1 HPAI virus but to a much lesser extent than chicken endothelial cells. Conclusions The methods described herein to isolate and purify duck endothelial cells from the aorta or bone marrow could also be applied to obtain microvascular endothelial cells from other tissues and organs, such as the lung or the intestine, and represent a valuable tool to study the pathogenesis of avian viruses. |
topic |
Duck Endothelial cells Highly pathogenic avian influenza virus |
url |
http://link.springer.com/article/10.1186/s12866-018-1307-4 |
work_keys_str_mv |
AT raissaldavis thecultureofprimaryduckendothelialcellsforthestudyofavianinfluenza AT geunhochoi thecultureofprimaryduckendothelialcellsforthestudyofavianinfluenza AT thijskuiken thecultureofprimaryduckendothelialcellsforthestudyofavianinfluenza AT pascalequere thecultureofprimaryduckendothelialcellsforthestudyofavianinfluenza AT saschatrapp thecultureofprimaryduckendothelialcellsforthestudyofavianinfluenza AT kirstyrshort thecultureofprimaryduckendothelialcellsforthestudyofavianinfluenza AT mathilderichard thecultureofprimaryduckendothelialcellsforthestudyofavianinfluenza AT raissaldavis cultureofprimaryduckendothelialcellsforthestudyofavianinfluenza AT geunhochoi cultureofprimaryduckendothelialcellsforthestudyofavianinfluenza AT thijskuiken cultureofprimaryduckendothelialcellsforthestudyofavianinfluenza AT pascalequere cultureofprimaryduckendothelialcellsforthestudyofavianinfluenza AT saschatrapp cultureofprimaryduckendothelialcellsforthestudyofavianinfluenza AT kirstyrshort cultureofprimaryduckendothelialcellsforthestudyofavianinfluenza AT mathilderichard cultureofprimaryduckendothelialcellsforthestudyofavianinfluenza |
_version_ |
1725099208469381120 |