Cellular permissivity factor for viruses and uses thereof
View Patent ↗The present invention provides methods and compositions related to the generation of host cells permissive for virus growth, particularly Porcine Reproductive and Respiratory Syndrome (PRRS) virus.
1. A method of facilitating production of Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) from a culture of vertebrate cells, comprising the steps of:
(a) providing a recombinant vertebrate cell transfected with an exogenous polynucleotide that encodes a polypeptide having a transmembrane domain and at least 85% sequence identity to SEQ ID NO:2, so that expression of the polypeptide in said cell is increased;
(b) contacting a culture of said cell with PRRSV virus under conditions which permit infection of the cells and growth of the virus; and
(c) recovering virus from said culture.
2. The method of claim 1 , wherein the cell was previously PRRSV permissive and is rendered more PRRSV permissive.
3. The method of claim 1 , wherein the cell is selected from the group consisting of baby hamster kidney cells (BHK21), porcine kidney cells, feline kidney cells, avian cells, and swine testicular cells.
4. The method of claim 1 , wherein the PRRSV is of the European genotype.
5. The method of claim 1 , wherein the PRRSV is of the North American genotype.
6. The method of claim 1 , wherein the polynucleotide encodes a polypeptide having a transmembrane domain, and at least 90% sequence identity to SEQ ID NO:2.
7. The method of claim 1 , wherein the polynucleotide encodes a polypeptide having a transmembrane domain and at least 95% sequence identity to SEQ ID NO:2.
8. The method of claim 1 , wherein the polynucleotide encodes a polypeptide having a transmembrane domain and at least 99% sequence identity to SEQ ID NO:2.
9. The method of claim 1 , further comprising the step of producing a vaccine from the recovered virus.
10. The method of claim 9 , wherein the vaccine comprises inactivated virus.
11. The method of claim 9 , wherein the vaccine comprises live attenuated virus.
12. The method of claim 1 , wherein transfection with the exogenous polynucleotide is accomplished by electroporation.
13. The method of claim 1 , wherein said exogenous polynucleotide encodes a fusion protein.