Molecularly engineered resistance to California serogroup virus replication in mosquito cells and mosquitoes.
Introduction of genetic elements derived from a viral pathogen's genome may be used to reduce the vectorial capacity of mosquitoes for that virus. A double subgenomic Sindbis virus expression system was utilized to transcribe sequences of LaCrosse (LAC) virus small (S) or medium (M) segment RNA in sense or antisense orientation; wild-type Sindbis and LaCrosse viruses have single-stranded RNA genomes, the former being positive sense and the latter being negative sense. Recombinant viruses were generated and used to infect Aedes albopictus (C6/36) mosquito cells, which were challenged with wild-type LAC virus and then assayed for LAC virus replication. Several recombinant viruses containing portions of the LAC S segment were capable of inducing varying degrees of interference to the challenge virus. Cells infected with TE/3'2J/ANTI-S virus, expressing full-length negative-sense S RNA of LAC virus, yielded 3-6 log10TCID50 (tissue culture 50% infective dose) less LAC virus per ml than did cells infected with a double subgenomic sindbis virus containing no LAC insert. When C6/36 cells infected with TE/3'2J/ANTI-S were challenged with closely related heterologous bunyaviruses, a similar inhibitory effect was seen. Adult Ae. triseriatus mosquitoes infected with TE/3'2J/ANTI-S were also resistant to challenge by LAC virus. Organs that were productively infected by the double subgenomic Sindbis virus expressing the LAC anti-S sequences demonstrated little LAC virus or antigen. These studies indicate that expression of carefully selected antiviral sequences derived from the pathogen's genome may result in efficacious molecular viral interference in mosquito cells and, more importantly, in mosquitoes.
Powers
A M
AM
Department of Microbiology, Colorado State University, Ft. Collins 80523, USA.
Kamrud
K I
KI
Olson
K E
KE
Higgs
S
S
Carlson
J O
JO
Beaty
B J
BJ
D000330:Aedes / Q000821:virology*; D000818:Animals; D000956:Antigens, Viral / Q000032:analysis; D015152:Blotting, Northern; D002043:Bunyaviridae / Q000502:physiology; D002460:Cell Line; D004198:Disease Susceptibility; D005822:Genetic Vectors; D016678:Genome; D018062:La Crosse virus / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology*; D016297:Mutagenesis, Site-Directed; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000096:biosynthesis; D012845:Sindbis Virus; D013997:Time Factors; D014158:Transcription, Genetic; D014779:Virus Replication
D000956:Antigens, Viral; D012367:RNA, Viral