Retinoic Acid Improves the Recovery of Replication-Competent Virus from Latent SIV Infected Cells
The accurate estimation and eradication of Human Immunodeficiency Virus (HIV) viral reservoirs is limited by the incomplete reactivation of cells harboring the latent replication-competent virus. We investigated whether the in vitro and in vivo addition of retinoic acid (RA) enhances virus replicati...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2020-09-01
|
| Series: | Cells |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2073-4409/9/9/2076 |
| _version_ | 1827706436911628288 |
|---|---|
| author | Omalla A. Olwenyi Arpan Acharya Nanda Kishore Routhu Keely Pierzchalski Jace W. Jones Maureen A. Kane Neil Sidell Mahesh Mohan Siddappa N. Byrareddy |
| author_facet | Omalla A. Olwenyi Arpan Acharya Nanda Kishore Routhu Keely Pierzchalski Jace W. Jones Maureen A. Kane Neil Sidell Mahesh Mohan Siddappa N. Byrareddy |
| author_sort | Omalla A. Olwenyi |
| collection | DOAJ |
| description | The accurate estimation and eradication of Human Immunodeficiency Virus (HIV) viral reservoirs is limited by the incomplete reactivation of cells harboring the latent replication-competent virus. We investigated whether the in vitro and in vivo addition of retinoic acid (RA) enhances virus replication and improves the detection of latent virus. Peripheral blood mononuclear cells (PBMCs) from naive and anti-retroviral therapy (ART)-treated SIV-infected rhesus macaques (RMs) were cultured in vitro with anti-CD3/CD28 + IL-2 in the presence/absence of RA. Viral RNA and p27 levels were quantified using RT-qPCR and ELISA, respectively. Viral reservoirs were estimated using the Tat/Rev-Induced Limited Dilution Assay (TILDA) and Quantitative Viral Outgrowth Assay (QVOA). In vitro and in vivo measures revealed that there was also an increase in viral replication in RA-treated versus without RA conditions. In parallel, the addition of RA to either CD3/CD28 or phorbol myristate acetate (PMA)/ionomycin during QVOA and TILDA, respectively, was shown to augment reactivation of the replication-competent viral reservoir in anti-retroviral therapy (ART)-suppressed RMs as shown by a greater than 2.3-fold increase for QVOA and 1 to 2-fold increments for multi-spliced RNA per million CD4<sup>+</sup> T cells. The use of RA can be a useful approach to enhance the efficiency of current protocols used for in vitro and potentially in vivo estimates of CD4<sup>+</sup> T cell latent reservoirs. In addition, flow cytometry analysis revealed that RA improved estimates of various viral reservoir assays by eliciting broad CD4 T-cell activation as demonstrated by elevated CD25 and CD38 but reduced CD69 and PD-1 expressing cells. |
| first_indexed | 2024-03-10T16:24:21Z |
| format | Article |
| id | doaj.art-4dcf15b9930a4d39ae3e2c00fca3efc0 |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-10T16:24:21Z |
| publishDate | 2020-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj.art-4dcf15b9930a4d39ae3e2c00fca3efc02023-11-20T13:21:11ZengMDPI AGCells2073-44092020-09-0199207610.3390/cells9092076Retinoic Acid Improves the Recovery of Replication-Competent Virus from Latent SIV Infected CellsOmalla A. Olwenyi0Arpan Acharya1Nanda Kishore Routhu2Keely Pierzchalski3Jace W. Jones4Maureen A. Kane5Neil Sidell6Mahesh Mohan7Siddappa N. Byrareddy8Department of Pharmacology and Experimental Neuroscience, University of Nebraska Medical Center, Omaha, NE 68198-5800, USADepartment of Pharmacology and Experimental Neuroscience, University of Nebraska Medical Center, Omaha, NE 68198-5800, USADepartment of Pharmacology and Experimental Neuroscience, University of Nebraska Medical Center, Omaha, NE 68198-5800, USADepartment of Pharmaceutical Sciences, University of Maryland School of Pharmacy, Baltimore, MD 21201, USADepartment of Pharmaceutical Sciences, University of Maryland School of Pharmacy, Baltimore, MD 21201, USADepartment of Pharmaceutical Sciences, University of Maryland School of Pharmacy, Baltimore, MD 21201, USADepartment of Obstetrics and Gynecology, Emory University School of Medicine, Atlanta, GA 30322, USATexas Biomedical Research