Skip to main content

Use of a Macrophage-Tropic GFP-Tagged Human Immunodeficiency Virus Type 1 (HIV-1) to Study Viral Reservoirs

  • Protocol
Viral Applications of Green Fluorescent Protein

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 515))

Summary

The human immunodeficiency virus type 1 (HIV-1) predominantly infects two main cell types: T-lymphocytes and monocyte-derived cells such as macrophages, dendritic cells, and Langerhans cells. Studies in patients aimed at uncovering and understanding the molecular mechanisms for viral persistence and latency in macrophages in particular are challenging because these cells reside in tissues and can only be extracted using invasive methods. The in vitro human monocyte-derived macrophage (MDM) model provides an amenable system to study HIV–host cell interactions at the molecular level. Infection of macrophages with a recombinant M-tropic virus that contains the green fluorescent protein (GFP) gene within its genome allows the detection of cells with integrated viral DNA that are producing viral particles. The infected cells can be studied at the single-cell level using a variety of fluorescence-based technologies such as flow cytometry, conventional and confocal microscopy, and laser capture microdissection.

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

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 159.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Chun, T. W., R. T. J. Davey, D. Engel, H. C. Lane, and A. S. Fauci. (1999). Re-emergence of HIV after stopping therapy. Nature 40:874–875.

    Article  Google Scholar 

  2. Davey, R. T. J., N. Bhat, C. Yoder, T. W. Chun, J. A. Metcalf, R. Dewar, V. Natarajan, R. A. Lempicki, J. W. Adelsberger, K. D. Miller, J. A. Kovacs, M. A. Polis, R. E. Walker, J. Falloon, H. Masur, D. Gee, M. Baseler, D. S. Dimitrov, A. S. Fauci, and H. C. Lane. (1999). HIV-1 and T cell dynamics after interruption of highly active antiretroviral therapy (HAART) in patients with a history of sustained viral suppression. Proc. Natl. Acad. Sci. USA 96:15109–15114.

    Article  PubMed  CAS  Google Scholar 

  3. Harrigan, P. R., M. Whaley, and J. S. G. Montaner. (1999). Rate of HIV-1 RNA rebound upon stopping antiretroviral therapy. AIDS 13:F59–F62.

    Article  PubMed  CAS  Google Scholar 

  4. Brown, A., H. Zhang, P. Lopez, C. P. Pardo, and S. Gartner. (2006). In vitro modeling of the HIV-macrophage reservoir. J. Leukoc. Biol. 80:1127–1135.

    Article  PubMed  CAS  Google Scholar 

  5. Honda, Y., L. Rogers, K. Nakata, B. Y. Zhao, R. Pine, Y. Nakai, K. Kurosu, W. N. Rom, and M. Weiden. (1998). Type I interferon induces inhibitory 16-kD CCAAT/enhancer binding protein (C/EBP)beta, repressing the HIV-1 long terminal repeat in macrophages: pulmonary tuberculosis alters C/EBP expression, enhancing HIV-1 replication. J. Exp. Med. 188:1255–1265.

    Article  PubMed  CAS  Google Scholar 

  6. Hoshino, Y., K. Nakata, S. Hoshino, Y. Honda, D. B. Tse, T. Shioda, W. N. Rom, and M. Weiden. (2002). Maximal HIV-1 replication in alveolar macrophages during tuberculosis requires both lympho-cytes contact and cytokines. J. Exp. Med. 195:495–505.

    Article  PubMed  CAS  Google Scholar 

  7. Brown, A., S. Gartner, T. Kawano, N. Benoit, and C. Cheng-Mayer. (2005). HLA-A2 down-regulation on primary human macrophages infected with an M-tropic EGFP-tagged HIV-1 reporter virus. J. Leukoc. Biol. 78:675–685.

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

The author thanks Peter Lopez and Hao Zhang for their expertise in flow sorting, Carlos A. Pardo for use of the PALM Microlaser, and Suzanne Gartner for sharing her love and knowledge of monocyte–macrophage culturing and for making these studies possible. This work was supported by grants from The Campbell Foundation (A. Brown), National Institutes of Health R01 AI057007 (S. Gartner), and K08 DA16160 (C. Pardo).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Amanda M. Brown .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2009 Humana Press, a part of Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Brown, A.M. (2009). Use of a Macrophage-Tropic GFP-Tagged Human Immunodeficiency Virus Type 1 (HIV-1) to Study Viral Reservoirs. In: Hicks, B.W. (eds) Viral Applications of Green Fluorescent Protein. Methods in Molecular Biology™, vol 515. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-59745-559-6_11

Download citation

  • DOI: https://doi.org/10.1007/978-1-59745-559-6_11

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-934115-87-9

  • Online ISBN: 978-1-59745-559-6

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics