Recombinant hepatitis C virus-envelope protein 2 interactions with low-density lipoprotein/CD81 receptors
Autor(a) principal: | |
---|---|
Data de Publicação: | 2015 |
Outros Autores: | , , , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762015000400534&lng=en&nrm=iso&tlng=en http://hdl.handle.net/11449/129365 |
Resumo: | Hepatitis C virus (HCV) envelope protein 2 (E2) is involved in viral binding to host cells. The aim of this work was to produce recombinant E2B and E2Y HCV proteins in Escherichia coli and Pichia pastoris, respectively, and to study their interactions with low-density lipoprotein receptor (LDLr) and CD81 in human umbilical vein endothelial cells (HUVEC) and the ECV304 bladder carcinoma cell line. To investigate the effects of human LDL and differences in protein structure (glycosylated or not) on binding efficiency, the recombinant proteins were either associated or not associated with lipoproteins before being assayed. The immunoreactivity of the recombinant proteins was analysed using pooled serum samples that were either positive or negative for hepatitis C. The cells were immunophenotyped by LDLr and CD81 using flow cytometry. Binding and binding inhibition assays were performed in the presence of LDL, foetal bovine serum (FCS) and specific antibodies. The results revealed that binding was reduced in the absence of FCS, but that the addition of human LDL rescued and increased binding capacity. In HUVEC cells, the use of antibodies to block LDLr led to a significant reduction in the binding of E2B and E2Y. CD81 antibodies did not affect E2B and E2Y binding. In ECV304 cells, blocking LDLr and CD81 produced similar effects, but they were not as marked as those that were observed in HUVEC cells. In conclusion, recombinant HCV E2 is dependent on LDL for its ability to bind to LDLr in HUVEC and ECV304 cells. These findings are relevant because E2 acts to anchor HCV to host cells; therefore, high blood levels of LDL could enhance viral infectivity in chronic hepatitis C patients. |
id |
UNSP_e21b4be82539122f93fc7832b6b68346 |
---|---|
oai_identifier_str |
oai:repositorio.unesp.br:11449/129365 |
network_acronym_str |
UNSP |
network_name_str |
Repositório Institucional da UNESP |
repository_id_str |
2946 |
spelling |
Recombinant hepatitis C virus-envelope protein 2 interactions with low-density lipoprotein/CD81 receptorsHCVE2LDLrCD81Hepatitis C virus (HCV) envelope protein 2 (E2) is involved in viral binding to host cells. The aim of this work was to produce recombinant E2B and E2Y HCV proteins in Escherichia coli and Pichia pastoris, respectively, and to study their interactions with low-density lipoprotein receptor (LDLr) and CD81 in human umbilical vein endothelial cells (HUVEC) and the ECV304 bladder carcinoma cell line. To investigate the effects of human LDL and differences in protein structure (glycosylated or not) on binding efficiency, the recombinant proteins were either associated or not associated with lipoproteins before being assayed. The immunoreactivity of the recombinant proteins was analysed using pooled serum samples that were either positive or negative for hepatitis C. The cells were immunophenotyped by LDLr and CD81 using flow cytometry. Binding and binding inhibition assays were performed in the presence of LDL, foetal bovine serum (FCS) and specific antibodies. The results revealed that binding was reduced in the absence of FCS, but that the addition of human LDL rescued and increased binding capacity. In HUVEC cells, the use of antibodies to block LDLr led to a significant reduction in the binding of E2B and E2Y. CD81 antibodies did not affect E2B and E2Y binding. In ECV304 cells, blocking LDLr and CD81 produced similar effects, but they were not as marked as those that were observed