Measles virus glycoprotein-based lentiviral targeting vectors that avoid neutralizing antibodies.
Measles virus glycoprotein-based lentiviral targeting vectors that avoid neutralizing antibodies.
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DOI：
10.1371/journal.pone.0046667
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Buchholz CJ
中科院分区：
文献类型：
--
作者：
Kneissl S;Abel T;Rasbach A;Brynza J;Schneider-Schaulies J;Buchholz CJ
慢病毒载体（LV）是一种有效的基因转移载体，经常应用于研究和最近的临床试验。将LV进入重新靶向到感兴趣的细胞类型是提高基因转移安全性和有效性的关键问题。最近，我们已经开发了一种针对LV的靶向方法，该方法通过将工程麻疹病毒（MV）糖蛋白、负责受体识别的血凝素（H）和融合蛋白整合到它们的包膜中。H蛋白显示出对靶受体特异性的单链抗体（scFv），并且通过其胞外域中的点突变被消融以识别MV受体CD 46和SLAM。人体全身给药的一个潜在障碍是由于疫苗接种或自然感染而预先存在的MV特异性免疫。我们比较了靶向载体和非靶向载体在α-MV抗体阳性人血浆存在下的转导，所述非靶向载体在其胞外域中具有未修饰的MV糖蛋白（MV-LV）。在1∶160的血浆稀释度下，MV-LV几乎被完全中和，而靶向载体显示出60%至90%的相对转导效率。此外，在1∶80的血浆稀释度下，必须应用至少4倍更高的MV-LV感染复数（MOI）以获得与靶向载体类似的转导效率。此外，当载体归一化至其p24值时，靶向载体显示出对人血浆中α-MV抗体的部分保护。此外，具有接近H的受体结合位点的推定表位的单克隆中和抗体K71不中和靶向载体，但中和MV-LV。观察到的中和逃逸可能是由于H胞外域中的点突变可能破坏了抗体结合位点。此外，scFv介导的经由靶受体的细胞进入可在干扰经由天然MV受体进入的α-MV抗体存在下进行。这些结果对于靶向载体在人体中的体内应用是有希望的。
Lentiviral vectors (LVs) are potent gene transfer vehicles frequently applied in research and recently also in clinical trials. Retargeting LV entry to cell types of interest is a key issue to improve gene transfer safety and efficacy. Recently, we have developed a targeting method for LVs by incorporating engineered measles virus (MV) glycoproteins, the hemagglutinin (H), responsible for receptor recognition, and the fusion protein into their envelope. The H protein displays a single-chain antibody (scFv) specific for the target receptor and is ablated for recognition of the MV receptors CD46 and SLAM by point mutations in its ectodomain. A potential hindrance to systemic administration in humans is pre-existing MV-specific immunity due to vaccination or natural infection. We compared transduction of targeting vectors and non-targeting vectors pseudotyped with MV glycoproteins unmodified in their ectodomains (MV-LV) in presence of α-MV antibody-positive human plasma. At plasma dilution 1∶160 MV-LV was almost completely neutralized, whereas targeting vectors showed relative transduction efficiencies from 60% to 90%. Furthermore, at plasma dilution 1∶80 an at least 4-times higher multiplicity of infection (MOI) of MV-LV had to be applied to obtain similar transduction efficiencies as with targeting vectors. Also when the vectors were normalized to their p24 values, targeting vectors showed partial protection against α-MV antibodies in human plasma. Furthermore, the monoclonal neutralizing antibody K71 with a putative epitope close to the receptor binding sites of H, did not neutralize the targeting vectors, but did neutralize MV-LV. The observed escape from neutralization may be due to the point mutations in the H ectodomain that might have destroyed antibody binding sites. Furthermore, scFv mediated cell entry via the target receptor may proceed in presence of α-MV antibodies interfering with entry via the natural MV receptors. These results are promising for in vivo applications of targeting vectors in humans.
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影响因子：
20.3
作者：
Frecha, Cecilia;Costa, Caroline;Verhoeyen, Els
通讯作者：
Verhoeyen, Els
影响因子：
48
作者：
Anliker, Brigitte;Abel, Tobias;Buchholz, Christian J.
通讯作者：
Buchholz, Christian J.
影响因子：
5.4
作者：
LIEBERT, UG;FLANAGAN, SG;RIMA, BK
通讯作者：
RIMA, BK
DOI：
10.1038/nrg2141
发表时间：
2007-08
期刊：
Nature reviews. Genetics
影响因子：
--
作者：
Waehler R;Russell SJ;Curiel DT
通讯作者：
Curiel DT
影响因子：
64.8
作者：
Muhlebach, Michael D.;Mateo, Mathieu;Sinn, Patrick L.;Pruefer, Steffen;Uhlig, Katharina M.;Leonard, Vincent H. J.;Navaratnarajah, Chanakha K.;Frenzke, Marie;Wong, Xiao X.;Sawatsky, Bevan;Ramachandran, Shyam;McCray, Paul B., Jr.;Cichutek, Klaus;von Messling, Veronika;Lopez, Marc;Cattaneo, Roberto
通讯作者：
Cattaneo, Roberto