HIV-1-exposed seronegative individuals show alteration in TLR expression and pro-inflammatory cytokine production ex vivo: An innate immune quiescence status?

Pattern recognition receptors (PRRs) are involved in direct recognition of viruses, promoting cellular activation and the production of pro-inflammatory cytokines. However, despite the reduced systemic immune activation described in HIV-1-exposed seronegatives (HESNs), few studies have focused on de...

Full description

Autores:
Hernández López, Juan Carlos
St. Laurent III, Georges
Urucqui Inchima, Silvio
Tipo de recurso:
Article of journal
Fecha de publicación:
2016
Institución:
Universidad Cooperativa de Colombia
Repositorio:
Repositorio UCC
Idioma:
OAI Identifier:
oai:repository.ucc.edu.co:20.500.12494/1185
Acceso en línea:
https://hdl.handle.net/20.500.12494/1185
Palabra clave:
HIV-1
Inflammation
Rights
openAccess
License
Licencia CC
id COOPER2_a6fe1903b88f2ea2309a813ca7eea3fd
oai_identifier_str oai:repository.ucc.edu.co:20.500.12494/1185
network_acronym_str COOPER2
network_name_str Repositorio UCC
repository_id_str
dc.title.spa.fl_str_mv HIV-1-exposed seronegative individuals show alteration in TLR expression and pro-inflammatory cytokine production ex vivo: An innate immune quiescence status?
title HIV-1-exposed seronegative individuals show alteration in TLR expression and pro-inflammatory cytokine production ex vivo: An innate immune quiescence status?
spellingShingle HIV-1-exposed seronegative individuals show alteration in TLR expression and pro-inflammatory cytokine production ex vivo: An innate immune quiescence status?
HIV-1
Inflammation
title_short HIV-1-exposed seronegative individuals show alteration in TLR expression and pro-inflammatory cytokine production ex vivo: An innate immune quiescence status?
title_full HIV-1-exposed seronegative individuals show alteration in TLR expression and pro-inflammatory cytokine production ex vivo: An innate immune quiescence status?
title_fullStr HIV-1-exposed seronegative individuals show alteration in TLR expression and pro-inflammatory cytokine production ex vivo: An innate immune quiescence status?
title_full_unstemmed HIV-1-exposed seronegative individuals show alteration in TLR expression and pro-inflammatory cytokine production ex vivo: An innate immune quiescence status?
title_sort HIV-1-exposed seronegative individuals show alteration in TLR expression and pro-inflammatory cytokine production ex vivo: An innate immune quiescence status?
dc.creator.fl_str_mv Hernández López, Juan Carlos
St. Laurent III, Georges
Urucqui Inchima, Silvio
dc.contributor.author.none.fl_str_mv Hernández López, Juan Carlos
St. Laurent III, Georges
Urucqui Inchima, Silvio
dc.subject.spa.fl_str_mv HIV-1
Inflammation
topic HIV-1
Inflammation
description Pattern recognition receptors (PRRs) are involved in direct recognition of viruses, promoting cellular activation and the production of pro-inflammatory cytokines. However, despite the reduced systemic immune activation described in HIV-1-exposed seronegatives (HESNs), few studies have focused on determining the relationship between PRR expression and cytokine production. We have aimed here to evaluate the expression level of PRRs and cytokines in HESNs, HIV-1 patients and healthy donors. Basal PRR expression levels in PBMCs, dendritic cells (DCs) and monocytes, and plasma cytokine levels as well as the PRR ligand-induced cytokine productions were determined by flow cytometry, qPCR and ELISA. Higher TLR2/4 expression in DCs and monocytes from HESNs was observed. Nevertheless, TLR4/8, NOD2 and RIG-I mRNA levels were lower in PBMCs from HESNs than HIV-1-infected patients. Comparable IL-1b, IL-18 and TNF-a mRNA levels were observed among the groups examined; however, at the protein level, production of IL-1b, IL-6 and IL-10 was significantly lower in plasma from HESNs than from HIV-1-infected patients. Our results suggest that exposure to HIV-1 without infection could be associated with reduced basal proinflammatory responses. Further studies are required to define the cell subsets responsible for these differences and the role of PRRs on protection against HIV-1 infection.