in HUVEC cells. In conclusion, recombinant HCV E2 is dependent on LDL for its ability to bind to LDLr in HUVEC and ECV304 cells. These findings are relevant because E2 acts to anchor HCV to host cells; therefore, high blood levels of LDL could enhance viral infectivity in chronic hepatitis C patients.Univ Estadual Paulista, Fac Ciencias Farmaceut Araraquara, Dept Anal Clin, Bauru, SP, BrazilUniv Estadual Paulista, Fac Ciencias, Dept Quim, Bauru, SP, BrazilUniv Nova Lisboa, Inst Higiene &Med Trop, Unidade Parasitol Med &Microbiol, Dept Parasitol Med, P-1200 Lisbon, PortugalUniv Fed Sao Carlos, Dept Morfol &Patol, BR-13560 Sao Carlos, SP, BrazilUniv Fed Sao Carlos, Dept Genet &Evolucao, BR-13560 Sao Carlos, SP, BrazilUniv Sao Paulo, Inst Quim Sao Carlos, Dept Quim &Fis Mol, Sao Carlos, SP, BrazilUniv Estadual Paulista, Fac Ciencias Farmaceut Araraquara, Dept Anal Clin, Bauru, SP, BrazilUniv Estadual Paulista, Fac Ciencias, Dept Quim, Bauru, SP, BrazilFundaco Oswaldo CruzUniversidade Estadual Paulista (Unesp)Univ Nova LisboaUniversidade Federal de São Carlos (UFSCar)Universidade de São Paulo (USP)Urbaczek, Ana Carolina [UNESP]Ximenes, Valdecir Farias [UNESP]Afonso, AnaGeneroso, Wesley CardosoNogueira, Camila Tita [UNESP]Tansini, Aline [UNESP]Dias Cappelini, Luciana TeresaMalago Junior, WilsonSilva, Flavio Henrique daFonseca, Luiz Marcos da [UNESP]Costa, Paulo Inacio da [UNESP]2015-10-21T20:56:09Z2015-10-21T20:56:09Z2015-06-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article534-542application/pdfhttp://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762015000400534&lng=en&nrm=iso&tlng=enMemorias do Instituto Oswaldo Cruz, v. 110, n. 4, p. 534-542, 2015.0074-0276http://hdl.handle.net/11449/12936510.1590/0074-02760140441S0074-02762015000400534WOS:000356610000012S0074-02762015000400534.pdf406641399790857267202237159173810000-0002-3350-8308Web of Sciencereponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengMemorias do Instituto Oswaldo Cruz2.8331,172info:eu-repo/semantics/openAccess2024-06-21T15:19:08Zoai:repositorio.unesp.br:11449/129365Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-06-21T15:19:08Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Recombinant hepatitis C virus-envelope protein 2 interactions with low-density lipoprotein/CD81 receptors |
title |
Recombinant hepatitis C virus-envelope protein 2 interactions with low-density lipoprotein/CD81 receptors |
spellingShingle |
Recombinant hepatitis C virus-envelope protein 2 interactions with low-density lipoprotein/CD81 receptors Urbaczek, Ana Carolina [UNESP] HCV E2 LDLr CD81 |
title_short |
Recombinant hepatitis C virus-envelope protein 2 interactions with low-density lipoprotein/CD81 receptors |
title_full |
Recombinant hepatitis C virus-envelope protein 2 interactions with low-density lipoprotein/CD81 receptors |
title_fullStr |
Recombinant hepatitis C virus-envelope protein 2 interactions with low-density lipoprotein/CD81 receptors |
title_full_unstemmed |
Recombinant hepatitis C virus-envelope protein 2 interactions with low-density lipoprotein/CD81 receptors |
title_sort |
Recombinant hepatitis C virus-envelope protein 2 interactions with low-density lipoprotein/CD81 receptors |
author |
Urbaczek, Ana Carolina [UNESP] |
author_facet |
Urbaczek, Ana Carolina [UNESP] Ximenes, Valdecir Farias [UNESP] Afonso, Ana Generoso, Wesley Cardoso Nogueira, Camila Tita [UNESP] Tansini, Aline [UNESP] Dias Cappelini, Luciana Teresa Malago Junior, Wilson Silva, Flavio Henrique da Fonseca, Luiz Marcos da [UNESP] Costa, Paulo Inacio da [UNESP] |
author_role |
author |
author2 |
Ximenes, Valdecir Farias [UNESP] Afonso, Ana Generoso, Wesley Cardoso Nogueira, Camila Tita [UNESP] Tansini, Aline [UNESP] Dias Cappelini, Luciana Teresa Malago Junior, Wilson Silva, Flavio Henrique da Fonseca, Luiz Marcos da [UNESP] Costa, Paulo Inacio da [UNESP] |
