HIV-1-derived single-stranded RNA acts as activator of human neutrophils

Neutrophils are key effector cells of the innate immune system and are involved in the host defense against invading pathogens such as viruses. Recently, it was reported that HIV-1-neutrophil interaction triggers neutrophil activation and promotes expression of Toll-like receptors (TLR). Here, we as...

Full description

Autores:
Giraldo, Diana Marcela
Hernández López, Juan Carlos
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:20.500.12494/1192
Acceso en línea:
https://hdl.handle.net/20.500.12494/1192
Palabra clave:
HIV-1
Neutrophils
Rights
embargoedAccess
License
Licencia CC
id COOPER2_c8d716fbd16a49f154d21de8c6ed50b3
oai_identifier_str oai:repository.ucc.edu.co:20.500.12494/1192
network_acronym_str COOPER2
network_name_str Repositorio UCC
repository_id_str
dc.title.spa.fl_str_mv HIV-1-derived single-stranded RNA acts as activator of human neutrophils
title HIV-1-derived single-stranded RNA acts as activator of human neutrophils
spellingShingle HIV-1-derived single-stranded RNA acts as activator of human neutrophils
HIV-1
Neutrophils
title_short HIV-1-derived single-stranded RNA acts as activator of human neutrophils
title_full HIV-1-derived single-stranded RNA acts as activator of human neutrophils
title_fullStr HIV-1-derived single-stranded RNA acts as activator of human neutrophils
title_full_unstemmed HIV-1-derived single-stranded RNA acts as activator of human neutrophils
title_sort HIV-1-derived single-stranded RNA acts as activator of human neutrophils
dc.creator.fl_str_mv Giraldo, Diana Marcela
Hernández López, Juan Carlos
Urucqui Inchima, Silvio
dc.contributor.author.none.fl_str_mv Giraldo, Diana Marcela
Hernández López, Juan Carlos
Urucqui Inchima, Silvio
dc.subject.spa.fl_str_mv HIV-1
Neutrophils
topic HIV-1
Neutrophils
description Neutrophils are key effector cells of the innate immune system and are involved in the host defense against invading pathogens such as viruses. Recently, it was reported that HIV-1-neutrophil interaction triggers neutrophil activation and promotes expression of Toll-like receptors (TLR). Here, we assessed the role of single-stranded RNA40 (ssRNA40) derived from HIV-1 in neutrophil activation. We observed functional activation of neutrophils in response to HIV-1-derived ssRNA40 based on the expression of TLR7/8, RIG-I and MDA5, induction of cytokines (IL-6 and TNF-α) and the production of reactive oxygen species (ROS). Additionally, ssRNA40 promoted the expression of CD62L and TNF-α and the production of ROS in the presence of the TLR2 agonist Pam2CSK4. ssRNA40 together with R848 (an RIG-I/MDA5 agonist) increased CD11b expression but decreased CD62L expression. Furthermore, decreased IL-6 expression was observed in the presence of the TLR4 agonist LPS. Finally, we found that ssRNA40 promotes RIG-I and MDA5 expression in the presence of the TLR2, TLR4 and TLR7/8 agonists. This study demonstrates a functional response of TLRs in neutrophils challenged with ssRNA40, suggesting that TLRs could be involved in the innate immune response observed during HIV infection, which might be mediated by its genome.
publishDate 2016
dc.date.issued.none.fl_str_mv 2016-10
dc.date.accessioned.none.fl_str_mv 2017-08-19T00:07:01Z
dc.date.available.none.fl_str_mv 2017-10
2017-08-19T00:07:01Z
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/20.500.12494/1192
dc.identifier.bibliographicCitation.spa.fl_str_mv Giraldo D. M., Hernández, J. C.& Urcuqui Inchima, S. (2016). HIV-1-derived single-stranded RNA acts as activator of human neutrophils. Immunologic Research, 64 (5). 1185-1194. doi: 10.1007/s12026-016-8876-9
url https://hdl.handle.net/20.500.12494/1192
identifier_str_mv Giraldo D. M., Hernández, J. C.& Urcuqui Inchima, S. (2016). HIV-1-derived single-stranded RNA acts as activator of human neutrophils. Immunologic Research, 64 (5). 1185-1194. doi: 10.1007/s12026-016-8876-9