publishDate 2016
dc.date.issued.none.fl_str_mv 2016-02
dc.date.accessioned.none.fl_str_mv 2017-08-18T23:22:11Z
dc.date.available.none.fl_str_mv 2017-08-18T23:22:11Z
dc.type.none.fl_str_mv Artículo
dc.type.coar.fl_str_mv http://purl.org/coar/resource_type/c_2df8fbb1
dc.type.coar.none.fl_str_mv http://purl.org/coar/resource_type/c_6501
dc.type.coarversion.none.fl_str_mv http://purl.org/coar/version/c_970fb48d4fbd8a85
dc.type.driver.none.fl_str_mv info:eu-repo/semantics/article
dc.type.version.none.fl_str_mv info:eu-repo/semantics/publishedVersion
format http://purl.org/coar/resource_type/c_6501
status_str publishedVersion
dc.identifier.uri.none.fl_str_mv https://hdl.handle.net/20.500.12494/1185
dc.identifier.bibliographicCitation.spa.fl_str_mv Hernández, J. C., St. Laurent 3rd, Urcuqui Inchima, S. (2016). HIV-1-exposed seronegative individuals show alteration in TLR expression and pro-inflammatory cytokine production ex vivo: An innate immune quiescence status? Immunologic Research, 64(1), 280-290. doi: 10.1007/s12026-015-8748-8.
url https://hdl.handle.net/20.500.12494/1185
identifier_str_mv Hernández, J. C., St. Laurent 3rd, Urcuqui Inchima, S. (2016). HIV-1-exposed seronegative individuals show alteration in TLR expression and pro-inflammatory cytokine production ex vivo: An innate immune quiescence status? Immunologic Research, 64(1), 280-290. doi: 10.1007/s12026-015-8748-8.
dc.relation.isversionof.spa.fl_str_mv https://link.springer.com/article/10.1007%2Fs12026-015-8748-8
dc.rights.cc.none.fl_str_mv Licencia CC
dc.rights.accessrights.none.fl_str_mv info:eu-repo/semantics/openAccess
dc.rights.coar.none.fl_str_mv http://purl.org/coar/access_right/c_abf2
rights_invalid_str_mv Licencia CC
http://purl.org/coar/access_right/c_abf2
eu_rights_str_mv openAccess
dc.publisher.spa.fl_str_mv Universidad Cooperativa de Colombia, Facultad de Ciencias de la Salud, Programa de Medicina, Medellín y Envigado, Colombia, 00000
dc.publisher.program.spa.fl_str_mv Medicina
dc.publisher.place.spa.fl_str_mv Medellín
institution Universidad Cooperativa de Colombia
bitstream.url.fl_str_mv https://repository.ucc.edu.co/bitstreams/6601abf1-16aa-40ed-ac50-459a09c3f908/download
https://repository.ucc.edu.co/bitstreams/88aad2a3-cdc7-481c-ba5a-a5b8d3d1a283/download
https://repository.ucc.edu.co/bitstreams/4c53f489-91d2-4c12-8f09-d7d76c7b992c/download
https://repository.ucc.edu.co/bitstreams/cfb11d8e-db3f-42ce-9de6-7a5f734d3bf7/download
bitstream.checksum.fl_str_mv cb8f4d9b72d863ad6737f22af442cc82
2988b493af59b56566ff4101c723797b
0cdf8b8bdb596567273e55eb7dba4147
caa9b23cc3f4de64dcd7d66fdab7146a
bitstream.checksumAlgorithm.fl_str_mv MD5
MD5
MD5
MD5
repository.name.fl_str_mv Repositorio Institucional Universidad Cooperativa de Colombia
repository.mail.fl_str_mv bdigital@metabiblioteca.com
_version_ 1808789236253458432