Institute, Southwest National Primate Research Institute, San Antonio, TX 78227, USADepartment of Pharmacology and Experimental Neuroscience, University of Nebraska Medical Center, Omaha, NE 68198-5800, USAThe accurate estimation and eradication of Human Immunodeficiency Virus (HIV) viral reservoirs is limited by the incomplete reactivation of cells harboring the latent replication-competent virus. We investigated whether the in vitro and in vivo addition of retinoic acid (RA) enhances virus replication and improves the detection of latent virus. Peripheral blood mononuclear cells (PBMCs) from naive and anti-retroviral therapy (ART)-treated SIV-infected rhesus macaques (RMs) were cultured in vitro with anti-CD3/CD28 + IL-2 in the presence/absence of RA. Viral RNA and p27 levels were quantified using RT-qPCR and ELISA, respectively. Viral reservoirs were estimated using the Tat/Rev-Induced Limited Dilution Assay (TILDA) and Quantitative Viral Outgrowth Assay (QVOA). In vitro and in vivo measures revealed that there was also an increase in viral replication in RA-treated versus without RA conditions. In parallel, the addition of RA to either CD3/CD28 or phorbol myristate acetate (PMA)/ionomycin during QVOA and TILDA, respectively, was shown to augment reactivation of the replication-competent viral reservoir in anti-retroviral therapy (ART)-suppressed RMs as shown by a greater than 2.3-fold increase for QVOA and 1 to 2-fold increments for multi-spliced RNA per million CD4<sup>+</sup> T cells. The use of RA can be a useful approach to enhance the efficiency of current protocols used for in vitro and potentially in vivo estimates of CD4<sup>+</sup> T cell latent reservoirs. In addition, flow cytometry analysis revealed that RA improved estimates of various viral reservoir assays by eliciting broad CD4 T-cell activation as demonstrated by elevated CD25 and CD38 but reduced CD69 and PD-1 expressing cells.https://www.mdpi.com/2073-4409/9/9/2076immune activationretinoic acidCD4 T cellsPBMCsSIVviral load and provirus |
| spellingShingle | Omalla A. Olwenyi Arpan Acharya Nanda Kishore Routhu Keely Pierzchalski Jace W. Jones Maureen A. Kane Neil Sidell Mahesh Mohan Siddappa N. Byrareddy Retinoic Acid Improves the Recovery of Replication-Competent Virus from Latent SIV Infected Cells Cells immune activation retinoic acid CD4 T cells PBMCs SIV viral load and provirus |
| title | Retinoic Acid Improves the Recovery of Replication-Competent Virus from Latent SIV Infected Cells |
| title_full | Retinoic Acid Improves the Recovery of Replication-Competent Virus from Latent SIV Infected Cells |
| title_fullStr | Retinoic Acid Improves the Recovery of Replication-Competent Virus from Latent SIV Infected Cells |
| title_full_unstemmed | Retinoic Acid Improves the Recovery of Replication-Competent Virus from Latent SIV Infected Cells |
| title_short | Retinoic Acid Improves the Recovery of Replication-Competent Virus from Latent SIV Infected Cells |
| title_sort | retinoic acid improves the recovery of replication competent virus from latent siv infected cells |
| topic | immune activation retinoic acid CD4 T cells PBMCs SIV viral load and provirus |
| url | https://www.mdpi.com/2073-4409/9/9/2076 |
| work_keys_str_mv | AT omallaaolwenyi retinoicacidimprovestherecoveryofreplicationcompetentvirusfromlatentsivinfectedcells AT arpanacharya retinoicacidimprovestherecoveryofreplicationcompetentvirusfromlatentsivinfectedcells AT nandakishorerouthu retinoicacidimprovestherecoveryofreplicationcompetentvirusfromlatentsivinfectedcells AT keelypierzchalski retinoicacidimprovestherecoveryofreplicationcompetentvirusfromlatentsivinfectedcells AT jacewjones retinoicacidimprovestherecoveryofreplicationcompetentvirusfromlatentsivinfectedcells AT maureenakane retinoicacidimprovestherecoveryofreplicationcompetentvirusfromlatentsivinfectedcells AT neilsidell retinoicacidimprovestherecoveryofreplicationcompetentvirusfromlatentsivinfectedcells AT maheshmohan retinoicacidimprovestherecoveryofreplicationcompetentvirusfromlatentsivinfectedcells AT siddappanbyrareddy retinoicacidimprovestherecoveryofreplicationcompetentvirusfromlatentsivinfectedcells |