Pathogen-derived resistance to dengue type virus 2 in mosquito cells by expression of the premembrane coding region of the viral genome
Pathogen-derived resistance to dengue type virus 2 in mosquito cells by expression of the premembrane coding region of the viral genome
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DOI：
10.1128/jvi.70.4.2132-2137.1996
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发表时间：
1996-04-01
影响因子：
5.4
通讯作者：
Blair, CD
中科院分区：
文献类型：
--
作者：
Gaines, PJ;Olson, KE;Blair, CD
登革病毒2型（DEN-2; Jamaica）基因组的全长前膜（prM）编码区在C6/36（白纹伊蚊）细胞中以来自双亚基因组辛德毕斯（dsSIN）病毒的有义或反义方向表达。北方（RNA）印迹分析证实感染的C6/36细胞中有正义或反义DEN-2 prM RNA的表达。通过免疫荧光测定，在用表达DEN-2正义RNA的dsSIN病毒感染的细胞中证明了PrM蛋白。用每种dsSIN病毒以50的感染复数（MOI）感染C6/36细胞，48小时后用DEN-2病毒以0.1的MOI攻击。尽管用对照dsSIN病毒感染的C6/36细胞支持高水平的DEN-2复制，但用表达prM反义RNA的dsSIN病毒感染的C6/36细胞对DEN-2攻击具有完全抗性。表达prM蛋白或不可翻译的prM正义RNA的细胞也对DEN-2攻击具有抗性。表达prM蛋白的细胞在MOI为10时表现出DEN-2病毒的一些突破。然而，表达的不可翻译的正义prM RNA在高MOI下赋予对攻击的完全保护。
The full-length premembrane (prM) coding region of the dengue virus type 2 (DEN-2; Jamaica) genome was expressed in C6/36 (Adedes albopictus) cells in either the sense or the antisense orientation from a double subgenomic Sindbis (dsSIN) virus. Northern (RNA) blot analysis confirmed the expression of sense or antisense DEN-2 prM RNA in infected C6/36 cells. PrM protein was demonstrated in cells infected with dsSIN viruses expressing DEN-2 sense RNAs by an immunofluorescence assay. C6/36 cells were infected with each dsSIN virus at a multiplicity of infection (MOI) of 50 and challenged 48 h later with DEN-2 virus at an MOI of 0.1. Whereas C6/36 cells infected with a control dsSIN virus supported high levels of DEN-2 replication, C6/36 cells infected with the dsSIN virus expressing prM antisense RNA were completely resistant to DEN-2 challenge. Cells expressing prM protein or untranslatable prM sense RNA also were resistant to DEN-2 challenge. Cells expressing prM protein demonstrated some breakthrough of DEN-2 virus when challenged at an MOI of 10. However, expressed untranslatable sense prM RNA conferred complete protection to challenge at the high MOI.