SpringerLink
Log in

An Ultrasensitive p24 Assay to Measure HIV-1 in Diverse Biological Matrixes

  • Protocol
  • First Online:
HIV Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2807))

  • 300 Accesses

Abstract

Assays to study HIV persistence are crucial to evaluate therapeutic strategies aimed toward an HIV cure. Several assays have been developed to date that rely on the measurement of nucleic acids. In recent years, the advancement of ultrasensitive technologies for the detection of proteins has improved our understanding of the role of translation-competent reservoirs in HIV persistence. In this chapter, we describe the development of an ultrasensitive p24 ELISA that uses planar array technology. This assay allows for the detection of HIV-1 p24 in the low fg/ml range in different biological matrixes, including cell lysates. This assay can be used to investigate the efficacy of latency reversing agents to reactivate HIV or to evaluate the persistence of translation-competent reservoirs in people living with HIV (PWH) in cells or diverse biological fluids.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Protocol
EUR 44.95
Price includes VAT (Germany)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
EUR 181.89
Price includes VAT (Germany)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Hardcover Book
EUR 235.39
Price includes VAT (Germany)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free ship** worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Abdel-Mohsen M, Richman D, Siliciano RF, Nussenzweig MC, Howell BJ, Martinez-Picado J, Chomont N, Bar KJ, Yu XG, Lichterfeld M, Alcami J, Hazuda D, Bushman F, Siliciano JD, Betts MR, Spivak AM, Planelles V, Hahn BH, Smith DM, Ho YC, Buzon MJ, Gaebler C, Paiardini M, Li Q, Estes JD, Hope TJ, Kostman J, Mounzer K, Caskey M, Fox L, Frank I, Riley JL, Tebas P, Montaner LJ, infection B-HDCtCH (2020) Recommendations for measuring HIV reservoir size in cure-directed clinical trials. Nat Med 26(9):1339–1350. https://doi.org/10.1038/s41591-020-1022-1

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Procopio FA, Fromentin R, Kulpa DA, Brehm JH, Bebin AG, Strain MC, Richman DD, O’Doherty U, Palmer S, Hecht FM, Hoh R, Barnard RJ, Miller MD, Hazuda DJ, Deeks SG, Sekaly RP, Chomont N (2015) A novel assay to measure the magnitude of the inducible viral reservoir in HIV-infected individuals. EBioMedicine 2(8):874–883. https://doi.org/10.1016/j.ebiom.2015.06.019

    Article  PubMed  PubMed Central  Google Scholar 

  3. Butler SL, Hansen MS, Bushman FD (2001) A quantitative assay for HIV DNA integration in vivo. Nat Med 7(5):631–634. https://doi.org/10.1038/87979

    Article  CAS  PubMed  Google Scholar 

  4. O’Doherty U, Swiggard WJ, Jeyakumar D, McGain D, Malim MH (2002) A sensitive, quantitative assay for human immunodeficiency virus type 1 integration. J Virol 76(21):10942–10950

    Article  PubMed  PubMed Central  Google Scholar 

  5. Bruner KM, Wang Z, Simonetti FR, Bender AM, Kwon KJ, Sengupta S, Fray EJ, Beg SA, Antar AAR, Jenike KM, Bertagnolli LN, Capoferri AA, Kufera JT, Timmons A, Nobles C, Gregg J, Wada N, Ho YC, Zhang H, Margolick JB, Blankson JN, Deeks SG, Bushman FD, Siliciano JD, Laird GM, Siliciano RF (2019) A quantitative approach for measuring the reservoir of latent HIV-1 proviruses. Nature 566(7742):120–125. https://doi.org/10.1038/s41586-019-0898-8

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Yukl SA, Kaiser P, Kim P, Telwatte S, Joshi SK, Vu M, Lampiris H, Wong JK (2018) HIV latency in isolated patient CD4(+) T cells may be due to blocks in HIV transcriptional elongation, completion, and splicing. Sci Transl Med 10(430). https://doi.org/10.1126/scitranslmed.aap9927

  7. Pardons M, Baxter AE, Massanella M, Pagliuzza A, Fromentin R, Dufour C, Leyre L, Routy JP, Kaufmann DE, Chomont N (2019) Single-cell characterization and quantification of translation-competent viral reservoirs in treated and untreated HIV infection. PLoS Pathog 15(2):e1007619. https://doi.org/10.1371/journal.ppat.1007619

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Grau-Exposito J, Luque-Ballesteros L, Navarro J, Curran A, Burgos J, Ribera E, Torrella A, Planas B, Badia R, Martin-Castillo M, Fernandez-Sojo J, Genesca M, Falco V, Buzon MJ (2019) Latency reversal agents affect differently the latent reservoir present in distinct CD4+ T subpopulations. PLoS Pathog 15(8):e1007991. https://doi.org/10.1371/journal.ppat.1007991

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Cabrera C, Chang L, Stone M, Busch M, Wilson DH (2015) Rapid, fully automated digital immunoassay for p24 protein with the sensitivity of nucleic acid amplification for detecting acute HIV infection. Clin Chem 61(11):1372–1380. https://doi.org/10.1373/clinchem.2015.243287

    Article  CAS  PubMed  Google Scholar 

  10. Chang L, Song L, Fournier DR, Kan CW, Patel PP, Ferrell EP, Pink BA, Minnehan KA, Hanlon DW, Duffy DC, Wilson DH (2013) Simple diffusion-constrained immunoassay for p24 protein with the sensitivity of nucleic acid amplification for detecting acute HIV infection. J Virol Methods 188(1–2):153–160. https://doi.org/10.1016/j.jviromet.2012.08.017

