INTRACELLULAR IMMUNIZATION OF MOSQUITO CELLS TO LACROSSE VIRUS USING A RECOMBINANT SINDBIS VIRUS VECTOR

被引:39
作者
POWERS, AM [1 ]
OLSON, KE [1 ]
HIGGS, S [1 ]
CARLSON, JO [1 ]
BEATY, BJ [1 ]
机构
[1] COLORADO STATE UNIV, DEPT MICROBIOL, ARTHROPOD BORNE ANIM DIS RES LAB, FT COLLINS, CO 80523 USA
关键词
SINDBIS; LACROSSE; INTERFERENCE; ANTISENSE; RNA VIRUS;
D O I
10.1016/0168-1702(94)90061-2
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
A cDNA of the small RNA genome segment of La Crosse (LAC) virus was inserted, in an antisense orientation, into a double subgenomic Sindbis (dsSIN) virus expression vector generating pTE/3'2J/ANTI-S (15,000bp). In vitro transcription of the pTE/3'2J/ANTI-S template generated genomic RNA that was electrotransfected into BHK-21 cells to produce virus. Northern blot analysis of RNA isolated from infected Aedes albopictus (C6/36) cells showed that the TE/3'2J/ANTI-S virus produced a subgenomic mRNA of the appropriate size, indicating transcription of the LAC cDNA segment. C6/36 cells were infected with either TE/3'2J/ANTI-S, TE/3'2J (a dsSIN virus with no LAC insert), or wild type Sindbis (SIN, strain AR339) viruses and subsequently challenged with LAC virus. LAC virus titers were determined using a capture antibody ELISA. Mosquito cells infected with TE/3'2J/ ANTI-S virus yielded at least 4 log(10) TCID50/ml less LAC virus than cells infected with either TE/3'2J or AR339 SIN viruses. The use of the infectious SIN virus expression vectors provides a novel approach for high level cytoplasmic expression of genes or sequences of interest in arthropod cells, and for evaluating strategies for intracellular immunization against arboviruses.
引用
收藏
页码:57 / 67
页数:11
相关论文
共 41 条
[1]  
BEATY BJ, 1991, CURR TOP MICROBIOL, V169, P27
[2]  
CHOMCZYNSKI P, 1987, ANAL BIOCHEM, V162, P156, DOI 10.1016/0003-2697(87)90021-2
[3]   VIRAL PROTECTION IN TRANSGENIC TOBACCO PLANTS EXPRESSING THE CUCUMBER MOSAIC-VIRUS COAT PROTEIN OR ITS ANTISENSE RNA [J].
CUOZZO, M ;
OCONNELL, KM ;
KANIEWSKI, W ;
FANG, RX ;
CHUA, NH ;
TUMER, NE .
BIO-TECHNOLOGY, 1988, 6 (05) :549-&
[4]   CHARACTERIZATION OF RNA-MEDIATED RESISTANCE TO TOMATO SPOTTED WILT VIRUS IN TRANSGENIC TOBACCO PLANTS [J].
DEHAAN, P ;
GIELEN, JJL ;
PRINS, M ;
WIJKAMP, IG ;
VANSCHEPEN, A ;
PETERS, D ;
VANGRINSVEN, MQJM ;
GOLDBACH, R .
BIO-TECHNOLOGY, 1992, 10 (10) :1133-1137
[5]   A STABLE BIFUNCTIONAL ANTISENSE TRANSCRIPT INHIBITING GENE-EXPRESSION IN TRANSGENIC PLANTS [J].
DELAUNEY, AJ ;
TABAEIZADEH, Z ;
VERMA, DPS .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1988, 85 (12) :4300-4304
[6]   ROLE OF RETICULOENDOTHELIOSIS VIRUS ENVELOPE GLYCOPROTEIN IN SUPERINFECTION INTERFERENCE [J].
DELWART, EL ;
PANGANIBAN, AT .
JOURNAL OF VIROLOGY, 1989, 63 (01) :273-280
[7]   PERSISTENT INFECTION OF AEDES-ALBOPICTUS C6/36 CELLS BY BUNYAMWERA VIRUS [J].
ELLIOTT, RM ;
WILKIE, ML .
VIROLOGY, 1986, 150 (01) :21-32
[8]   MOLECULAR-BIOLOGY OF THE BUNYAVIRIDAE [J].
ELLIOTT, RM .
JOURNAL OF GENERAL VIROLOGY, 1990, 71 :501-522
[9]   DNA-MEDIATED GENE-TRANSFER - APPLICATIONS TO MOSQUITOS [J].
FALLON, AM .
NATURE, 1991, 352 (6338) :828-829
[10]   EXPRESSION OF AVIAN RETICULOENDOTHELIOSIS VIRUS ENVELOPE CONFERS HOST-RESISTANCE [J].
FEDERSPIEL, MJ ;
CRITTENDEN, LB ;
HUGHES, SH .
VIROLOGY, 1989, 173 (01) :167-177