Ex vivo analysis of SIV-infected cells by flow cytometry.
Deciphering the complex interactions between human and simian immunodeficiency viruses (HIV/SIV) and their host cells is crucial to the development of improved therapies and vaccines. Investigating these relationships has been complicated by the inability to directly analyze infected cells among freshly isolated peripheral blood lymphocytes. Here, we describe a method to detect cells productively infected with SIVmac239 ex vivo from the blood or lymph nodes by flow cytometry. Using this method, we show a close correlation between the frequency of productively infected cells in both sample type and the plasma viral load. We define that the minimum threshold for detecting productively infected cells in lymph nodes by flow cytometry requires a plasma virus concentration of ∼2.5 × 10(4) vRNA copy Equivalents (Eq)/ml. Conversely, an approximately 2 logs higher plasma viral load is needed to detect productively infected cells in the peripheral blood. This novel protocol provides a direct analytical tool to assess interactions between SIV and host cells, which is of key importance to investigators in AIDS research.
Reynolds
Matthew R
MR
AIDS Vaccine Research Laboratory, University of Wisconsin-Madison, Madison, Wisconsin 53711, USA.
Piaskowski
Shari M
SM
Weisgrau
Kimberly L
KL
Weiler
Andrea M
AM
Friedrich
Thomas C
TC
Rakasz
Eva G
EG
D000818:Animals; D002469:Cell Separation; D005434:Flow Cytometry / Q000379:methods*; D054884:Host-Pathogen Interactions / Q000502:physiology; D057230:Limit of Detection; D008198:Lymph Nodes / Q000473:pathology / Q000821:virology; D008214:Lymphocytes / Q000473:pathology / Q000821:virology; D008253:Macaca mulatta; D012367:RNA, Viral / Q000032:analysis; D016097:Simian Acquired Immunodeficiency Syndrome / Q000097:blood / Q000473:pathology*; D015302:Simian Immunodeficiency Virus / Q000235:genetics / Q000302:isolation & purification*; D019562:Viral Load
D012367:RNA, Viral