Bixby Jacqueline G, Laur Olga, Johnson Welkin E, Desrosiers Ronald C
New England Primate Research Center, Harvard Medical School, Southborough, Massachusetts 01772-9102, USA.
AIDS Res Hum Retroviruses. 2010 Oct;26(10):1115-31. doi: 10.1089/aid.2010.0029. Epub 2010 Sep 13.
AIDS vaccine and pathogenesis research will benefit from a more diverse array of cloned SIV challenge stocks from which to choose. Toward this end, 20 envelope genes were cloned from an extensively used, primary stock of uncloned SIVmac251. Each of the 20 clones had a unique sequence. Their translated sequences differed by as many as 26 amino acids from one another and by as many as 45 amino acids from the commonly used clone SIVmac239. Envelope sequences up to and including the membrane-spanning domain were exchanged into the infectious pathogenic SIVmac239 clone and virus stocks were produced by HEK293T cell transfection. Seventeen of the 20 recombinants were replication competent. The infectivities per ng p27 of the 17 new replication-competent recombinants in C8166-SEAP cells and in TZM-bl cells ranged from minus 32-fold to plus 7.6-fold relative to SIVmac239. A range of sensitivities to neutralization by sCD4 and by sera from SIV-infected macaques was observed but none was as sensitive to these neutralizing agents as SIVmac316, the highly macrophage-competent derivative of SIVmac239. Four strains that were most sensitive to sCD4 inhibition were also among the most sensitive to antibody-mediated neutralization. None of the new recombinant viruses replicated as well as SIVmac316 in primary alveolar macrophage cultures from rhesus monkeys but three of the strains did exhibit significant levels of delayed replication in these primary macrophages, reaching peak levels of virus production of ≥50 ng/ml p27 compared to 600-800 ng/ml p27 with SIVmac316. These new SIV clones are being contributed to the NIH AIDS Reagent Repository and are available to the scientific community.
艾滋病疫苗和发病机制研究将受益于更多样化的克隆猴免疫缺陷病毒（SIV）攻击毒株以供选择。为此，从广泛使用的未克隆SIVmac251原始毒株中克隆了20个包膜基因。这20个克隆中的每一个都有独特的序列。它们的翻译序列彼此之间相差多达26个氨基酸，与常用克隆SIVmac239相差多达45个氨基酸。将直至并包括跨膜结构域的包膜序列交换到具有感染性的致病性SIVmac239克隆中，并通过人胚肾293T细胞转染产生病毒毒株。20个重组体中有17个具有复制能力。相对于SIVmac239，17个新的具有复制能力的重组体在C8166-SEAP细胞和TZM-bl细胞中每纳克p27的感染性范围为低32倍至高7.6倍。观察到对可溶性CD4（sCD4）和来自感染SIV的猕猴血清中和的一系列敏感性，但没有一个像SIVmac239的高度巨噬细胞适应性衍生物SIVmac316那样对这些中和剂敏感。对sCD4抑制最敏感的四个毒株也是对抗体介导的中和最敏感的毒株。在恒河猴的原代肺泡巨噬细胞培养物中，没有一种新的重组病毒复制得像SIVmac316那样好，但其中三个毒株在这些原代巨噬细胞中确实表现出显著水平的延迟复制，与SIVmac316的病毒产生峰值水平600 - 800纳克/毫升p27相比，达到≥50纳克/毫升p27的峰值水平。这些新的SIV克隆已被提供给美国国立卫生研究院（NIH）艾滋病试剂库，并可供科学界使用。
