Ramalho-Santos J, de Lima M C
Center for Neuroscience and Cell Biology of Coimbra, University of Coimbra, Portugal.
Biosci Rep. 2001 Jun;21(3):293-304. doi: 10.1023/a:1013282015121.
We have directly compared the effect of two types of dextran sulfate with distinct molecular weights (500 kDa and 5 kDa) on the fusion activity and infectivity of both Sendai and influenza viruses, two lipid-enveloped viruses that differ in their routes of entry into target cells. To correlate membrane merging and infectivity MDCK cells were used as targets for the viruses in both approaches. In either case pronounced inhibition of virus-cell interactions by dextran sulfate was only observed at low pH, even though Sendai virus fuses maximally at pH 7.4. Although membrane merging could not be fully abolished, the inhibitory effect was always greater when the higher molecular weight dextran sulfate was used. The presence of this residual fusion activity, that could not be reduced even with high concentrations of agent, suggests that a limited number of binding sites for dextran sulfate may exist on the viral envelopes. The compounds also inhibited fusion of bound virions, and all results could be reproduced using erythrocyte ghosts as target membranes in the fusion assay, instead of MDCK cells. In agreement with these observations only the infectivity of influenza virus (which requires a low pH-dependent step to enter target cells) was affected by dextran sulfate, again the higher molecular weight compound showing a more pronounced inhibitory effect.
我们直接比较了两种分子量不同(500 kDa和5 kDa)的硫酸葡聚糖对仙台病毒和流感病毒的融合活性及感染性的影响,这两种包膜病毒进入靶细胞的途径不同。为了关联膜融合与感染性,在两种方法中均使用MDCK细胞作为病毒的靶细胞。在任何一种情况下,仅在低pH值下才观察到硫酸葡聚糖对病毒 - 细胞相互作用有明显抑制作用,尽管仙台病毒在pH 7.4时融合能力最强。虽然膜融合不能完全消除,但使用较高分子量的硫酸葡聚糖时抑制作用总是更大。即使使用高浓度试剂也无法降低这种残余融合活性的存在,表明病毒包膜上可能存在有限数量的硫酸葡聚糖结合位点。这些化合物还抑制结合病毒粒子的融合,并且在融合试验中使用红细胞血影作为靶膜代替MDCK细胞时,所有结果均可重现。与这些观察结果一致,只有流感病毒的感染性(其进入靶细胞需要低pH依赖性步骤)受到硫酸葡聚糖的影响,同样,较高分子量的化合物显示出更明显的抑制作用。