PEGylation of a vesicular stomatitis virus G pseudotyped lentivirus vector prevents inactivation in serum
PEGylation of a vesicular stomatitis virus G pseudotyped lentivirus vector prevents inactivation in serum
复制标题
DOI：
10.1128/jvi.78.2.912-921.2004
复制
发表时间：
2004-01-01
影响因子：
5.4
通讯作者：
Kobinger, GP
中科院分区：
文献类型：
--
作者：
Croyle, MA;Callahan, SM;Kobinger, GP
用于临床应用的水泡性口炎病毒G（VSV-G）假型慢病毒载体的一个缺点是载体被人血清补体灭活。为了防止这种情况，将单甲氧基聚（乙）二醇缀合至表达大肠杆菌β-半乳糖苷酶的VSV-G-人免疫缺陷病毒载体。该修饰不影响体外转导效率，并保护载体在补体活性的人和小鼠血清中免于失活。通过有限稀释测定法测定静脉内给予未修饰或PEG化病毒颗粒的小鼠的血液中的活性载体以评价转导效率，并测定p24（存在的病毒颗粒总数的指示物）。PEG化将活性载体的循环半衰期延长了5倍，并将血清中载体失活速率降低了1，000倍。循环中存在的病毒颗粒总数的药代动力学特征不受PEG化的影响。用聚（乙）二醇修饰载体显著增强了病毒全身给药后14天在骨髓和脾脏中的转导效率。这些结果与药代动力学曲线一致，表明PEG化确实保护病毒免于在血清中失活，因此，提高了VSV-G假型慢病毒载体在体内易感器官中的转导效率。
One disadvantage of vesicular stomatitis virus G (VSV-G) pseudotyped lentivirus vectors for clinical application is inactivation of the vector by human serum complement. To prevent this, monomethoxypoly(ethylene) glycol was conjugated to a VSV-G-human immunodeficiency virus vector expressing Escherichia coli beta-galactosidase. The modification did not affect transduction efficiency in vitro and protected the vector from inactivation in complement-active human and mouse sera. Blood from mice dosed intravenously with either the unmodified or the PEGylated virus particles was assayed for active vector by a limiting-dilution assay to evaluate transduction efficiency and for p24, an indicator of the total number of virus particles present. PEGylation extended the circulation half-life of active vector by a factor of 5 and reduced the rate of vector inactivation in the serum by a factor of 1,000. Pharmacokinetic profiles for the total number of virus particles present in the circulation were unaffected by PEGylation. Modification of the vector with poly(ethylene) glycol significantly enhanced transduction efficiency in the bone marrow and in the spleen 14 days after systemic administration of the virus. These results, in concert with the pharmacokinetic profiles, indicate that PEGylation does protect the virus from inactivation in the serum and, as a result, improves the transduction efficiency of VSV-G pseudotyped lentivirus vectors in susceptible organs in vivo.