High-throughput Characterization of HIV-1 Reservoir Reactivation Using a Single-Cell-in-Droplet PCR Assay
Reactivation of latent viral reservoirs is on the forefront of HIV-1 eradication research. However, it is unknown if latency reversing agents (LRAs) increase the level of viral transcription from cells producing HIV RNA or harboring transcriptionally-inactive (latent) infection. We therefore develop...
Ausführliche Beschreibung
Gespeichert in:
| Autor*in: |
Robert W. Yucha [verfasserIn] Kristen S. Hobbs [verfasserIn] Emily Hanhauser [verfasserIn] Louise E. Hogan [verfasserIn] Wildaliz Nieves [verfasserIn] Mehmet O. Ozen [verfasserIn] Fatih Inci [verfasserIn] Vanessa York [verfasserIn] Erica A. Gibson [verfasserIn] Cassandra Thanh [verfasserIn] Hadi Shafiee [verfasserIn] Rami El Assal [verfasserIn] Maja Kiselinova [verfasserIn] Yvonne P. Robles [verfasserIn] Helen Bae [verfasserIn] Kaitlyn S. Leadabrand [verfasserIn] ShuQi Wang [verfasserIn] Steven G. Deeks [verfasserIn] Daniel R. Kuritzkes [verfasserIn] Utkan Demirci [verfasserIn] Timothy J. Henrich [verfasserIn] |
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| Format: |
E-Artikel |
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| Sprache: |
Englisch |
| Erschienen: |
2017 |
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| Schlagwörter: |
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| Übergeordnetes Werk: |
In: EBioMedicine - Elsevier, 2015, 20(2017), C, Seite 217-229 |
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| Übergeordnetes Werk: |
volume:20 ; year:2017 ; number:C ; pages:217-229 |
| Links: |
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| DOI / URN: |
10.1016/j.ebiom.2017.05.006 |
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| Katalog-ID: |
DOAJ054412439 |
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| 100 | 0 | |a Robert W. Yucha |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a High-throughput Characterization of HIV-1 Reservoir Reactivation Using a Single-Cell-in-Droplet PCR Assay |
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| 520 | |a Reactivation of latent viral reservoirs is on the forefront of HIV-1 eradication research. However, it is unknown if latency reversing agents (LRAs) increase the level of viral transcription from cells producing HIV RNA or harboring transcriptionally-inactive (latent) infection. We therefore developed a microfluidic single-cell-in-droplet (scd)PCR assay to directly measure the number of CD4+ T cells that produce unspliced (us)RNA and multiply spliced (ms)RNA following ex vivo latency reversal with either an histone deacetylase inhibitor (romidepsin) or T cell receptor (TCR) stimulation. Detection of HIV-1 transcriptional activity can also be performed on hundreds of thousands of CD4+ T-cells in a single experiment. The scdPCR method was then applied to CD4+ T cells obtained from HIV-1-infected individuals on antiretroviral therapy. Overall, our results suggest that effects of LRAs on HIV-1 reactivation may be heterogeneous—increasing transcription from active cells in some cases and increasing the number of transcriptionally active cells in others. Genomic DNA and human mRNA isolated from HIV-1 reactivated cells could also be detected and quantified from individual cells. As a result, our assay has the potential to provide needed insight into various reservoir eradication strategies. | ||
| 650 | 4 | |a Human Immunodeficiency Virus (HIV) | |
| 650 | 4 | |a Single cell quantification | |
| 650 | 4 | |a Digital PCR | |
| 650 | 4 | |a HIV reservoirs | |
| 650 | 4 | |a HIV reactivation | |
| 650 | 4 | |a Histone deacetylase inhibitors | |
| 653 | 0 | |a Medicine | |
| 653 | 0 | |a R | |
| 653 | 0 | |a Medicine (General) | |
| 700 | 0 | |a Kristen S. Hobbs |e verfasserin |4 aut | |
| 700 | 0 | |a Emily Hanhauser |e verfasserin |4 aut | |
| 700 | 0 | |a Louise E. Hogan |e verfasserin |4 aut | |
| 700 | 0 | |a Wildaliz Nieves |e verfasserin |4 aut | |
| 700 | 0 | |a Mehmet O. Ozen |e verfasserin |4 aut | |
