Recombinant attenuated parvovirus
The invention is in the field of virus vaccines for protecting animals against infection by parvovirus, their production and use. More in particular, the invention is related to a vaccine comprising an attenuated parvovirus comprising a capsid protein or fragment thereof derivable from another parvovirus. Surprisingly it was found that such a vaccine was capable of inducing higher titers of protecting antibodies against a challenge with the second type of parvovirus strains while maintaining good immunity against the first type of parvovirus. Recombinant virus strains also remained attenuated.
1. An attenuated recombinant parvovirus comprising a DNA sequence obtained from an attenuated first parvovirus;
wherein the DNA sequence encoding the capsid protein or fragment thereof of said first parvovirus is replaced by a DNA sequence encoding a capsid protein or fragment thereof derived from a second parvovirus;
wherein said first parvovirus is a type 2 canine parvovirus;
wherein said second parvovirus is a type 2c canine parvovirus;
wherein the attenuated recombinant canine parvovirus comprises the DNA sequence encoding the capsid protein of a canine parvovirus type 2c isolate within the DNA sequence of an attenuated canine parvovirus type 2 virus; and wherein said fragment thereof of said second parvovirus comprises at least one amino acid difference from said fragment thereof of said first parvovirus.
2. The attenuated recombinant parvovirus according to claim 1 wherein the DNA sequence obtained from an attenuated first parvovirus is essentially full length.
3. The attenuated recombinant parvovirus according to claim 2 , wherein said capsid protein from said second parvovirus is essentially full-length.
4. A vaccine for the protection of animals against infection with parvovirus, wherein said vaccine comprises the attenuated recombinant parvovirus according to claim 3 and a pharmaceutically acceptable carrier.
5. A method for obtaining a recombinant parvovirus according to claim 3 , said method comprising the steps of:
a. Obtaining at least one DNA fragment from an attenuated first parvovirus strain not encoding a capsid protein;
b. Obtaining at least one DNA fragment from a second parvovirus strain encoding a capsid protein;
c. Transfecting a cell permissive for parvovirus with the DNA fragments obtained in steps a and b;
d. Allowing the DNA fragments to recombine;
e. Selecting an attenuated recombinant virus that encodes a viral capsid protein derived from the second parvovirus in the genetic context of the genome of the first parvovirus;
f Culturing the cell under conditions that allow the production of parvovirus in a cell culture; and
g. Obtaining the recombinant parvovirus from the cell culture.
6. The attenuated recombinant parvovirus according to claim 1 , wherein said capsid protein from said second parvovirus is essentially full-length.
7. A vaccine for the protection of animals against infection with parvovirus, wherein said vaccine comprises the attenuated recombinant parvovirus according to claim 6 and a pharmaceutically acceptable carrier.
8. A vaccine for the protection of animals against infection with parvovirus, wherein said vaccine comprises the attenuated recombinant parvovirus according to claim 2 and a pharmaceutically acceptable carrier.
9. A vaccine for the protection of animals against infection with parvovirus, wherein said vaccine comprises an attenuated recombinant parvovirus according to claim 1 and a pharmaceutically acceptable carrier.
10. A method for obtaining a recombinant parvovirus according to claim 6 , said method comprising the steps of:
a. Obtaining at least one DNA fragment from an attenuated first parvovirus strain not encoding a capsid protein;
b. Obtaining at least one DNA fragment from a second parvovirus strain encoding a capsid protein;
c. Transfecting a cell permissive for parvovirus with the DNA fragments obtained in steps a and b;
d. Allowing the DNA fragments to recombine;
e. Selecting an attenuated recombinant virus that encodes a viral capsid protein derived from the second parvovirus in the genetic context of the genome of the first parvovirus;
f Culturing the cell under conditions that allow the production of parvovirus in a cell culture; and
g. Obtaining the recombinant parvovirus from the cell culture.
11. A method for obtaining a recombinant parvovirus according to claim 2 , said method comprising the steps of:
a. Obtaining at least one DNA fragment from an attenuated first parvovirus strain not encoding a capsid protein;
b. Obtaining at least one DNA fragment from a second parvovirus strain encoding a capsid protein;
c. Transfecting a cell permissive for parvovirus with the DNA fragments obtained in steps a and b;
d. Allowing the DNA fragments to recombine;
e. Selecting an attenuated recombinant virus that encodes a viral capsid protein derived from the second parvovirus in the genetic context of the genome of the first parvovirus;
f. Culturing the cell under conditions that allow the production of parvovirus in a cell culture; and
g. Obtaining the recombinant parvovirus from the cell culture.
12. A method for obtaining a recombinant parvovirus according to claim 1 , said method comprising the steps of:
a. Obtaining at least one DNA fragment from an attenuated first parvovirus strain not encoding a capsid protein;
b. Obtaining at least one DNA fragment from a second parvovirus strain encoding a capsid protein;
c. Transfecting a cell permissive for parvovirus with the DNA fragments obtained in steps a and b;
d. Allowing the DNA fragments to recombine;
e. Selecting an attenuated recombinant virus that encodes a viral capsid protein derived from the second parvovirus in the genetic context of the genome of the first parvovirus;
f. Culturing the cell under conditions that allow the production of parvovirus in a cell culture; and
g. Obtaining the recombinant parvovirus from the cell culture.
13. A method of protecting an animal against infection with canine parvovirus comprising administering to the animal the vaccine of claim 9 .
14. A method of protecting an animal against infection with canine parvovirus comprising administering to the animal the vaccine of claim 8 .
15. A method of protecting an animal against infection with canine parvovirus comprising administering to the animal the vaccine of claim 7 .
16. A method of protecting an animal against infection with canine parvovirus comprising administering to the animal the vaccine of claim 4 .