Monocytes play an important, yet only partly understood, role in HIV-1 pathogenesis. Two main subsets of peripheral blood monocytes have been described; the major subset of monocytes are phenotypically characterized as being CD14hi/CD16a??, and a minor subset (5a??15 of total monocytes in healthy individuals), which are CD14lo/CD16hi, have been reported to be expanded in HIV-infected individuals. These CD14lo/CD16hi monocytes differ from the majority of monocytes in a number of ways, including the molecules expressed on their surface and how they function. Here we describe a flow-cytometric assay to identify and compare the expression of a representative surface molecule (CCR5) on CD14hi/CD16a?? and CD14lo/CD16hi monocytes in small volumes of whole blood, and methods to isolate monocyte subsets by both fluorescence-activated cell sorting (FACS) and magnetic bead sorting.
|Title of host publication||Methods in Molecular Biology - Human Retrovirus Protocols: Virology and Molecular Biology|
|Place of Publication||USA|
|Pages||343 - 353|
|Number of pages||11|
|Publication status||Published - 2005|