| 700 | 0 | |a Fatih Inci |e verfasserin |4 aut | |
| 700 | 0 | |a Vanessa York |e verfasserin |4 aut | |
| 700 | 0 | |a Erica A. Gibson |e verfasserin |4 aut | |
| 700 | 0 | |a Cassandra Thanh |e verfasserin |4 aut | |
| 700 | 0 | |a Hadi Shafiee |e verfasserin |4 aut | |
| 700 | 0 | |a Rami El Assal |e verfasserin |4 aut | |
| 700 | 0 | |a Maja Kiselinova |e verfasserin |4 aut | |
| 700 | 0 | |a Yvonne P. Robles |e verfasserin |4 aut | |
| 700 | 0 | |a Helen Bae |e verfasserin |4 aut | |
| 700 | 0 | |a Kaitlyn S. Leadabrand |e verfasserin |4 aut | |
| 700 | 0 | |a ShuQi Wang |e verfasserin |4 aut | |
| 700 | 0 | |a Steven G. Deeks |e verfasserin |4 aut | |
| 700 | 0 | |a Daniel R. Kuritzkes |e verfasserin |4 aut | |
| 700 | 0 | |a Utkan Demirci |e verfasserin |4 aut | |
| 700 | 0 | |a Timothy J. Henrich |e verfasserin |4 aut | |
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10.1016/j.ebiom.2017.05.006 doi (DE-627)DOAJ054412439 (DE-599)DOAJ7f06bfd56eae4628b883889050466cda DE-627 ger DE-627 rakwb eng R5-920 Robert W. Yucha verfasserin aut High-throughput Characterization of HIV-1 Reservoir Reactivation Using a Single-Cell-in-Droplet PCR Assay 2017 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Reactivation of latent viral reservoirs is on the forefront of HIV-1 eradication research. However, it is unknown if latency reversing agents (LRAs) increase the level of viral transcription from cells producing HIV RNA or harboring transcriptionally-inactive (latent) infection. We therefore developed a microfluidic single-cell-in-droplet (scd)PCR assay to directly measure the number of CD4+ T cells that produce unspliced (us)RNA and multiply spliced (ms)RNA following ex vivo latency reversal with either an histone deacetylase inhibitor (romidepsin) or T cell receptor (TCR) stimulation. Detection of HIV-1 transcriptional activity can also be performed on hundreds of thousands of CD4+ T-cells in a single experiment. The scdPCR method was then applied to CD4+ T cells obtained from HIV-1-infected individuals on antiretroviral therapy. Overall, our results suggest that effects of LRAs on HIV-1 reactivation may be heterogeneous—increasing transcription from active cells in some cases and increasing the number of transcriptionally active cells in others. Genomic DNA and human mRNA isolated from HIV-1 reactivated cells could also be detected and quantified from individual cells. As a result, our assay has the potential to provide needed insight into various reservoir eradication strategies. Human Immunodeficiency Virus (HIV) Single cell quantification Digital PCR HIV reservoirs HIV reactivation Histone deacetylase inhibitors Medicine R Medicine (General) Kristen S. Hobbs verfasserin aut Emily Hanhauser verfasserin aut Louise E. Hogan verfasserin aut Wildaliz Nieves verfasserin aut Mehmet O. Ozen verfasserin aut Fatih Inci verfasserin aut Vanessa York verfasserin aut Erica A. Gibson verfasserin aut Cassandra Thanh verfasserin aut Hadi Shafiee verfasserin aut Rami El Assal verfasserin aut Maja Kiselinova verfasserin aut Yvonne P. Robles verfasserin aut Helen Bae verfasserin aut Kaitlyn S. Leadabrand verfasserin aut ShuQi Wang verfasserin aut Steven G. Deeks verfasserin aut Daniel R. Kuritzkes verfasserin aut Utkan Demirci verfasserin aut Timothy J. Henrich verfasserin aut In EBioMedicine Elsevier, 2015 20(2017), C, Seite 217-229 (DE-627)802540074 (DE-600)2799017-5 23523964 nnns volume:20 year:2017 number:C pages:217-229 https://doi.org/10.1016/j.ebiom.2017.05.006 kostenfrei https://doaj.org/article/7f06bfd56eae4628b883889050466cda kostenfrei http://www.sciencedirect.com/science/article/pii/S2352396417301974 kostenfrei https://doaj.org/toc/2352-3964 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2007 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2026 GBV_ILN_2027 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2049 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2059 GBV_ILN_2061 GBV_ILN_2064 GBV_ILN_2068 GBV_ILN_2088 GBV_ILN_2110 GBV_ILN_2112 GBV_ILN_2129 GBV_ILN_2143 GBV_ILN_2153 GBV_ILN_2190 GBV_ILN_2336 GBV_ILN_2470 GBV_ILN_2507 GBV_ILN_4012 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4326 GBV_ILN_4333 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4393 GBV_ILN_4700 AR 20 2017 C 217-229 |
