Prolonged adherence of human immunodeficiency virus-derived vector particles to hematopoietic target cells leads to secondary transduction in vitro and in vivo
Prolonged adherence of human immunodeficiency virus-derived vector particles to hematopoietic target cells leads to secondary transduction in vitro and in vivo
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DOI：
10.1128/jvi.01089-06
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发表时间：
2007-01-01
影响因子：
5.4
通讯作者：
Kurre, Peter
中科院分区：
文献类型：
--
作者：
Pan, Yung-Wei;Scarlett, Jarrad M.;Kurre, Peter
携带水泡性口炎病毒G（VSV-G）包膜糖蛋白的人类免疫缺陷病毒1型衍生慢病毒载体表现出广泛的宿主范围，并能稳定地覆盖静止的造血干细胞。鉴于对生物安全性和潜在生殖系传播的担忧，它们主要用于体外策略，被认为是确保去除过量的表面结合颗粒并防止体内传播。这里提出的研究，而不是揭示延长颗粒粘附后，离体暴露，尽管连续洗涤程序，与随后的转导的第二靶细胞在直接和transwell共培养。我们探讨了影响颗粒保留和转移的关键参数，并表明附着在细胞表面选择性地保护病毒颗粒免受血清补体介导的灭活。此外，对非骨髓清除的小鼠受体的研究表明，移植载体暴露的、洗涤的造血细胞导致功能性VSV-G/慢病毒载体颗粒的全身性传播。我们通过载体颗粒的无意转移和转基因产物在受体组织中的长期表达来证明遗传标记。我们的研究结果对生物安全、载体设计和细胞生物学研究具有重要意义。
Human immunodeficiency virus type 1-derived lentivirus vectors bearing the vesicular stomatitis virus G (VSV-G) envelope glycoprotein demonstrate a wide host range and can stably transduce quiescent hematopoietic stem cells. In light of concerns about biosafety and potential germ line transmission, they have been used predominantly for ex vivo strategies, thought to ensure the removal of excess surface-bound particles and prevent in vivo dissemination. Studies presented here instead reveal prolonged particle adherence after ex vivo exposure, despite serial wash procedures, with subsequent transduction of secondary target cells in direct and transwell cocultures. We explored the critical parameters affecting particle retention and transfer and show that attachment to the cell surface selectively protects virus particles from serum complement-mediated inactivation. Moreover, studies with nonmyeloablated murine recipients show that transplantation of vector-exposed, washed hematopoietic cells results in systemic dissemination of functional VSV-G/lentivector particles. We demonstrate genetic marking by inadvertent transfer of vector particles and prolonged expression of transgene product in recipient tissues. Our findings have implications for biosafety, vector design, and cell biology research.