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J Neurovirol. 2017 Feb;23(1):47-66. doi: 10.1007/s13365-016-0499-3. Epub 2016 Nov 21.

Immortalization of primary microglia: a new platform to study HIV regulation in the central nervous system.

Author information

1
Department of Molecular Biology and Microbiology, Case Western Reserve University, Cleveland, OH, 44106, USA.
2
Department of Neurosciences, Case Western Reserve University, Cleveland, OH, 44106, USA.
3
Department of Biochemistry, Case Western Reserve University, Cleveland, OH, 44106, USA.
4
Department of Molecular Biology and Microbiology, Case Western Reserve University, Cleveland, OH, 44106, USA. dxa150@case.edu.

Abstract

The major reservoirs for HIV in the CNS are in the microglia, perivascular macrophages, and to a lesser extent, astrocytes. To study the molecular events controlling HIV expression in the microglia, we developed a reliable and robust method to immortalize microglial cells from primary glia from fresh CNS tissues and commercially available frozen glial cells. Primary human cells, including cells obtained from adult brain tissue, were transformed with lentiviral vectors expressing SV40 T antigen or a combination of SVR40 T antigen and hTERT. The immortalized cells have microglia-like morphology and express key microglial surface markers including CD11b, TGFβR, and P2RY12. Importantly, these cells were confirmed to be of human origin by sequencing. The RNA expression profiles identified by RNA-seq are also characteristic of microglial cells. Furthermore, the cells demonstrate the expected migratory and phagocytic activity, and the capacity to mount an inflammatory response characteristic of primary microglia. The immortalization method has also been successfully applied to a wide range of microglia from other species (macaque, rat, and mouse). To investigate different aspects of HIV molecular regulation in CNS, the cells have been superinfected with HIV reporter viruses and latently infected clones have been selected that reactive HIV in response to inflammatory signals. The cell lines we have developed and rigorously characterized will provide an invaluable resource for the study of HIV infection in microglial cells as well as studies of microglial cell function.

KEYWORDS:

HIV regulation; HIV-associated neurocognitive disorders; Microglia; NeuroAIDS

PMID:
27873219
PMCID:
PMC5329090
DOI:
10.1007/s13365-016-0499-3
[Indexed for MEDLINE]
Free PMC Article

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