HIV Protocols pp 281-292 | Cite as

Analysis of ABCA1 and Cholesterol Efflux in HIV-Infected Cells

  • Nigora Mukhamedova
  • Beda Brichacek
  • Christina Darwish
  • Anastas Popratiloff
  • Dmitri Sviridov
  • Michael BukrinskyEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1354)


Cholesterol is an essential component of the cellular membranes and, by extension, of the HIV envelope membrane, which is derived from the host cell plasma membrane. Depletion of the cellular cholesterol has an inhibitory effect on HIV assembly, reduces infectivity of the produced virions, and makes the cell less susceptible to HIV infection. It is not surprising that the virus has evolved to gain access to cellular proteins regulating cholesterol metabolism. One of the key mechanisms used by HIV to maintain high levels of cholesterol in infected cells is Nef-mediated inhibition of cholesterol efflux and the cholesterol transporter responsible for this process, ABCA1. In this chapter, we describe methods to investigate these effects of HIV-1 infection.

Key words

HIV-1 Nef Cholesterol efflux ABCA1 Confocal microscopy Image analysis 



This study was supported by NIH grants R01HL093818, R01HL101274, R21AI108533, P30AI087714, P30HD040677, S10OD010710, 1S10RR025565.


  1. 1.
    Waheed AA, Freed EO (2009) Lipids and membrane microdomains in HIV-1 replication. Virus Res 143:162–176CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Ono A, Freed EO (2001) Plasma membrane rafts play a critical role in HIV-1 assembly and release. Proc Natl Acad Sci U S A 98:13925–13930CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Manes S, del Real G, Lacalle RA et al (2000) Membrane raft microdomains mediate lateral assemblies required for HIV-1 infection. EMBO Rep 1:190–196CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Koseki M, Hirano K-i, Masuda D et al (2007) Increased lipid rafts and accelerated lipopolysaccharide-induced tumor necrosis factor-{alpha} secretion in Abca1-deficient macrophages. J Lipid Res 48:299–306CrossRefPubMedGoogle Scholar
  5. 5.
    Landry YD, Denis M, Nandi S et al (2006) ATP-binding cassette transporter A1 expression disrupts raft membrane microdomains through its ATPase-related functions. J Biol Chem 281:36091–36101CrossRefPubMedGoogle Scholar
  6. 6.
    Cui HL, Grant A, Mukhamedova N et al (2012) HIV-1 Nef mobilizes lipid rafts in macrophages through a pathway that competes with ABCA1-dependent cholesterol efflux. J Lipid Res 53:696–708CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Brichacek B, Darwish C, Popratiloff A et al (2014) HIV-1 Infection of macrophages induces retention of cholesterol transporter ABCA1 in the endoplasmic reticulum. AIDS Res Hum Retroviruses 30:947CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Jennelle L, Hunegnaw R, Dubrovsky L et al (2014) HIV-1 protein nef inhibits activity of ATP binding cassette transporter A1 by targeting endoplasmic reticulum chaperone calnexin. J Biol Chem 289:28870CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Asztalos BF, Mujawar Z, Morrow MP et al (2010) Circulating Nef induces dyslipidemia in simian immunodeficiency virus-infected macaques by suppressing cholesterol efflux. J Infect Dis 202:614–623CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Cui HL, Ditiatkovski M, Kesani R et al (2014) HIV protein Nef causes dyslipidemia and formation of foam cells in mouse models of atherosclerosis. FASEB J 28:2828–2839CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Crowe SM, Westhorpe CL, Mukhamedova N et al (2010) The macrophage: the intersection between HIV infection and atherosclerosis. J Leukoc Biol 87:589–598CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Neufeld EB, Remaley AT, Demosky SJ et al (2001) Cellular localization and trafficking of the human ABCA1 transporter. J Biol Chem 276:27584–27590CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Nigora Mukhamedova
    • 1
  • Beda Brichacek
    • 2
  • Christina Darwish
    • 2
  • Anastas Popratiloff
    • 3
  • Dmitri Sviridov
    • 1
  • Michael Bukrinsky
    • 2
    Email author
  1. 1.Baker IDI Heart and Diabetes InstituteMelbourneAustralia
  2. 2.Department of Microbiology, Immunology and Tropical MedicineGeorge Washington University School of Medicine and Health SciencesWashingtonUSA
  3. 3.Office of VP for ResearchGeorge Washington University Center for Microscopy and Image AnalysisWashingtonUSA

Personalised recommendations