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Methods Mol Biol. 2016;1354:281-92. doi: 10.1007/978-1-4939-3046-3_19.

Analysis of ABCA1 and Cholesterol Efflux in HIV-Infected Cells.

Author information

1
Baker IDI Heart and Diabetes Institute, Melbourne, VIC, 3004, Australia.
2
Department of Microbiology, Immunology and Tropical Medicine, George Washington University School of Medicine and Health Sciences, Ross Hall, Room 624, 2300 Eye Street, N.W., Washington, DC, 20037, USA.
3
Office of VP for Research, George Washington University Center for Microscopy and Image Analysis, Washington, DC, 20037, USA.
4
Department of Microbiology, Immunology and Tropical Medicine, George Washington University School of Medicine and Health Sciences, Ross Hall, Room 624, 2300 Eye Street, N.W., Washington, DC, 20037, USA. mbukrins@gwu.edu.

Abstract

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.

KEYWORDS:

ABCA1; Cholesterol efflux; Confocal microscopy; HIV-1; Image analysis; Nef

PMID:
26714719
PMCID:
PMC5047010
DOI:
10.1007/978-1-4939-3046-3_19
[Indexed for MEDLINE]
Free PMC Article

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