Isolation and characterization of human cells resistant to retrovirus infection.
Isolation and characterization of human cells resistant to retrovirus infection.
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DOI：
10.1186/1742-4690-4-45
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发表时间：
2007-07-03
期刊：
影响因子：
3.3
通讯作者：
Somia NV
中科院分区：
文献类型：
--
作者：
Lech P;Somia NV
识别HIV-1感染所需的宿主细胞蛋白将增加我们对HIV-1生命周期的了解，并有助于开发抗击病毒感染的治疗方法。我们和其他研究人员已经对啮齿动物细胞进行了突变，并分离出了抗逆转录病毒感染的突变细胞系。由于VSVG假型HIV-1单轮感染对不同物种细胞的感染效率存在差异，我们进行了遗传筛选以分离出对HIV-1感染具有抵抗力的人细胞。我们对人类HeLa细胞进行了化学诱变，并验证了我们在测试二倍体基因座分离突变株的能力。然后，我们进行了一次筛选，以分离耐受编码有毒基因产物的HIV-1载体感染的HeLa细胞突变体。我们分离了两个突变的细胞系，它们对HIV-1载体感染的抵抗力高达10倍。我们已经证实，这些细胞对感染具有抵抗力，并且在基因表达方面没有缺陷。我们已经证实，抗性表型不是由于进入缺陷造成的。突变体和野生型细胞的融合实验证明，这两个克隆中产生抗性的突变是隐性的。我们还确定了这两个突变体的阻断性质。其中一个克隆在病毒RNA逆转录时或之前被阻断，第二个克隆的病毒DNA合成动力学受阻，前整合复合体的核输入受阻。可以分离出对HIV-1感染具有抵抗力的人类细胞突变株。这些突变体在遗传上是隐性的，并确定了宿主细胞因子可以靶向阻断HIV-1感染的两个点。
Identification of host cell proteins required for HIV-1 infection will add to our knowledge of the life cycle of HIV-1 and in the development of therapeutics to combat viral infection. We and other investigators have mutagenized rodent cells and isolated mutant cell lines resistant to retrovirus infection. Since there are differences in the efficiency of single round infection with VSVG pseudotyped HIV-1 on cells of different species, we conducted a genetic screen to isolate human cells resistant to HIV-1 infection. We chemically mutagenized human HeLa cells and validated our ability to isolate mutants at test diploid loci. We then executed a screen to isolate HeLa cell mutants resistant to infection by an HIV-1 vector coding for a toxic gene product. We isolated two mutant cell lines that exhibit up to 10-fold resistance to infection by HIV-1 vectors. We have verified that the cells are resistant to infection and not defective in gene expression. We have confirmed that the resistance phenotype is not due to an entry defect. Fusion experiments between mutant and wild-type cells have established that the mutations conferring resistance in the two clones are recessive. We have also determined the nature of the block in the two mutants. One clone exhibits a block at or before reverse transcription of viral RNA and the second clone has a retarded kinetic of viral DNA synthesis and a block at nuclear import of the preintegration complex. Human cell mutants can be isolated that are resistant to infection by HIV-1. The mutants are genetically recessive and identify two points where host cell factors can be targeted to block HIV-1 infection.
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DOI：
10.1038/nrd1853
发表时间：
2005-10
期刊：
Nature reviews. Drug discovery
影响因子：
--
作者：
Kwong AD;Rao BG;Jeang KT
通讯作者：
Jeang KT
影响因子：
5.4
作者：
Camaur, D;Trono, D
通讯作者：
Trono, D
影响因子：
5.4
作者：
Delelis, O;Saïb, A;Sonigo, P
通讯作者：
Sonigo, P
影响因子：
56.9
作者：
Naldini, L;Blomer, U;Trono, D
通讯作者：
Trono, D
影响因子：
3.3
作者：
Gao, GX;Goff, SP
通讯作者：
Goff, SP