Retrovirus-associated heparan sulfate mediates immobilization and gene transfer on recombinant fibronectin
Retrovirus-associated heparan sulfate mediates immobilization and gene transfer on recombinant fibronectin
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DOI：
10.1128/jvi.76.17.8722-8728.2002
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发表时间：
2002-09-01
影响因子：
5.4
通讯作者：
Andreadis, ST
中科院分区：
文献类型：
--
作者：
Lei, P;Bajaj, B;Andreadis, ST
重组逆转录病毒已被证明与纤连蛋白（FN）结合，并增加基因转移到各种细胞类型的效率。尽管最近的工作，以优化重组FN的基因转移，逆转录病毒结合FN和靶细胞与结合的病毒的相互作用的机制仍然难以捉摸。我们研究了病毒表面糖蛋白（gp 70），细胞条件培养基和蛋白多糖介导逆转录病毒结合FN的作用。我们还研究了聚凝胺（PB）在这些相互作用中的作用。我们发现gp 70不参与逆转录病毒与FN的结合。然而，病毒的固定化并不能克服其对受体的需求，成功的基因转移仍然需要gp 70。我们的研究结果清楚地表明，逆转录病毒结合FN通过病毒相关的硫酸乙酰肝素（HS）和结合是必要的转导没有PB。两种不同的基因转移模式取决于PB：（i）在PB存在下，逆转录病毒直接与靶细胞相互作用;（ii）在PB不存在下，逆转录病毒与FN结合，靶细胞与固定化病毒相互作用。PB可能通过与病毒HS相互作用，降低病毒颗粒的负电荷，从而促进前者的发生。有趣的是，后一种模式更有效，导致显著增强的基因转移。更好地了解这些相互作用可能会提供深入了解病毒-细胞相互作用，并导致更合理的设计转导方案。
Recombinant retroviruses have been shown to bind to fibronectin (FN) and increase the efficiency of gene transfer to a variety of cell types. Despite recent work to optimize gene transfer on recombinant FN, the mechanism of retrovirus binding to FN and the interactions of target cells with the bound virus remain elusive. We investigated the roles of virus surface glycoprotein (gp70), cell-conditioned medium, and proteoglycans in mediating retrovirus binding to FN. We also examined the role of Polybrene (PB) in these interactions. We found that gp70 is not involved in retrovirus binding to FN. Immobilization of the virus, however, does not overcome its receptor requirement, and gp70 is still needed for successful gene transfer. Our results clearly show that retrovirus binds FN through virus-associated heparan sulfate (HS) and that binding is necessary for transduction without PB. Two distinct modes of gene transfer occur depending on PB: (i) in the presence of PB, retrovirus interacts directly with the target cells; and (ii) in the absence of PB, retrovirus binds to FN and target cells interact with the immobilized virus. PB may promote the former mode by interacting with the virus HS and reducing the negative charge of the viral particles. Interestingly, the latter mode is more efficient, leading to significantly enhanced gene transfer. A better understanding of these interactions may provide insight into virus-cell interactions and lead to a more rational design of transduction protocols.