Cameron P U, Pagnon J C, van Baare J, Reece J C, Vardaxis N J, Crowe S M
Department of Microbiology and Immunology, University of Melbourne, Victoria, Australia.
J Med Virol. 2000 Feb;60(2):182-8. doi: 10.1002/(sici)1096-9071(200002)60:2<182::aid-jmv13>3.0.co;2-y.
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 degrees 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 degrees C. HIV-1 was recovered consistently from skin stored in 85% glycerol at 4 degrees C for up to 72 hr but virus isolation was infrequent after storage for more than 5 days. At 20 degrees C or 37 degrees 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 degrees C eliminated effectively HIV-1 from skin.
同种异体断层皮片移植在烧伤治疗中被广泛应用。皮肤甘油保存法相对简便，这表明它将在HIV-1血清阳性率高的地区得到越来越多的应用。使用嗜巨噬细胞株HIV-1作为无细胞病毒悬液、在感染的外周血单个核细胞（PBMC）内或在体外HIV-1感染的新鲜尸体断层皮片内，测定甘油保存法使体外感染的皮肤移植片中存在的HIV-1失活的能力。采用不同的温度和甘油浓度，并通过与丝裂原激活的外周血单个核细胞（PBMC）共培养以及7-10天后测定逆转录酶活性来确定感染性。无细胞HIV-1在4℃下于70%或更高浓度的甘油中30分钟内失活。在类似暴露条件下，细胞或皮肤相关的HIV-1滴度降低，但在4℃下用70%和85%的甘油处理时并未消除。在4℃下于85%甘油中保存长达72小时的皮肤中可始终回收HIV-1，但保存超过5天后病毒分离很少见。在20℃或37℃下，70%或85%的甘油可在8小时内使细胞或皮肤相关的HIV-1失活。最初的甘油处理程序和在4℃下保存有效地从皮肤中消除了HIV-1。
