Isolation and HIV-1 infection of primary human microglia from fetal and adult tissue.

Glial inflammation, principally involving astrocytes and microglia, underlies the pathogenesis of a broad range of neurodegenerative disorders, including, most notably, human immunodeficiency virus (HIV-1)-associated dementia. Indeed, for the latter, disease mechanisms are attributed to viral infection and activation of microglia and perivascular macrophages and their resultant neurotoxic activities. Although monocyte-derived macrophages have served as models for microglia, they are limited both qualitatively and quantitatively in their immune responses and susceptibility to viral infection. Thus, the acquisition of primary human microglial cells is critical for laboratory studies of human neurological disease. In this chapter, we provide detailed methods of isolation, cultivation, characterization, HIV-1 infection, and experimental applications of primary human fetal and adult microglial cells, with particular emphasis on studies of HIV-1 neuropathogenesis.