Haas-Stapleton Eric J, Washburn Jan O, Volkman Loy E
Department of Plant and Microbial Biology, 251 Koshland Hall, University of California, Berkeley, CA 94720-3102, USA.
J Gen Virol. 2005 May;86(Pt 5):1349-1355. doi: 10.1099/vir.0.80845-0.
Spodoptera frugiperda larvae are highly resistant to oral infection by Autographa californica multiple nucleopolyhedrovirus (AcMNPV) (LD(50), approximately 9200 occlusions), but extremely susceptible to budded virus within the haemocoel (LD(50), <1 p.f.u.). The inability of AcMNPV occlusion-derived virus (ODV) to establish primary infections readily within midgut cells accounts for a major proportion of oral resistance. To determine whether inappropriate binding of AcMNPV ODV to S. frugiperda midgut cells contributes to lack of oral infectivity, the binding and fusion properties of AcMNPV ODV were compared with those of the ODV of a new isolate of Spodoptera frugiperda multiple nucleopolyhedrovirus (SfMNPV) obtained from a field-collected larva (oral LD(50), 12 occlusions). By using a fluorescence-dequenching assay conducted in vivo, it was found that AcMNPV ODV bound to the midgut epithelia of S. frugiperda larvae at approximately 15 % of the level of SfMNPV ODV, but that, once bound, the efficiencies of fusion for the two ODVs were similar: 60 % for AcMNPV and 53 % for SfMNPV. Whilst the difference in binding efficiencies was significant, it could not account entirely for the observed differences in infectivity. Competition experiments, however, revealed that, in S. frugiperda larvae, SfMNPV ODV bound to a midgut cell receptor that was not bound by AcMNPV ODV, indicating that ODV interaction with a specific receptor(s) was necessary for productive infection of midgut columnar epithelial cells. Fusion in the absence of this ligand-receptor interaction did not result in productive infections.
草地贪夜蛾幼虫对苜蓿银纹夜蛾多核多角体病毒(AcMNPV)的口服感染具有高度抗性(半数致死剂量,约9200个包涵体),但对血腔中的出芽病毒极为敏感(半数致死剂量,<1个空斑形成单位)。AcMNPV包涵体衍生病毒(ODV)无法在中肠细胞内轻易建立原发性感染,这是口服抗性的主要原因。为了确定AcMNPV ODV与草地贪夜蛾中肠细胞的不适当结合是否导致口服感染性缺乏,将AcMNPV ODV的结合和融合特性与从田间采集的幼虫中获得的草地贪夜蛾多核多角体病毒(SfMNPV)新分离株的ODV进行了比较(口服半数致死剂量,12个包涵体)。通过体内荧光猝灭试验发现,AcMNPV ODV与草地贪夜蛾幼虫中肠上皮的结合水平约为SfMNPV ODV的15%,但一旦结合,两种ODV的融合效率相似:AcMNPV为60%,SfMNPV为53%。虽然结合效率的差异很显著,但它不能完全解释观察到的感染性差异。然而,竞争实验表明,在草地贪夜蛾幼虫中,SfMNPV ODV与AcMNPV ODV未结合的中肠细胞受体结合,这表明ODV与特定受体的相互作用是中肠柱状上皮细胞有效感染所必需的。在没有这种配体-受体相互作用的情况下发生融合不会导致有效感染。