| spelling |
10.1016/j.ebiom.2017.05.006 doi (DE-627)DOAJ054412439 (DE-599)DOAJ7f06bfd56eae4628b883889050466cda DE-627 ger DE-627 rakwb eng R5-920 Robert W. Yucha verfasserin aut High-throughput Characterization of HIV-1 Reservoir Reactivation Using a Single-Cell-in-Droplet PCR Assay 2017 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Reactivation of latent viral reservoirs is on the forefront of HIV-1 eradication research. However, it is unknown if latency reversing agents (LRAs) increase the level of viral transcription from cells producing HIV RNA or harboring transcriptionally-inactive (latent) infection. We therefore developed a microfluidic single-cell-in-droplet (scd)PCR assay to directly measure the number of CD4+ T cells that produce unspliced (us)RNA and multiply spliced (ms)RNA following ex vivo latency reversal with either an histone deacetylase inhibitor (romidepsin) or T cell receptor (TCR) stimulation. Detection of HIV-1 transcriptional activity can also be performed on hundreds of thousands of CD4+ T-cells in a single experiment. The scdPCR method was then applied to CD4+ T cells obtained from HIV-1-infected individuals on antiretroviral therapy. Overall, our results suggest that effects of LRAs on HIV-1 reactivation may be heterogeneous—increasing transcription from active cells in some cases and increasing the number of transcriptionally active cells in others. Genomic DNA and human mRNA isolated from HIV-1 reactivated cells could also be detected and quantified from individual cells. As a result, our assay has the potential to provide needed insight into various reservoir eradication strategies. Human Immunodeficiency Virus (HIV) Single cell quantification Digital PCR HIV reservoirs HIV reactivation Histone deacetylase inhibitors Medicine R Medicine (General) Kristen S. Hobbs verfasserin aut Emily Hanhauser verfasserin aut Louise E. Hogan verfasserin aut Wildaliz Nieves verfasserin aut Mehmet O. Ozen verfasserin aut Fatih Inci verfasserin aut Vanessa York verfasserin aut Erica A. Gibson verfasserin aut Cassandra Thanh verfasserin aut Hadi Shafiee verfasserin aut Rami El Assal verfasserin aut Maja Kiselinova verfasserin aut Yvonne P. Robles verfasserin aut Helen Bae verfasserin aut Kaitlyn S. Leadabrand verfasserin aut ShuQi Wang verfasserin aut Steven G. Deeks verfasserin aut Daniel R. Kuritzkes verfasserin aut Utkan Demirci verfasserin aut Timothy J. Henrich verfasserin aut In EBioMedicine Elsevier, 2015 20(2017), C, Seite 217-229 (DE-627)802540074 (DE-600)2799017-5 23523964 nnns volume:20 year:2017 number:C pages:217-229 https://doi.org/10.1016/j.ebiom.2017.05.006 kostenfrei https://doaj.org/article/7f06bfd56eae4628b883889050466cda kostenfrei http://www.sciencedirect.com/science/article/pii/S2352396417301974 kostenfrei https://doaj.org/toc/2352-3964 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2007 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2026 GBV_ILN_2027 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2049 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2059 GBV_ILN_2061 GBV_ILN_2064 GBV_ILN_2068 GBV_ILN_2088 GBV_ILN_2110 GBV_ILN_2112 GBV_ILN_2129 GBV_ILN_2143 GBV_ILN_2153 GBV_ILN_2190 GBV_ILN_2336 GBV_ILN_2470 GBV_ILN_2507 GBV_ILN_4012 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4326 GBV_ILN_4333 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4393 GBV_ILN_4700 AR 20 2017 C 217-229 |
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10.1016/j.ebiom.2017.05.006 doi (DE-627)DOAJ054412439 (DE-599)DOAJ7f06bfd56eae4628b883889050466cda DE-627 ger DE-627 rakwb eng R5-920 Robert W. Yucha verfasserin aut High-throughput Characterization of HIV-1 Reservoir Reactivation Using a Single-Cell-in-Droplet PCR Assay 2017 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Reactivation of latent viral reservoirs is on the forefront of HIV-1 eradication research. However, it is unknown if latency reversing agents (LRAs) increase the level of viral transcription from cells producing HIV RNA or harboring transcriptionally-inactive (latent) infection. We therefore developed a microfluidic single-cell-in-droplet (scd)PCR assay to directly measure the number of CD4+ T cells that produce unspliced (us)RNA and multiply spliced (ms)RNA following ex vivo latency reversal with either an histone deacetylase inhibitor (romidepsin) or T cell receptor (TCR) stimulation. Detection of HIV-1 transcriptional activity can also be performed on hundreds of thousands of CD4+ T-cells in a single experiment. The scdPCR method was then applied to CD4+ T cells obtained from HIV-1-infected individuals on antiretroviral therapy. Overall, our results suggest that effects of LRAs on HIV-1 reactivation may be heterogeneous—increasing transcription from active cells in some cases and increasing the number of transcriptionally active cells in others. Genomic DNA and human mRNA isolated from HIV-1 reactivated cells could also be detected and quantified from individual cells. As a result, our assay has the potential to provide needed insight into various reservoir eradication strategies. Human Immunodeficiency Virus (HIV) Single cell quantification Digital PCR HIV reservoirs HIV reactivation Histone deacetylase inhibitors Medicine R Medicine (General) Kristen S. Hobbs verfasserin aut Emily Hanhauser verfasserin aut Louise E. Hogan verfasserin aut Wildaliz Nieves verfasserin aut Mehmet O. Ozen verfasserin aut Fatih Inci verfasserin aut Vanessa York verfasserin aut Erica A. Gibson verfasserin aut Cassandra Thanh verfasserin aut Hadi Shafiee verfasserin aut Rami El Assal verfasserin aut Maja Kiselinova verfasserin aut Yvonne P. Robles verfasserin aut Helen Bae verfasserin aut Kaitlyn S. Leadabrand verfasserin aut ShuQi Wang verfasserin aut Steven G. Deeks verfasserin aut Daniel R. Kuritzkes verfasserin aut Utkan Demirci verfasserin aut Timothy J. Henrich verfasserin aut In EBioMedicine Elsevier, 2015 20(2017), C, Seite 217-229 (DE-627)802540074 (DE-600)2799017-5 23523964 nnns volume:20 year:2017 number:C pages:217-229 https://doi.org/10.1016/j.ebiom.2017.05.006 kostenfrei https://doaj.org/article/7f06bfd56eae4628b883889050466cda kostenfrei http://www.sciencedirect.com/science/article/pii/S2352396417301974 kostenfrei https://doaj.org/toc/2352-3964 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2007 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2026 GBV_ILN_2027 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2049 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2059 GBV_ILN_2061 GBV_ILN_2064 GBV_ILN_2068 GBV_ILN_2088 GBV_ILN_2110 GBV_ILN_2112 GBV_ILN_2129 GBV_ILN_2143 GBV_ILN_2153 GBV_ILN_2190 GBV_ILN_2336 GBV_ILN_2470 GBV_ILN_2507 GBV_ILN_4012 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4326 GBV_ILN_4333 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4393 GBV_ILN_4700 AR 20 2017 C 217-229 |
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10.1016/j.ebiom.2017.05.006 doi (DE-627)DOAJ054412439 (DE-599)DOAJ7f06bfd56eae4628b883889050466cda DE-627 ger DE-627 rakwb eng R5-920 Robert W. Yucha verfasserin aut High-throughput Characterization of HIV-1 Reservoir Reactivation Using a Single-Cell-in-Droplet PCR Assay 2017 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Reactivation of latent viral reservoirs is on the forefront of HIV-1 eradication research. However, it is unknown if latency reversing agents (LRAs) increase the level of viral transcription from cells producing HIV RNA or harboring transcriptionally-inactive (latent) infection. We therefore developed a microfluidic single-cell-in-droplet (scd)PCR assay to directly measure the number of CD4+ T cells that produce unspliced (us)RNA and multiply spliced (ms)RNA following ex vivo latency reversal with either an histone deacetylase inhibitor (romidepsin) or T cell receptor (TCR) stimulation. Detection of HIV-1 transcriptional activity can also be performed on hundreds of thousands of CD4+ T-cells in a single experiment. The scdPCR method was then applied to CD4+ T cells obtained from HIV-1-infected individuals on antiretroviral therapy. Overall, our results suggest that effects of LRAs on HIV-1 reactivation may be heterogeneous—increasing transcription from active cells in some cases and increasing the number of transcriptionally active cells in others. Genomic DNA and human mRNA isolated from HIV-1 reactivated cells could also be detected and quantified from individual cells. As a result, our assay has the potential to provide needed insight into various reservoir eradication strategies. Human Immunodeficiency Virus (HIV) Single cell quantification Digital PCR HIV reservoirs HIV reactivation Histone deacetylase inhibitors Medicine R Medicine (General) Kristen S. Hobbs verfasserin aut Emily Hanhauser verfasserin aut Louise E. Hogan verfasserin aut Wildaliz Nieves verfasserin aut Mehmet O. Ozen verfasserin aut Fatih Inci verfasserin aut Vanessa York verfasserin aut Erica A. Gibson verfasserin aut Cassandra Thanh verfasserin aut Hadi Shafiee verfasserin aut Rami El Assal verfasserin aut Maja Kiselinova verfasserin aut Yvonne P. Robles verfasserin aut Helen Bae verfasserin aut Kaitlyn S. Leadabrand verfasserin aut ShuQi Wang verfasserin aut Steven G. Deeks verfasserin aut Daniel R. Kuritzkes verfasserin aut Utkan Demirci verfasserin aut Timothy J. Henrich verfasserin aut In EBioMedicine Elsevier, 2015 20(2017), C, Seite 217-229 (DE-627)802540074 (DE-600)2799017-5 23523964 nnns volume:20 year:2017 number:C pages:217-229 https://doi.org/10.1016/j.ebiom.2017.05.006 kostenfrei https://doaj.org/article/7f06bfd56eae4628b883889050466cda kostenfrei http://www.sciencedirect.com/science/article/pii/S2352396417301974 kostenfrei https://doaj.org/toc/2352-3964 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2007 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2026 GBV_ILN_2027 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2049 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2059 GBV_ILN_2061 GBV_ILN_2064 GBV_ILN_2068 GBV_ILN_2088 GBV_ILN_2110 GBV_ILN_2112 GBV_ILN_2129 GBV_ILN_2143 GBV_ILN_2153 GBV_ILN_2190 GBV_ILN_2336 GBV_ILN_2470 GBV_ILN_2507 GBV_ILN_4012 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4326 GBV_ILN_4333 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4393 GBV_ILN_4700 AR 20 2017 C 217-229 |
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10.1016/j.ebiom.2017.05.006 doi (DE-627)DOAJ054412439 (DE-599)DOAJ7f06bfd56eae4628b883889050466cda DE-627 ger DE-627 rakwb eng R5-920 Robert W. Yucha verfasserin aut High-throughput Characterization of HIV-1 Reservoir Reactivation Using a Single-Cell-in-Droplet PCR Assay 2017 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Reactivation of latent viral reservoirs is on the forefront of HIV-1 eradication research. However, it is unknown if latency reversing agents (LRAs) increase the level of viral transcription from cells producing HIV RNA or harboring transcriptionally-inactive (latent) infection. We therefore developed a microfluidic single-cell-in-droplet (scd)PCR assay to directly measure the number of CD4+ T cells that produce unspliced (us)RNA and multiply spliced (ms)RNA following ex vivo latency reversal with either an histone deacetylase inhibitor (romidepsin) or T cell receptor (TCR) stimulation. Detection of HIV-1 transcriptional activity can also be performed on hundreds of thousands of CD4+ T-cells in a single experiment. The scdPCR method was then applied to CD4+ T cells obtained from HIV-1-infected individuals on antiretroviral therapy. Overall, our results suggest that effects of LRAs on HIV-1 reactivation may be heterogeneous—increasing transcription from active cells in some cases and increasing the number of transcriptionally active cells in others. Genomic