dc.relation.isversionof.spa.fl_str_mv https://link.springer.com/article/10.1007%2Fs12026-016-8876-9
dc.rights.cc.none.fl_str_mv Licencia CC
dc.rights.accessrights.none.fl_str_mv info:eu-repo/semantics/embargoedAccess
dc.rights.coar.none.fl_str_mv http://purl.org/coar/access_right/c_f1cf
rights_invalid_str_mv Licencia CC
http://purl.org/coar/access_right/c_f1cf
eu_rights_str_mv embargoedAccess
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/00b1394c-715c-4153-ad89-c6f0f612fefe/download
https://repository.ucc.edu.co/bitstreams/52349aa2-ff83-4713-932f-6a5c8710c2eb/download
https://repository.ucc.edu.co/bitstreams/591b76c0-111b-48ec-b6e5-ee4a427e45c7/download
https://repository.ucc.edu.co/bitstreams/8318365d-aac5-44b6-a18f-6581146e574a/download
bitstream.checksum.fl_str_mv c9581f4e9abb8b7d319368a8dacba226
cb8f4d9b72d863ad6737f22af442cc82
59a8e8912d130855d800ef628764056e
b2a8ae1b66463f41e4faacd3bf1b1713
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_ 1814247311884156928
spelling Giraldo, Diana MarcelaHernández López, Juan CarlosUrucqui Inchima, Silvio2017-08-19T00:07:01Z2017-102017-08-19T00:07:01Z2016-10https://hdl.handle.net/20.500.12494/1192Giraldo D. M., Hernández, J. C.& Urcuqui Inchima, S. (2016). HIV-1-derived single-stranded RNA acts as activator of human neutrophils. Immunologic Research, 64 (5). 1185-1194. doi: 10.1007/s12026-016-8876-9Neutrophils are key effector cells of the innate immune system and are involved in the host defense against invading pathogens such as viruses. Recently, it was reported that HIV-1-neutrophil interaction triggers neutrophil activation and promotes expression of Toll-like receptors (TLR). Here, we assessed the role of single-stranded RNA40 (ssRNA40) derived from HIV-1 in neutrophil activation. We observed functional activation of neutrophils in response to HIV-1-derived ssRNA40 based on the expression of TLR7/8, RIG-I and MDA5, induction of cytokines (IL-6 and TNF-α) and the production of reactive oxygen species (ROS). Additionally, ssRNA40 promoted the expression of CD62L and TNF-α and the production of ROS in the presence of the TLR2 agonist Pam2CSK4. ssRNA40 together with R848 (an RIG-I/MDA5 agonist) increased CD11b expression but decreased CD62L expression. Furthermore, decreased IL-6 expression was observed in the presence of the TLR4 agonist LPS. Finally, we found that ssRNA40 promotes RIG-I and MDA5 expression in the presence of the TLR2, TLR4 and TLR7/8 agonists. This study demonstrates a functional response of TLRs in neutrophils challenged with ssRNA40, suggesting that TLRs could be involved in the innate immune response observed during HIV infection, which might be mediated by its genome.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%2Fs12026-016-8876-9HIV-1NeutrophilsHIV-1-derived single-stranded RNA acts as activator of human neutrophilsArtículohttp://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/embargoedAccesshttp://purl.org/coar/access_right/c_f1cfPublicationORIGINALpre-print ssRNA.pdfpre-print ssRNA.pdfapplication/pdf578218https://repository.ucc.edu.co/bitstreams/00b1394c-715c-4153-ad89-c6f0f612fefe/downloadc9581f4e9abb8b7d319368a8dacba226MD51LICENSElicense.txtlicense.txttext/plain; charset=utf-84553https://repository.ucc.edu.co/bitstreams/52349aa2-ff83-4713-932f-6a5c8710c2eb/downloadcb8f4d9b72d863ad6737f22af442cc82MD52TEXTpre-print ssRNA.pdf.txtpre-print ssRNA.pdf.txtExtracted texttext/plain40997https://repository.ucc.edu.co/bitstreams/591b76c0-111b-48ec-b6e5-ee4a427e45c7/download59a8e8912d130855d800ef628764056eMD53THUMBNAILpre-print ssRNA.pdf.jpgpre-print ssRNA.pdf.jpgIM Thumbnailimage/jpeg4360https://repository.ucc.edu.co/bitstreams/8318365d-aac5-44b6-a18f-6581146e574a/downloadb2a8ae1b66463f41e4faacd3bf1b1713MD5420.500.12494/1192oai:repository.ucc.edu.co:20.500.12494/11922024-08-10 22:40:34.72open.accesshttps://repository.ucc.edu.coRepositorio Institucional Universidad Cooperativa de Colombiabdigital@metabiblioteca.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