Published: Vol 9, Iss 13, Jul 5, 2019 DOI: 10.21769/BioProtoc.3289 Views: 5209
Reviewed by: Lokesh KalekarPooja SaxenaAnonymous reviewer(s)
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Abstract
Monocyte infection by HIV-1 is an important component of chronic HIV pathogenesis. Following infection by HIV-1, monocytes are able to cross the blood brain barrier and set up a viral reservoir in the central nervous system. Additionally, in the setting of chronic HIV-1 infection, monocytes can become activated either directly through HIV-1 infection or indirectly via HIV-1-mediated systemic immune activation. Currently, there are few studies looking at HIV-1 infection of primary human monocytes in vitro. Furthermore, detection of successful HIV-1 infection of monocytes can be laborious requiring an ELISA for p24 or assessing levels of HIV-1 mRNA or DNA. This protocol utilizes an HIV-1 strain expressing GFP to allow for easy quantification of HIV-1 infection by fluorescence-assisted cell sorting (FACS). By determining HIV-1 infection by FACS one can take advantage of its multiparametric nature allowing for the use of less cells and the ability to assess the expression of other markers on HIV-1+ and HIV-1- cells in the same experiment. Additionally, this protocol could be modified to study HIV-1 infection of other cells including CD4+ T cells.
Keywords: MonocytesBackground
This protocol will be of use to those looking to perform studies assessing the role that myeloid cells play in HIV-1 infection. Myeloid cells, in particular, monocytes, have been recognized as crucial mediators of chronic and acute inflammation in HIV-1 infection (Burdo et al., 2013; Anzinger et al., 2014; Merino et al., 2017). Gaining a deeper understanding of the interactions between the HIV-1 virus and monocytes will be fundamental in advancing our knowledge on how HIV-1 infection produces a chronic state of immune activation, even in the presence of combined anti-retroviral therapy (Anzinger et al., 2014). With persons infected by HIV-1 now exhibiting lower death rates, we are just now seeing the long-term effects of HIV-1-induced chronic immune activation which can present as: HIV-associated neurocognitive disorder (HAND), hepatic steatosis, renal failure, hepatitis, atherosclerosis, insulin resistance, osteoporosis, and more (Cardoso et al., 2013; Katlama et al., 2013).
The method presented here will be useful to both study how HIV-1 infection of monocytes alters cellular function, and to facilitate the development of novel methods to modulate or prevent HIV-1 infection of monocytes. In particular, this protocol is advantageous in that it allows for quick and easy analysis of monocyte HIV-1 infection using FACS without the need to intracellular stain for p24.
Materials and Reagents
Equipment
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Acknowledgments
This work was supported by the NIH in the form of an R21 grant to YAA (1R21AI122808-01). We also thank Sarah Godbehere for her critical review of this manuscript. This method is adapted from Wang et al., 2018.
Competing interests
PO and YAA have a patent pending regarding modulation of SLAMF7 signaling for the treatment of inflammation. The authors declare no additional conflicts.
References
Article Information
Copyright
© 2019 The Authors; exclusive licensee Bio-protocol LLC.
How to cite
O’Connell, P., Zheng, Y., Amalfitano, A. and Aldhamen, Y. A. (2019). In vitro Infection of Primary Human Monocytes with HIV-1. Bio-protocol 9(13): e3289. DOI: 10.21769/BioProtoc.3289.
Category
Immunology > Immune cell staining > Flow cytometry
Microbiology > Microbe-host interactions > Virus
Cell Biology > Cell isolation and culture > Cell isolation
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