    Article  CAS  PubMed  Google Scholar 

  11. Passaes C, Delagreverie HM, Avettand-Fenoel V, David A, Monceaux V, Essat A, Muller-Trutwin M, Duffy D, De Castro N, Wittkop L, Rouzioux C, Molina JM, Meyer L, Delaugerre C, Saez-Cirion A (2021) Ultrasensitive detection of p24 in plasma samples from people with primary and chronic HIV-1 infection. J Virol. https://doi.org/10.1128/JVI.00016-21

  12. Wu G, Cheney C, Huang Q, Hazuda DJ, Howell BJ, Zuck P (2021) Improved detection of HIV gag p24 protein using a combined immunoprecipitation and digital ELISA method. Front Microbiol 12:636703. https://doi.org/10.3389/fmicb.2021.636703

    Article  PubMed  PubMed Central  Google Scholar 

  13. Stuelke EL, James KS, Kirchherr JL, Allard B, Baker C, Kuruc JD, Gay CL, Margolis DM, Archin NM (2020) Measuring the inducible, replication-competent HIV reservoir using an ultra-sensitive p24 readout, the digital ELISA viral outgrowth assay. Front Immunol 11:1971. https://doi.org/10.3389/fimmu.2020.01971

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Bosque A, Nilson KA, Macedo AB, Spivak AM, Archin NM, Van Wagoner RM, Martins LJ, Novis CL, Szaniawski MA, Ireland CM, Margolis DM, Price DH, Planelles V (2017) Benzotriazoles reactivate latent HIV-1 through inactivation of STAT5 SUMOylation. Cell Rep 18(5):1324–1334. https://doi.org/10.1016/j.celrep.2017.01.022

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Macedo AB, Novis CL, De Assis CM, Sorensen ES, Moszczynski P, Huang SH, Ren Y, Spivak AM, Jones RB, Planelles V, Bosque A (2018) Dual TLR2 and TLR7 agonists as HIV latency-reversing agents. JCI Insight 3(19). https://doi.org/10.1172/jci.insight.122673

  16. Puertas MC, Bayon-Gil A, Garcia-Guerrero MC, Salgado M, Urrea V, Moron-Lopez S, Pena R, Jimenez-Moyano E, Clotet B, Prado JG, Martinez-Picado J (2021) Erratum for Puertas et al., “VIP-SPOT: an innovative assay to quantify the productive HIV-1 reservoir in the monitoring of cure strategies”. mBio 12(5):e0253521. https://doi.org/10.1128/mBio.02535-21

    Article  PubMed  Google Scholar 

  17. Lambert J, Mendes M, Yan Y-X, Ball A (2019) A universal planar assay format for high sensitivity cytokine quantification in human serum and plasma. J Immunol 202(1 Supplement):131.111

    Google Scholar 

  18. Cheng J, Myers TG, Levinger C, Kumar P, Kumar J, Goshu BA, Bosque A, Catalfamo M (2022) IL-27 induces IFN/STAT1-dependent genes and enhances function of TIGIT(+) HIVGag-specific T cells. iScience 25(1):103588. https://doi.org/10.1016/j.isci.2021.103588

    Article  CAS  PubMed  Google Scholar 

  19. Innis EA, Levinger C, Szaniawski MA, Williams E, Alcami J, Bosque A, Schiffer JT, Coiras M, Spivak AM, Planelles V (2021) Pharmacologic control of homeostatic and antigen-driven proliferation to target HIV-1 persistence. Biochem Pharmacol 194:114816. https://doi.org/10.1016/j.bcp.2021.114816

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Levinger C, Howard J, Cheng J, Tang P, Joshi A, Catalfamo M, Bosque A (2021) An ultrasensitive planar array p24 Gag ELISA to detect HIV-1 in diverse biological matrixes. Sci Rep 11(1):23682. https://doi.org/10.1038/s41598-021-03072-7

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgments

The work in A.B.’s lab is currently supported by the National Institute of Allergy and Infectious Diseases and National Institute of Health grants R01-AI154518, R01-AI147812, UM1-AI164565, T32-AI158105, R21-AI172042, R61-AI176582, and P30-AI117970

Author information

Authors and Affiliations

  1. Department of Microbiology, Immunology and Tropical Medicine, George Washington University, Washington, DC, USA

    Callie Levinger, J. Natalie Howard & Alberto Bosque

Authors
  1. Callie Levinger
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. J. Natalie Howard
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Alberto Bosque
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Alberto Bosque .

Editor information

Editors and Affiliations

  1. Albert Einstein College of Medicine, Bronx, NY, USA

    Vinayaka R. Prasad

  2. Albert Einstein College of Medicine, Bronx, NY, USA

    Ganjam V. Kalpana

Rights and permissions

Reprints and permissions

Copyright information

© 2024 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Levinger, C., Natalie Howard, J., Bosque, A. (2024). An Ultrasensitive p24 Assay to Measure HIV-1 in Diverse Biological Matrixes. In: Prasad, V.R., Kalpana, G.V. (eds) HIV Protocols . Methods in Molecular Biology, vol 2807. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3862-0_23

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-3862-0_23

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3861-3

  • Online ISBN: 978-1-0716-3862-0

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation