PMID- 11118361
OWN - NLM
STAT- MEDLINE
DCOM- 20010126
LR  - 20061115
IS  - 0042-6822 (Print)
IS  - 0042-6822 (Linking)
VI  - 278
IP  - 2
DP  - 2000 Dec 20
TI  - Expression of a foreign epitope by porcine reproductive and respiratory syndrome 
      virus.
PG  - 380-9
AB  - The potential of porcine reproductive and respiratory syndrome virus (PRRSV) as a 
      viral vector was explored by the insertion of a sequence encoding a foreign 
      antigen into the infectious cDNA clone of the Lelystad virus isolate. An epitope 
      of the hemagglutinin (HA) protein of human influenza A virus was introduced at 
      the 5' end and at the 3' end of ORF7, in each case resulting in a fusion protein 
      between the HA epitope and the nucleocapsid (N) protein. Furthermore, in the 
      construct carrying the HA sequences at the 5' end of ORF7, additional in-frame 
      insertions encoding the autoprotease 2A of foot-and-mouth disease virus were made 
      between the HA and ORF7 sequences to ensure the generation of a functional N 
      protein from its hybrid precursor. When RNA transcripts from these full-length 
      cDNA clones were transfected into BHK-21 cells, they were each found to 
      replicate, to express the HA epitope, and to produce progeny virus. However, 
      fusion of the HA epitope directly to the nucleocapsid protein either at the N 
      terminus or at the C terminus adversely affected both the viability and the 
      genetic stability of the recombinant PRRS viruses. Serial passage of the 
      recombinant viruses on porcine alveolar macrophages demonstrated that these 
      viruses had lost (part of) the HA epitope at passage four. In contrast, in the 
      PRRS viruses expressing the HA epitope from a precursor cleavable by the 
      autoprotease 2A peptide, the HA epitope was still intact after four passages, and 
      no effect on the viability of these viruses was observed. Immunoprecipitation and 
      pulse chase experiments revealed the efficient and presumably cotranslational 
      cleavage of the HA epitope from the N protein by the 2A protease. Our results 
      demonstrate the feasibility of using PRRSV as a viral vector that might be 
      suitable for the delivery of antigens from other pathogens to the immune system 
      of the pig.
CI  - Copyright 2000 Academic Press.
FAU - Groot Bramel-Verheije, M H
AU  - Groot Bramel-Verheije MH
AD  - Department of Mammalian Virology, Institute for Animal Science and Health, 
      Lelystad, NL-8200 AB, The Netherlands. m.h.grootbramel-verheije@id.wag-ur.nl
FAU - Rottier, P J
AU  - Rottier PJ
FAU - Meulenberg, J J
AU  - Meulenberg JJ
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Virology
JT  - Virology
JID - 0110674
RN  - 0 (DNA Primers)
RN  - 0 (Epitopes)
RN  - 0 (Hemagglutinin Glycoproteins, Influenza Virus)
RN  - 0 (Recombinant Proteins)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Base Sequence
MH  - Cell Line
MH  - Cloning, Molecular
MH  - Cricetinae
MH  - DNA Primers
MH  - Epitopes/chemistry/*genetics
MH  - *Genetic Vectors
MH  - Hemagglutinin Glycoproteins, Influenza Virus/chemistry/*genetics
MH  - Humans
MH  - Influenza A virus/*genetics
MH  - Kinetics
MH  - Molecular Sequence Data
MH  - Mutagenesis, Site-Directed
MH  - Nucleocapsid/genetics
MH  - Open Reading Frames
MH  - Porcine respiratory and reproductive syndrome virus/*genetics
MH  - Recombinant Proteins/biosynthesis/chemistry
MH  - Swine
MH  - Transcription, Genetic
MH  - Transfection
EDAT- 2000/12/19 11:00
MHDA- 2001/02/28 10:01
CRDT- 2000/12/19 11:00
PHST- 2000/12/19 11:00 [pubmed]
PHST- 2001/02/28 10:01 [medline]
PHST- 2000/12/19 11:00 [entrez]
AID - S0042-6822(00)90525-7 [pii]
AID - 10.1006/viro.2000.0525 [doi]
PST - ppublish
SO  - Virology. 2000 Dec 20;278(2):380-9. doi: 10.1006/viro.2000.0525.