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Identification of genes involved in latency and reactivation of pseudorabiesvirus, use in biological containment study of viral genomes in pigs

Objective

The joint proposal aims at the identification of viral genes influencing establishment of and reactivation from latency of pseudorabies virus (PRV); Aujeszky's disease virus; herpes suid 1) in the natural host, swine. The major objective will be a detailed analysis on the biosafety of PRV mutants specifically manipulated in latency-associated (LA) functions.
The goal of the project is the identification of viral genes influencing the establishment of and reactivation from latency of pseudorabies virus (PRV). The major objective is a detailed analysis on the biosafety of PRV mutants specifically manipulated in latency associated (LA) functions. LA transcripts (LAT) will be identified, characterized, and the molecular organization of this genomic region will be elucidated. Neural cell cultures will be produced and invitro assays of those porcine neural cells which reflect a natural target cell of PRV can be studied invivo on pigs to analyse their latency characteristics.

Work to date includes:
characterization of strains invivo;
analysis of LAT region of PRV;
analysis of viral gene expression of lymphocytes from latently infected pigs;
an in vitro study of Aujeszky virus interactions with cultured cells from the nervous system;
characterization of messenger ribonucleic acids (mRNA) expressed in tissues of latently infected pigs;
construction of a complementary deoxyribonucleic acid (cDNA) bank.

The project is continuing.
LA transcripts (LAT's) will be identified, characterized, and the molecular biological studies (in situ hybridization, Northern blotting, protection assay, PCR, cDNA bank) will be done in a convergent and complementary way between the different laboratories. The results will enable construction of viable PRV mutants. After in vitro assays, those mutants can be immediately investigated in pigs to analyse their organ tropism and latency characteristics. In vitro culture systems of porcine neural cells reflecting a natural target cell of PRV shall be established. Finally, identification of PRV gene(s) specific for latency would offer for the first time the availability of a latency-marker in PRV-infection, and its applicability for different diagnostic purposes will be tested. Sensitive detection of LAT (eg by PCR) or of associated peptides (eg by serology) could unambiguously identify latently infected pigs carrying reactivable PRV strains or live vaccines.

Coordinator

Centre National d'Études Vétérinaires et Alimentaires (CNEVA)
Address
22 Rue Pierre Curie
94701 Maisons-alfort
France

Participants (4)

Centre National de la Recherche Scientifique (CNRS)
France
Address
7 Rue Guy Môquet
94802 Villejuif
FRIEDRICH-LOEFLER INSTITUTE FEDERAL RESEARCH INSTITUT FOR ANIMAL HEALTH
Germany
Address
Boddenblick 5 A
17498 Insel Riems
LABORATORIO NACIONAL DE INVESTIGACAO VETERINARIA
Portugal
Address
701,Estrada De Benfica 701
1549 Lisboa
Rhône Merieux SA
France
Address
254 Rue Marcel Merieux
69342 Lyon