Chambers R F, Terada M, Kufe D, Ohno T
Nissin Molecular Biology Institute, Boston, MA.
AIDS Res Hum Retroviruses. 1991 May;7(5):459-63. doi: 10.1089/aid.1991.7.459.
The shaking method of harvesting human immunodeficiency virus type 1 (HIV-1) is a powerful method of obtaining high titer, highly infective virus solutions. In this method infected cells are suspended in a small volume of liquid and the mixture is shaken. Viral infectivity, measured by tissue culture infective dose (TCID50) studies, rises faster than virus titer, as measured by reverse transcriptase levels. It is postulated that this disproportionate increase in infectivity results from improved infectivity for the virus particles obtained from shaking the infected cells. Of the five strains of HIV-1 studied (IIIB, AL1212, 906, RJ4029, and MN), one strain, MN, behaved differently than the others. Upon shaking, its virus titer increased 18-fold, as opposed to the 5-10 fold increase demonstrated by the other strains. These results may indicate that MN virions are retained more on the surface of the infected cells, rather than budding off into the surrounding medium, than other HIV-1 strains. In support of this theory it was found that ratios of immunofluorescence assay scores to reverse transcriptase levels were higher for MN than for other strains.
收获1型人类免疫缺陷病毒（HIV-1）的振荡法是一种获取高滴度、高感染性病毒溶液的有效方法。在该方法中，将感染的细胞悬浮于少量液体中并振荡混合物。通过组织培养感染剂量（TCID50）研究测定的病毒感染性比通过逆转录酶水平测定的病毒滴度上升得更快。据推测，这种感染性的不成比例增加是由于振荡感染细胞获得的病毒颗粒的感染性提高所致。在所研究的5株HIV-1（IIIB、AL1212、906、RJ4029和MN）中，有一株MN的表现与其他菌株不同。振荡后，其病毒滴度增加了18倍，而其他菌株仅增加了5至10倍。这些结果可能表明，与其他HIV-1菌株相比，MN病毒粒子更多地保留在感染细胞表面，而不是芽生到周围培养基中。支持这一理论的是，发现MN的免疫荧光测定分数与逆转录酶水平的比率高于其他菌株。