DNA and human mRNA isolated from HIV-1 reactivated cells could also be detected and quantified from individual cells. As a result, our assay has the potential to provide needed insight into various reservoir eradication strategies. Human Immunodeficiency Virus (HIV) Single cell quantification Digital PCR HIV reservoirs HIV reactivation Histone deacetylase inhibitors Medicine R Medicine (General) Kristen S. Hobbs verfasserin aut Emily Hanhauser verfasserin aut Louise E. Hogan verfasserin aut Wildaliz Nieves verfasserin aut Mehmet O. Ozen verfasserin aut Fatih Inci verfasserin aut Vanessa York verfasserin aut Erica A. Gibson verfasserin aut Cassandra Thanh verfasserin aut Hadi Shafiee verfasserin aut Rami El Assal verfasserin aut Maja Kiselinova verfasserin aut Yvonne P. Robles verfasserin aut Helen Bae verfasserin aut Kaitlyn S. Leadabrand verfasserin aut ShuQi Wang verfasserin aut Steven G. Deeks verfasserin aut Daniel R. Kuritzkes verfasserin aut Utkan Demirci verfasserin aut Timothy J. Henrich verfasserin aut In EBioMedicine Elsevier, 2015 20(2017), C, Seite 217-229 (DE-627)802540074 (DE-600)2799017-5 23523964 nnns volume:20 year:2017 number:C pages:217-229 https://doi.org/10.1016/j.ebiom.2017.05.006 kostenfrei https://doaj.org/article/7f06bfd56eae4628b883889050466cda kostenfrei http://www.sciencedirect.com/science/article/pii/S2352396417301974 kostenfrei https://doaj.org/toc/2352-3964 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2007 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2026 GBV_ILN_2027 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2049 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2059 GBV_ILN_2061 GBV_ILN_2064 GBV_ILN_2068 GBV_ILN_2088 GBV_ILN_2110 GBV_ILN_2112 GBV_ILN_2129 GBV_ILN_2143 GBV_ILN_2153 GBV_ILN_2190 GBV_ILN_2336 GBV_ILN_2470 GBV_ILN_2507 GBV_ILN_4012 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4326 GBV_ILN_4333 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4393 GBV_ILN_4700 AR 20 2017 C 217-229 |
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Robert W. Yucha @@aut@@ Kristen S. Hobbs @@aut@@ Emily Hanhauser @@aut@@ Louise E. Hogan @@aut@@ Wildaliz Nieves @@aut@@ Mehmet O. Ozen @@aut@@ Fatih Inci @@aut@@ Vanessa York @@aut@@ Erica A. Gibson @@aut@@ Cassandra Thanh @@aut@@ Hadi Shafiee @@aut@@ Rami El Assal @@aut@@ Maja Kiselinova @@aut@@ Yvonne P. Robles @@aut@@ Helen Bae @@aut@@ Kaitlyn S. Leadabrand @@aut@@ ShuQi Wang @@aut@@ Steven G. Deeks @@aut@@ Daniel R. Kuritzkes @@aut@@ Utkan Demirci @@aut@@ Timothy J. Henrich @@aut@@ |
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Robert W. Yucha |
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Robert W. Yucha misc R5-920 misc Human Immunodeficiency Virus (HIV) misc Single cell quantification misc Digital PCR misc HIV reservoirs misc HIV reactivation misc Histone deacetylase inhibitors misc Medicine misc R misc Medicine (General) High-throughput Characterization of HIV-1 Reservoir Reactivation Using a Single-Cell-in-Droplet PCR Assay |
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R5-920 High-throughput Characterization of HIV-1 Reservoir Reactivation Using a Single-Cell-in-Droplet PCR Assay Human Immunodeficiency Virus (HIV) Single cell quantification Digital PCR HIV reservoirs HIV reactivation Histone deacetylase inhibitors |
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Robert W. Yucha Kristen S. Hobbs Emily Hanhauser Louise E. Hogan Wildaliz Nieves Mehmet O. Ozen Fatih Inci Vanessa York Erica A. Gibson Cassandra Thanh Hadi Shafiee Rami El Assal Maja Kiselinova Yvonne P. Robles Helen Bae Kaitlyn S. Leadabrand ShuQi Wang Steven G. Deeks Daniel R. Kuritzkes Utkan Demirci Timothy J. Henrich |
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high-throughput characterization of hiv-1 reservoir reactivation using a single-cell-in-droplet pcr assay |
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High-throughput Characterization of HIV-1 Reservoir Reactivation Using a Single-Cell-in-Droplet PCR Assay |
