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Ex vivo analysis of SIV-infected cells by flow cytometry

✍ Scribed by Matthew R. Reynolds; Shari M. Piaskowski; Kimberly L Weisgrau; Andrea M. Weiler; Thomas C. Friedrich; Eva G. Rakasz


Book ID
102815163
Publisher
John Wiley and Sons
Year
2010
Tongue
English
Weight
513 KB
Volume
77A
Category
Article
ISSN
0196-4763

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✦ Synopsis


Abstract

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 Γ— 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. Β© 2010 International Society for Advancement of Cytometry.


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