Cargando…

A Tat/Rev Induced Limiting Dilution Assay to Measure Viral Reservoirs in Non-Human Primate Models of HIV Infection

The establishment of latent infection and poorly characterized viral reservoirs in tissues represent major obstacles to a definitive cure for HIV. Non-human primate (NHP) models of HIV infection are critical to elucidate pathogenic processes and an essential tool to test novel therapeutic strategies...

Descripción completa

Detalles Bibliográficos
Autores principales: Frank, Ines, Acharya, Arpan, Routhu, Nanda K., Aravantinou, Meropi, Harper, Justin L., Maldonado, Stephanie, Sole Cigoli, Maria, Semova, Stanka, Mazel, Svetlana, Paiardini, Mirko, Derby, Nina, Byrareddy, Siddappa N., Martinelli, Elena
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6700126/
https://www.ncbi.nlm.nih.gov/pubmed/31427605
http://dx.doi.org/10.1038/s41598-019-48354-3
_version_ 1783444802396749824
author Frank, Ines
Acharya, Arpan
Routhu, Nanda K.
Aravantinou, Meropi
Harper, Justin L.
Maldonado, Stephanie
Sole Cigoli, Maria
Semova, Stanka
Mazel, Svetlana
Paiardini, Mirko
Derby, Nina
Byrareddy, Siddappa N.
Martinelli, Elena
author_facet Frank, Ines
Acharya, Arpan
Routhu, Nanda K.
Aravantinou, Meropi
Harper, Justin L.
Maldonado, Stephanie
Sole Cigoli, Maria
Semova, Stanka
Mazel, Svetlana
Paiardini, Mirko
Derby, Nina
Byrareddy, Siddappa N.
Martinelli, Elena
author_sort Frank, Ines
collection PubMed
description The establishment of latent infection and poorly characterized viral reservoirs in tissues represent major obstacles to a definitive cure for HIV. Non-human primate (NHP) models of HIV infection are critical to elucidate pathogenic processes and an essential tool to test novel therapeutic strategies. Thus, the availability of novel assays to measure residual viral replication and reservoirs in NHP models may increase their utility in the search for an HIV cure. We developed a tat/rev induced limiting dilution assay to measure the frequency of CD4(+) T cells that express multiply-spliced(ms)_SIV RNA in presence and absence of stimulation. We validated the assay using cell lines and cells from blood and lymph nodes of SIV infected macaques. In vitro, SIV/SHIV TILDA detects only cells expressing viral proteins. In SIV/SHIV-infected macaques, CD4(+) T cells that express msSIV/SHIV RNA (TILDA data) were detected also in the setting of very low/undetectable viremia. TILDA data were significantly higher after stimulation and correlated with plasma viral load (pVL). Interestingly, TILDA data from early cART initiation correlated with peak and AUC pVL post-cART interruption. In summary, we developed an assay that may be useful in characterizing viral reservoirs and determining the effect of HIV interventions in NHP models.
format Online
Article
Text
id pubmed-6700126
institution National Center for Biotechnology Information
language English
publishDate 2019
publisher Nature Publishing Group UK
record_format MEDLINE/PubMed
spelling pubmed-67001262019-08-21 A Tat/Rev Induced Limiting Dilution Assay to Measure Viral Reservoirs in Non-Human Primate Models of HIV Infection Frank, Ines Acharya, Arpan Routhu, Nanda K. Aravantinou, Meropi Harper, Justin L. Maldonado, Stephanie Sole Cigoli, Maria Semova, Stanka Mazel, Svetlana Paiardini, Mirko Derby, Nina Byrareddy, Siddappa N. Martinelli, Elena Sci Rep Article The establishment of latent infection and poorly characterized viral reservoirs in tissues represent major obstacles to a definitive cure for HIV. Non-human primate (NHP) models of HIV infection are critical to elucidate pathogenic processes and an essential tool to test novel therapeutic strategies. Thus, the availability of novel assays to measure residual viral replication and reservoirs in NHP models may increase their utility in the search for an HIV cure. We developed a tat/rev induced limiting dilution assay to measure the frequency of CD4(+) T cells that express multiply-spliced(ms)_SIV RNA in presence and absence of stimulation. We validated the assay using cell lines and cells from blood and lymph nodes of SIV infected macaques. In vitro, SIV/SHIV TILDA detects only cells expressing viral proteins. In SIV/SHIV-infected macaques, CD4(+) T cells that express msSIV/SHIV RNA (TILDA data) were detected also in the setting of very low/undetectable viremia. TILDA data were significantly higher after stimulation and correlated with plasma viral load (pVL). Interestingly, TILDA data from early cART initiation correlated with peak and AUC pVL post-cART interruption. In summary, we developed an assay that may be useful in characterizing viral reservoirs and determining the effect of HIV interventions in NHP models. Nature Publishing Group UK 2019-08-19 /pmc/articles/PMC6700126/ /pubmed/31427605 http://dx.doi.org/10.1038/s41598-019-48354-3 Text en © The Author(s) 2019 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Frank, Ines