| abstract |
Reactivation of latent viral reservoirs is on the forefront of HIV-1 eradication research. However, it is unknown if latency reversing agents (LRAs) increase the level of viral transcription from cells producing HIV RNA or harboring transcriptionally-inactive (latent) infection. We therefore developed a microfluidic single-cell-in-droplet (scd)PCR assay to directly measure the number of CD4+ T cells that produce unspliced (us)RNA and multiply spliced (ms)RNA following ex vivo latency reversal with either an histone deacetylase inhibitor (romidepsin) or T cell receptor (TCR) stimulation. Detection of HIV-1 transcriptional activity can also be performed on hundreds of thousands of CD4+ T-cells in a single experiment. The scdPCR method was then applied to CD4+ T cells obtained from HIV-1-infected individuals on antiretroviral therapy. Overall, our results suggest that effects of LRAs on HIV-1 reactivation may be heterogeneous—increasing transcription from active cells in some cases and increasing the number of transcriptionally active cells in others. Genomic DNA and human mRNA isolated from HIV-1 reactivated cells could also be detected and quantified from individual cells. As a result, our assay has the potential to provide needed insight into various reservoir eradication strategies. |
| abstractGer |
Reactivation of latent viral reservoirs is on the forefront of HIV-1 eradication research. However, it is unknown if latency reversing agents (LRAs) increase the level of viral transcription from cells producing HIV RNA or harboring transcriptionally-inactive (latent) infection. We therefore developed a microfluidic single-cell-in-droplet (scd)PCR assay to directly measure the number of CD4+ T cells that produce unspliced (us)RNA and multiply spliced (ms)RNA following ex vivo latency reversal with either an histone deacetylase inhibitor (romidepsin) or T cell receptor (TCR) stimulation. Detection of HIV-1 transcriptional activity can also be performed on hundreds of thousands of CD4+ T-cells in a single experiment. The scdPCR method was then applied to CD4+ T cells obtained from HIV-1-infected individuals on antiretroviral therapy. Overall, our results suggest that effects of LRAs on HIV-1 reactivation may be heterogeneous—increasing transcription from active cells in some cases and increasing the number of transcriptionally active cells in others. Genomic DNA and human mRNA isolated from HIV-1 reactivated cells could also be detected and quantified from individual cells. As a result, our assay has the potential to provide needed insight into various reservoir eradication strategies. |
| abstract_unstemmed |
Reactivation of latent viral reservoirs is on the forefront of HIV-1 eradication research. However, it is unknown if latency reversing agents (LRAs) increase the level of viral transcription from cells producing HIV RNA or harboring transcriptionally-inactive (latent) infection. We therefore developed a microfluidic single-cell-in-droplet (scd)PCR assay to directly measure the number of CD4+ T cells that produce unspliced (us)RNA and multiply spliced (ms)RNA following ex vivo latency reversal with either an histone deacetylase inhibitor (romidepsin) or T cell receptor (TCR) stimulation. Detection of HIV-1 transcriptional activity can also be performed on hundreds of thousands of CD4+ T-cells in a single experiment. The scdPCR method was then applied to CD4+ T cells obtained from HIV-1-infected individuals on antiretroviral therapy. Overall, our results suggest that effects of LRAs on HIV-1 reactivation may be heterogeneous—increasing transcription from active cells in some cases and increasing the number of transcriptionally active cells in others. Genomic DNA and human mRNA isolated from HIV-1 reactivated cells could also be detected and quantified from individual cells. As a result, our assay has the potential to provide needed insight into various reservoir eradication strategies. |
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