Efficacy and Kinetics of Glycerol Inactivation of HIV-1 in Split Skin Grafts

Allogeneic split skin grafts are used widely in the treatment of burns. The relative simplicity of glycerol preservation of skin suggests it will be used increasingly in areas of high HIV-1 seroprevalence. The ability of glycerol preservation to inactivate HIV-1 present in skin graft infected in vitro was determined using a macrophage tropic strain HIV-1 as a cell-free virus suspension, within infected PBMCs, or within in vitro HIV-1 infected fresh cadaveric split skin. Different temperatures and concentrations of glycerol were used and infectivity determined by coculture with mitogen activated peripheral blood mononuclear cells (PBMCs) and measurement of reverse transcriptase activity after 7-10 days. Cell-free HIV-1 was inactivated within 30 min at 4 degree C in glycerol concentrations of 70% or higher. During similar exposure cell- or skin-associated HIV-1 titer was reduced but not eliminated with 70% and 85% glycerol at 4 degree C. HIV-1 was recovered consistently from skin stored in 85% glycerol at 4 degree C for up to 72 hr but virus isolation was infrequent after storage for more than 5 days. At 20 degree C or 37 degree C, 70% or 85% glycerol could inactivate cell- or skin-associated HIV-1 within 8 hr. The initial glycerolization procedures and the storage at 4 degree C eliminated effectively HIV-1 from skin.