Inactivation of HIV-infected H9 cells in whole blood preparations by lysing/fixing reagents used in flow cytometry.
Reagents that lyse red blood cells and fix white blood cells were tested for their ability to inactivate cell-associated human immunodeficiency virus (HIV). Whole blood was spiked with cells from an HIV-positive cell line (H9), lysed, and fixed. The cell preparations were then cocultured with T cell blasts in serial ten-fold dilutions to rescue infectious virus and measure viral titer. All commercial lysing and fixing reagents tested inactivated cell-associated HIV by 3-5 logs, while ammonium chloride had little effect. Although an additional incubation with 1% formaldehyde for 30 min did not increase the effectiveness of the commercial lysing/fixing reagents, it did inactivate cell-associated HIV in blood treated with ammonium chloride.
Nicholson
J K
JK
Immunology Branch, Centers for Disease Control, Atlanta, GA 30333.
Browning
S W
SW
Orloff
S L
SL
McDougal
J S
JS
D016923:Cell Death; D002478:Cells, Cultured; D005404:Fixatives; D005434:Flow Cytometry / Q000379:methods*; D005557:Formaldehyde / Q000737:chemistry; D006678:HIV / Q000187:drug effects*; D015658:HIV Infections / Q000382:microbiology*; D006801:Humans; D016130:Immunophenotyping / Q000379:methods*; D066298:In Vitro Techniques
D005404:Fixatives; D005557:Formaldehyde