HIV sequence variation associated with env antisense adoptive T-cell therapy in the hNSG mouse model.
HIV sequence variation associated with env antisense adoptive T-cell therapy in the hNSG mouse model.
复制标题
hNSG 小鼠模型中与 env 反义过继性 T 细胞治疗相关的 HIV 序列变异。
DOI：
10.1038/mt.2009.316
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发表时间：
2010
期刊：
影响因子：
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通讯作者：
Bushman,FredericD
中科院分区：
文献类型：
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作者：
Mukherjee,Rithun;Plesa,Gabriela;Sherrill-Mix,Scott;Richardson,MaxW;Riley,JamesL;Bushman,FredericD
慢病毒载体在人类中的首次使用涉及用人类免疫缺陷病毒（HIV）衍生的反义（envAS）载体转导成熟T细胞以保护细胞免受HIV感染。在这项研究中，只有少数患者的T细胞群体可以被基因修饰，这就提出了一个问题，即改变的细胞是否会影响复制的艾滋病毒群体。我们使用用~4- 11% envAS修饰的人T细胞重建的人源化NOD/SCID IL-2 R γnull（hNSG）小鼠对此进行了研究。用HIV-1 NL 4 -3或HIV-1BaL攻击小鼠，前者与envAS具有完全互补的envAS，后者与envAS具有完全互补的envAS。在接受denvAS修饰细胞的小鼠和未接受的对照小鼠之间，病毒滴度没有差异。使用454/Roche焦磷酸测序，我们分析了血清中HIV群体的突变谱-从33只小鼠中，我们回收了84，074个总读数，包括31，290个独特的序列变体。我们发现envAS处理的小鼠中A-G转换和缺失富集，这与之前的组织培养研究相似，其中大多数靶细胞含有envAS，尽管这里少数细胞是envAS修饰的。出乎意料的是，这种富集仅在用HIV-1BaL攻击后才被检测到，其中病毒基因组将与envAS形成不完美的双链体，而不是HIV-1 NL 4 -3，其中将形成完全匹配的双链体。
The first use of lentiviral vectors in humans involved transduction of mature T-cells with an human immunodeficiency virus (HIV)–derivedenvantisense (envAS) vector to protect cells from HIV infection. In that study, only a minority of the patient T-cell population could be gene-modified, raising the question of whether the altered cells could affect replicating HIV populations. We investigated this using humanized NOD/SCID IL-2Rγnull(hNSG) mice reconstituted with ~4–11%envAS-modified human T-cells. Mice were challenged with HIV-1NL4-3, which has anenvperfectly complementary toenvAS, or with HIV-1BaL, which has a divergentenv. No differences were seen in viral titer between mice that receivedenvAS-modified cells and control mice that did not. Using 454/Roche pyrosequencing, we analyzed the mutational spectrum in HIV populations in serum—from 33 mice we recovered 84,074 total reads comprising 31,290 unique sequence variants. We found enrichment of A-to-G transitions and deletions inenvAS-treated mice, paralleling a previous tissue culture study where most target cells containedenvAS, even though minority of cells wereenvAS-modified here. Unexpectedly, this enrichment was only detected after the challenge with HIV-1BaL, where the viral genome would form an imperfect duplex withenvAS, and not HIV-1NL4-3, where a perfectly matched duplex would form.