Liska V, Khimani A H, Hofmann-Lehmann R, Fink A N, Vlasak J, Ruprecht R M
Laboratory of Viral Pathogenesis, Dana-Farber Cancer Institute, Boston, Massachusetts 02115, USA.
AIDS Res Hum Retroviruses. 1999 Mar 20;15(5):445-50. doi: 10.1089/088922299311196.
We have succeeded in stably maintaining the entire genome of SIVmac239 as a plasmid clone. Supercoiled proviral plasmid DNA was inoculated intramuscularly into two adult rhesus macaques and into a neonate. All three animals became viremic and seroconverted. Viral kinetics were followed prospectively by quantitative competitive reverse transcriptase polymerase chain reaction (QC-RT-PCR), measurement of proviral DNA load in peripheral blood mononuclear cells (PBMCs) by PCR, and virus isolation by cocultivation. The infant developed high virus loads and succumbed to AIDS and SIV-associated nephropathy at 10 weeks postinoculation. Both adults are still living but have progressed to AIDS; one adult has also developed severe thrombocytopenia. We conclude that infection through intramuscular inoculation of cloned plasmid DNA encoding the entire proviral genome is reproducible and will provide a useful tool for studying viral pathogenesis.
我们已成功将SIVmac239的完整基因组作为质粒克隆稳定保存。将超螺旋前病毒质粒DNA肌肉注射到两只成年恒河猴和一只新生猴体内。所有三只动物都出现了病毒血症并发生了血清转化。通过定量竞争逆转录聚合酶链反应（QC-RT-PCR）、通过PCR测量外周血单个核细胞（PBMC）中的前病毒DNA载量以及通过共培养进行病毒分离，对病毒动力学进行了前瞻性跟踪。该婴儿在接种后10周出现高病毒载量，并死于艾滋病和与SIV相关的肾病。两只成年猴仍然存活，但已发展为艾滋病；其中一只成年猴还出现了严重的血小板减少症。我们得出结论，通过肌肉注射编码完整前病毒基因组的克隆质粒DNA进行感染是可重复的，这将为研究病毒发病机制提供一个有用的工具。