author2_role |
author author author author author author author author author author |
dc.contributor.none.fl_str_mv |
Universidade Estadual Paulista (Unesp) Univ Nova Lisboa Universidade Federal de São Carlos (UFSCar) Universidade de São Paulo (USP) |
dc.contributor.author.fl_str_mv |
Urbaczek, Ana Carolina [UNESP] Ximenes, Valdecir Farias [UNESP] Afonso, Ana Generoso, Wesley Cardoso Nogueira, Camila Tita [UNESP] Tansini, Aline [UNESP] Dias Cappelini, Luciana Teresa Malago Junior, Wilson Silva, Flavio Henrique da Fonseca, Luiz Marcos da [UNESP] Costa, Paulo Inacio da [UNESP] |
dc.subject.por.fl_str_mv |
HCV E2 LDLr CD81 |
topic |
HCV E2 LDLr CD81 |
description |
Hepatitis C virus (HCV) envelope protein 2 (E2) is involved in viral binding to host cells. The aim of this work was to produce recombinant E2B and E2Y HCV proteins in Escherichia coli and Pichia pastoris, respectively, and to study their interactions with low-density lipoprotein receptor (LDLr) and CD81 in human umbilical vein endothelial cells (HUVEC) and the ECV304 bladder carcinoma cell line. To investigate the effects of human LDL and differences in protein structure (glycosylated or not) on binding efficiency, the recombinant proteins were either associated or not associated with lipoproteins before being assayed. The immunoreactivity of the recombinant proteins was analysed using pooled serum samples that were either positive or negative for hepatitis C. The cells were immunophenotyped by LDLr and CD81 using flow cytometry. Binding and binding inhibition assays were performed in the presence of LDL, foetal bovine serum (FCS) and specific antibodies. The results revealed that binding was reduced in the absence of FCS, but that the addition of human LDL rescued and increased binding capacity. In HUVEC cells, the use of antibodies to block LDLr led to a significant reduction in the binding of E2B and E2Y. CD81 antibodies did not affect E2B and E2Y binding. In ECV304 cells, blocking LDLr and CD81 produced similar effects, but they were not as marked as those that were observed in HUVEC cells. In conclusion, recombinant HCV E2 is dependent on LDL for its ability to bind to LDLr in HUVEC and ECV304 cells. These findings are relevant because E2 acts to anchor HCV to host cells; therefore, high blood levels of LDL could enhance viral infectivity in chronic hepatitis C patients. |
publishDate |
2015 |
dc.date.none.fl_str_mv |
2015-10-21T20:56:09Z 2015-10-21T20:56:09Z 2015-06-01 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762015000400534&lng=en&nrm=iso&tlng=en Memorias do Instituto Oswaldo Cruz, v. 110, n. 4, p. 534-542, 2015. 0074-0276 http://hdl.handle.net/11449/129365 10.1590/0074-02760140441 S0074-02762015000400534 WOS:000356610000012 S0074-02762015000400534.pdf 4066413997908572 6720223715917381 0000-0002-3350-8308 |
url |
http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762015000400534&lng=en&nrm=iso&tlng=en http://hdl.handle.net/11449/129365 |
identifier_str_mv |
Memorias do Instituto Oswaldo Cruz, v. 110, n. 4, p. 534-542, 2015. 0074-0276 10.1590/0074-02760140441 S0074-02762015000400534 WOS:000356610000012 S0074-02762015000400534.pdf 4066413997908572 6720223715917381 0000-0002-3350-8308 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Memorias do Instituto Oswaldo Cruz 2.833 1,172 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
534-542 application/pdf |
dc.publisher.none.fl_str_mv |
Fundaco Oswaldo Cruz |
publisher.none.fl_str_mv |
Fundaco Oswaldo Cruz |
dc.source.none.fl_str_mv |
Web of Science reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
collection |
Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
|
_version_ |
1803649907805913088 |