Acharya, Arpan
Routhu, Nanda K.
Aravantinou, Meropi
Harper, Justin L.
Maldonado, Stephanie
Sole Cigoli, Maria
Semova, Stanka
Mazel, Svetlana
Paiardini, Mirko
Derby, Nina
Byrareddy, Siddappa N.
Martinelli, Elena
A Tat/Rev Induced Limiting Dilution Assay to Measure Viral Reservoirs in Non-Human Primate Models of HIV Infection
title A Tat/Rev Induced Limiting Dilution Assay to Measure Viral Reservoirs in Non-Human Primate Models of HIV Infection
title_full A Tat/Rev Induced Limiting Dilution Assay to Measure Viral Reservoirs in Non-Human Primate Models of HIV Infection
title_fullStr A Tat/Rev Induced Limiting Dilution Assay to Measure Viral Reservoirs in Non-Human Primate Models of HIV Infection
title_full_unstemmed A Tat/Rev Induced Limiting Dilution Assay to Measure Viral Reservoirs in Non-Human Primate Models of HIV Infection
title_short A Tat/Rev Induced Limiting Dilution Assay to Measure Viral Reservoirs in Non-Human Primate Models of HIV Infection
title_sort tat/rev induced limiting dilution assay to measure viral reservoirs in non-human primate models of hiv infection
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6700126/
https://www.ncbi.nlm.nih.gov/pubmed/31427605
http://dx.doi.org/10.1038/s41598-019-48354-3
work_keys_str_mv AT frankines atatrevinducedlimitingdilutionassaytomeasureviralreservoirsinnonhumanprimatemodelsofhivinfection
AT acharyaarpan atatrevinducedlimitingdilutionassaytomeasureviralreservoirsinnonhumanprimatemodelsofhivinfection
AT routhunandak atatrevinducedlimitingdilutionassaytomeasureviralreservoirsinnonhumanprimatemodelsofhivinfection
AT aravantinoumeropi atatrevinducedlimitingdilutionassaytomeasureviralreservoirsinnonhumanprimatemodelsofhivinfection
AT harperjustinl atatrevinducedlimitingdilutionassaytomeasureviralreservoirsinnonhumanprimatemodelsofhivinfection
AT maldonadostephanie atatrevinducedlimitingdilutionassaytomeasureviralreservoirsinnonhumanprimatemodelsofhivinfection
AT solecigolimaria atatrevinducedlimitingdilutionassaytomeasureviralreservoirsinnonhumanprimatemodelsofhivinfection
AT semovastanka atatrevinducedlimitingdilutionassaytomeasureviralreservoirsinnonhumanprimatemodelsofhivinfection
AT mazelsvetlana atatrevinducedlimitingdilutionassaytomeasureviralreservoirsinnonhumanprimatemodelsofhivinfection
AT paiardinimirko atatrevinducedlimitingdilutionassaytomeasureviralreservoirsinnonhumanprimatemodelsofhivinfection
AT derbynina atatrevinducedlimitingdilutionassaytomeasureviralreservoirsinnonhumanprimatemodelsofhivinfection
AT byrareddysiddappan atatrevinducedlimitingdilutionassaytomeasureviralreservoirsinnonhumanprimatemodelsofhivinfection
AT martinellielena atatrevinducedlimitingdilutionassaytomeasureviralreservoirsinnonhumanprimatemodelsofhivinfection
AT frankines tatrevinducedlimitingdilutionassaytomeasureviralreservoirsinnonhumanprimatemodelsofhivinfection
AT acharyaarpan tatrevinducedlimitingdilutionassaytomeasureviralreservoirsinnonhumanprimatemodelsofhivinfection
AT routhunandak tatrevinducedlimitingdilutionassaytomeasureviralreservoirsinnonhumanprimatemodelsofhivinfection
AT aravantinoumeropi tatrevinducedlimitingdilutionassaytomeasureviralreservoirsinnonhumanprimatemodelsofhivinfection
AT harperjustinl tatrevinducedlimitingdilutionassaytomeasureviralreservoirsinnonhumanprimatemodelsofhivinfection
AT maldonadostephanie tatrevinducedlimitingdilutionassaytomeasureviralreservoirsinnonhumanprimatemodelsofhivinfection
AT solecigolimaria tatrevinducedlimitingdilutionassaytomeasureviralreservoirsinnonhumanprimatemodelsofhivinfection
AT semovastanka tatrevinducedlimitingdilutionassaytomeasureviralreservoirsinnonhumanprimatemodelsofhivinfection
AT mazelsvetlana tatrevinducedlimitingdilutionassaytomeasureviralreservoirsinnonhumanprimatemodelsofhivinfection
AT paiardinimirko tatrevinducedlimitingdilutionassaytomeasureviralreservoirsinnonhumanprimatemodelsofhivinfection
AT derbynina tatrevinducedlimitingdilutionassaytomeasureviralreservoirsinnonhumanprimatemodelsofhivinfection
AT byrareddysiddappan tatrevinducedlimitingdilutionassaytomeasureviralreservoirsinnonhumanprimatemodelsofhivinfection
AT martinellielena tatrevinducedlimitingdilutionassaytomeasureviralreservoirsinnonhumanprimatemodelsofhivinfection