Engineered resistance in Aedes aegypti to a West African and a South American strain of yellow fever virus
Engineered resistance in Aedes aegypti to a West African and a South American strain of yellow fever virus
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DOI：
10.4269/ajtmh.1998.58.663
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发表时间：
1998-05-01
影响因子：
3.3
通讯作者：
Blair, CD
中科院分区：
文献类型：
--
作者：
Higgs, S;Rayner, JO;Blair, CD
利用黄热病病毒（YFV）17 D疫苗株的前膜（prM）或聚合酶（NS 5）编码区的反义RNA，将双亚基因组辛德毕斯（dsSIN）病毒工程化到蚊子细胞中。用每种dsSIN病毒以高感染复数（MOI）感染白纹伊蚊C6/36细胞。48小时后，用MOI为0.1的YFV Asibi株攻击转导的细胞。在攻击后72小时，通过免疫荧光法测定细胞中是否存在YFV抗原。用来自YFV基因组的prM或NS 5反义RNA转导的细胞不显示YFV特异性抗原。相反，用不表达反义RNA或登革病毒衍生的反义RNA的对照dsSIN病毒感染的细胞对攻击病毒是允许的。为了分析蚊子中的抗性，将每种dsSIN病毒的五个log（10）50%组织培养感染剂量（TCID 50）和野生型YFV的西非（BA-55）或南美（1899/81）毒株的三个log（10）TCID（50）共接种到Ae.埃及人。用靶向prM或NS 5基因区域的效应子RNA转导的蚊子不传播西非YFV，并且传播YFV的南美毒株很差。
Double subgenomic Sindbis (dsSIN) viruses were engineered to transduce mosquito cells with antisense RNA derived either from the premembrane (prM) or polymerase (NS5) coding regions of the 17D vaccine strain of yellow fever virus (YFV). Aedes albopictus C6/36 cells were infected at high multiplicities of infection (MOI) with each dsSIN virus. Forty-eight hours later, the transduced cells were challenged with an MOI of 0.1 of the Asibi strain of YFV. At 72-hr postchallenge, the cells were assayed by immunofluorescence for the presence of YFV antigen. Cells transduced with prM or NS5 antisense RNAs derived from the YFV genome displayed no YFV-specific antigens. In contrast, cells infected with control dsSIN viruses that expressed no antisense RNA or dengue virus-derived antisense RNAs were permissive for the challenge virus. To analyze resistance in the mosquito, five log(10) 50% tissue culture infective doses (TCID50) of each dsSIN virus and three log(10)TCID(50) of either a West African (BA-55) or South American (1899/81) strain of wild-type YFV were coinoculated into Ae. aegypti. Mosquitoes transduced with effector RNAs targeting the prM or NS5 gene regions did not transmit West African YFV and poorly transmitted the South American strain of YFV.