Live attenuated parvovirus
The invention relates to live attenuated parvoviruses, their uses, vaccines comprising such live attenuated parvoviruses, as well as methods for their production.
1. A live attenuated canine parvovirus type 2 (CPV2), wherein said CPV2 comprises a capsid gene coding for both an amino acid other than isoleucine at amino acid position 219 of the capsid protein and an amino acid other than glutamine at amino acid position 386 of the capsid protein;
wherein a DNA fragment of a part of the non-capsid region of said CPV2 is replaced by a homologous DNA fragment of a part of the non-capsid region derived from a second parvovirus, wherein said second parvovirus is a canine parvovirus 2, and wherein said homologous DNA fragment of said second parvovirus carries an attenuating mutation in the nonstructural region in the region from position 2061 to 2072, due to the replacement of the DNA sequence of wild-type CPV2 in the region from position 2061 to 2072 encoding the amino acid sequence LTPL (SEQ ID NO: 6) with a DNA sequence encoding the amino acid sequence HVRM (SEQ ID NO: 7).
2. A vaccine for protecting an animal against parvovirus, comprising the live attenuated CPV2 of claim 1 and a pharmaceutically acceptable carrier.
3. A method for treating an animal comprising administering to said animal the live attenuated CPV2 of claim 1 .
4. The live attenuated CPV2 according to claim 1 that encodes a capsid protein of CPV2 serotype 2a, 2b, or 2c, or a capsid protein from feline parvovirus.
5. A vaccine for protecting an animal against parvovirus, comprising the live attenuated CPV2 of claim 4 and a pharmaceutically acceptable carrier.
6. A combination vaccine for the protection of animals against pathogens, characterized in that said combination vaccine comprises a vaccine according to claim 5 and an additional antigen of a virus or micro-organism pathogenic to animals or genetic information encoding an immunogenic protein of said virus or micro-organism.
7. A combination vaccine according to claim 6 , characterized in that said virus or micro-organism pathogenic to animals is selected from the group consisting of Ehrlichia canis, Babesia gibsoni, vogeli, rossi, Leishmania donovani -complex, Canine adenovirus, Canine coronavirus, Canine distempervirus, Leptospira interrogans serovar canicola, icterohaemorrhagiae, pomona, grippotyphosa, Bratislava , Canine hepatitisvirus, Canine parainfluenzavirus, rabies virus, Hepatozoon canis, Borrelia burgdorferi, Bordetella bronchiseptica , feline Herpesvirus, feline calicivirus, feline panleucopenia and Chlamydophila felis.
8. A method for making the vaccine of claim 5 , comprising mixing the live attenuated CPV2 with a pharmaceutically acceptable carrier.
9. A method of protecting an animal against parvovirus infection comprising administering to the animal the vaccine of claim 5 .
10. The live attenuated CPV2 according to claim 1 , wherein said CPV2 comprises a capsid gene coding for a valine at amino acid position 219 of the capsid protein, a lysine at amino acid position 386 of the capsid protein, or both a valine at amino acid position 219 of the capsid protein and a lysine at amino acid position 386 of the capsid protein.
11. A vaccine for protecting an animal against parvovirus, comprising the live attenuated CPV2 of claim 10 and a pharmaceutically acceptable carrier.
12. The live attenuated CPV2 according to claim 10 , wherein said CPV2 comprises a capsid gene coding for a valine at amino acid position 219 of the capsid protein and a lysine at amino acid position 386 of the capsid protein.
13. A vaccine for protecting an animal against parvovirus, comprising the live attenuated CPV2 of claim 12 and a pharmaceutically acceptable carrier.
14. A method of using recombinant DNA techniques to make the live attenuated CPV2 of claim 12 comprising altering both a codon of the capsid gene that encodes the isoleucine at amino acid position 219 to encode an amino acid other than isoleucine and a codon of the capsid gene that encodes the glutamine at amino acid position 386 to encode an amino acid other than glutamine.
15. A method for preparing the live attenuated CPV2 of claim 12 comprising both exchanging a DNA fragment encoding at least part of the CPV2 capsid protein comprising a codon encoding isoleucine at amino acid position 219 with a DNA fragment encoding the same part of the capsid protein comprising a codon encoding an amino acid other than isoleucine at amino acid position 219, and exchanging a DNA fragment encoding at least part of the CPV2 capsid protein comprising a codon encoding glutamine at amino acid position 386 with a DNA fragment encoding the same part of the capsid protein comprising a codon encoding an amino acid other than glutamine at amino acid position 386.
16. The live attenuated CPV2 according to claim 12 that encodes a capsid protein of CPV2 serotype 2a, 2b, or 2c, or a capsid protein from feline parvovirus.
17. A vaccine for protecting an animal against parvovirus, comprising the live attenuated CPV2 of claim 16 and a pharmaceutically acceptable carrier.