spelling Hernández López, Juan CarlosSt. Laurent III, GeorgesUrucqui Inchima, Silvio2017-08-18T23:22:11Z2017-08-18T23:22:11Z2016-02https://hdl.handle.net/20.500.12494/1185Hernández, J. C., St. Laurent 3rd, Urcuqui Inchima, S. (2016). HIV-1-exposed seronegative individuals show alteration in TLR expression and pro-inflammatory cytokine production ex vivo: An innate immune quiescence status? Immunologic Research, 64(1), 280-290. doi: 10.1007/s12026-015-8748-8.Pattern recognition receptors (PRRs) are involved in direct recognition of viruses, promoting cellular activation and the production of pro-inflammatory cytokines. However, despite the reduced systemic immune activation described in HIV-1-exposed seronegatives (HESNs), few studies have focused on determining the relationship between PRR expression and cytokine production. We have aimed here to evaluate the expression level of PRRs and cytokines in HESNs, HIV-1 patients and healthy donors. Basal PRR expression levels in PBMCs, dendritic cells (DCs) and monocytes, and plasma cytokine levels as well as the PRR ligand-induced cytokine productions were determined by flow cytometry, qPCR and ELISA. Higher TLR2/4 expression in DCs and monocytes from HESNs was observed. Nevertheless, TLR4/8, NOD2 and RIG-I mRNA levels were lower in PBMCs from HESNs than HIV-1-infected patients. Comparable IL-1b, IL-18 and TNF-a mRNA levels were observed among the groups examined; however, at the protein level, production of IL-1b, IL-6 and IL-10 was significantly lower in plasma from HESNs than from HIV-1-infected patients. Our results suggest that exposure to HIV-1 without infection could be associated with reduced basal proinflammatory responses. Further studies are required to define the cell subsets responsible for these differences and the role of PRRs on protection against HIV-1 infection.juanc.hernandezl@campusucc.edu.coUniversidad Cooperativa de Colombia, Facultad de Ciencias de la Salud, Programa de Medicina, Medellín y Envigado, Colombia, 00000MedicinaMedellínhttps://link.springer.com/article/10.1007%2Fs12026-015-8748-8HIV-1InflammationHIV-1-exposed seronegative individuals show alteration in TLR expression and pro-inflammatory cytokine production ex vivo: An innate immune quiescence status?Artículohttp://purl.org/coar/resource_type/c_6501http://purl.org/coar/resource_type/c_2df8fbb1http://purl.org/coar/version/c_970fb48d4fbd8a85info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionLicencia CCinfo:eu-repo/semantics/openAccesshttp://purl.org/coar/access_right/c_abf2PublicationLICENSElicense.txtlicense.txttext/plain; charset=utf-84553https://repository.ucc.edu.co/bitstreams/6601abf1-16aa-40ed-ac50-459a09c3f908/downloadcb8f4d9b72d863ad6737f22af442cc82MD52TEXTpost-print.pdf.txtpost-print.pdf.txtExtracted texttext/plain44442https://repository.ucc.edu.co/bitstreams/88aad2a3-cdc7-481c-ba5a-a5b8d3d1a283/download2988b493af59b56566ff4101c723797bMD53THUMBNAILpost-print.pdf.jpgpost-print.pdf.jpgIM Thumbnailimage/jpeg4072https://repository.ucc.edu.co/bitstreams/4c53f489-91d2-4c12-8f09-d7d76c7b992c/download0cdf8b8bdb596567273e55eb7dba4147MD54ORIGINALpost-print.pdfpost-print.pdfapplication/pdf549893https://repository.ucc.edu.co/bitstreams/cfb11d8e-db3f-42ce-9de6-7a5f734d3bf7/downloadcaa9b23cc3f4de64dcd7d66fdab7146aMD5120.500.12494/1185oai:repository.ucc.edu.co:20.500.12494/11852024-08-10 22:44:17.335open.accesshttps://repository.ucc.edu.coRepositorio Institucional Universidad Cooperativa de Colombiabdigital@metabiblioteca.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