recombinant poxviruses with foreign DNA in essential regions

• US 5,766,882 A
• Filed: 03/14/1996
• Issued: 06/16/1998
• Est. Priority Date: 04/29/1994
• Status: Expired due to Fees

• Current Assignee: Immuno Aktiengesellschaft
• Original Assignee: Immuno Aktiengesellschaft
• Inventors: Holzer, Georg, Dorner, Friedrich, Falkner, Falko-Gunter
• Primary Examiner(s): Campell, Bruce R.
• Application Number:
  US08/616,133
• Time in Patent Office: 824 Days
• Field of Search: 435/320.1, 435/69.1, 435/70.3, 435/91.1, 435/91.42, 435/172.1, 435/172.3, 435/325, 435/349, 435/363, 435/366, 435/375, 514/44, 424/199.1, 424/232.1, 424/93.2, 536/24.1
• US Class Current: 435/69.1
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: A61K 39/00 Medicinal preparations containing antigens or antibodies materials for immunoassay G01N33/53
• CPC Class Codes: C07K 14/005 from viruses
• CPC Class Codes: C12N 15/86 Viral vectors
• CPC Class Codes: C12N 2710/24122 New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
• CPC Class Codes: C12N 2710/24143 viral genome or elements thereof as genetic vector
• CPC Class Codes: C12N 2740/16222 New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
• CPC Class Codes: C12N 2830/00 Vector systems having a special element relevant for transcription
• CPC Class Codes: C12N 2830/002 inducible enhancer/promoter combination, e.g. hypoxia, iron, transcription factor
• CPC Class Codes: C12N 2830/15 chimeric enhancer/promoter combination
• CPC Class Codes: C12N 2830/42 being an intron or intervening sequence for splicing and/or stability of RNA
• CPC Class Codes: C12N 2830/55 from bacteria
• CPC Class Codes: C12N 2830/60 from viruses
• CPC Class Codes: C12N 2840/203 having an IRES

Abstract

Defective poxviruses that lack a function imparted by an essential region of its parental poxvirus are provided for protein production and vaccination. A DNA polynucleotide encoding a protein is inserted into the defective poxvirus and placed under transcriptional control of a promoter. The defective poxvirus is viable when the lost function of the essential region is complemented by a host cell, transgenic animal or helper virus.

Claims

1. A defective poxvirus that lacks a function imparted by an essential region of its parental poxvirus, said defective poxvirus comprising a foreign DNA polynucleotide under transcriptional control of a promoter.

2. A defective poxvirus according to claim 1, wherein said promoter is operable with enzymes of said defective poxvirus.

3. A defective poxvirus according to claim 1, wherein said promoter is a poxvirus promoter.

4. A defective poxvirus according to claim 1, wherein said foreign DNA polynucleotide is inserted into said essential region.

5. A defective poxvirus according to claim 1, wherein said essential region has been deleted from said defective poxvirus.

6. A defective poxvirus according to claim 5, wherein a marker replaces said essential region.

7. A defective poxvirus according to claim 6, wherein said foreign DNA polynucleotide is inserted into said marker.

8. A defective poxvirus according to claim 1, wherein said parental poxvirus is an orthopoxvirus.

9. A defective poxvirus according to claim 1, wherein said parental poxvirus is vaccinia.

10. A defective poxvirus according to claim 9, wherein said essential region is an open reading frame selected from the group consisting of I7L, F18R, D13L, D6R, A8L, J1R, E7L, F11L, E4L, I1L, J3R, J4R, H7R, D4R and A6R.

24. A defective poxvirus according to claim 10, wherein the essential region is open reading frame D4R.

25. A defective poxvirus according to claim 1, wherein the foreign DNA polynucleotide is of animal origin.

26. A defective poxvirus according to claim 1, wherein the foreign DNA polynucleotide is of plant origin.

27. A defective poxvirus according to claim 1, wherein the foreign DNA polynucleotide is of bacterial origin.

28. A defective poxvirus according to claim 1, wherein the foreign DNA polynucleotide is of viral origin.

29. A defective poxvirus according to claim 1, wherein the foreign DNA polynucleotide is of fungal origin.

30. A defective poxvirus according to claim 1, wherein the foreign DNA polynucleotide is of protozoan origin.

31. A defective poxvirus according to claim 1, wherein the foreign DNA polynucleotide is synthetic.

32. A defective poxvirus according to claim 1, wherein the foreign DNA polynucleotide is a chimeric gene.

11. A cell line having the ability to complement a lost essential function of a defective poxvirus.

12. A cell line according to claim 11, wherein said ability is imparted by an essential region of a parental poxvirus of said defective poxvirus.

33. A cell line according to claim 12, wherein the cell line is based on RK 13 cells.

34. A cell line according to claim 12, wherein the cell line is based on CV-1 cells.

35. A cell line according to claim 12, wherein the cell line is based on Vero cells.

36. A cell line according to claim 12, wherein the cell line is based on 293 cells.

37. A cell line according to claim 12, wherein the cell line is based on MRC-5 cells.

38. A cell line according to claim 12, wherein the cell line is based on BSC-1 cells.

39. A cell line according to claim 12, wherein the cell line is based on LM (TK^-) cells.

40. A cell line according to claim 12, wherein the cell line is based on STO cells.

13. A method of producing a protein, comprising the steps of:

providing a defective poxvirus that lacks a function imparted by an essential region of its parental poxvirus, wherein said defective poxvirus comprises a foreign DNA polynucleotide encoding said protein and said DNA polynucleotide is under transcriptional control of a promoter;

infecting a complementing cell line with said defective poxvirus; and

harvesting said protein.

14. A method according to claim 13, wherein said promoter is operable with enzymes of said defective poxvirus.

15. A method according to claim 13, wherein said promoter is a poxvirus promoter.

16. A method according to claim 13, wherein said providing step is accomplished by inserting said DNA polynucleotide into said essential region of said parental poxvirus.

17. A method according to claim 13, wherein said providing step is accomplished by replacing said essential region of said parental poxvirus with said DNA polynucleotide.

18. A method according to claim 13, wherein said providing step is accomplished by replacing said essential region of said parental poxvirus with a marker.

19. A method according to claim 18, wherein said DNA polynucleotide is inserted into said marker.

20. A method according to claim 18, wherein said DNA polynucleotide replaces said marker.

21. A method according to claim 18, wherein said marker is a gpt gene.

22. A method according to claim 13, wherein said parental poxvirus is an orthopoxvirus.

23. A method according to claim 22, wherein said parental poxvirus is vaccinia.

41. A method according to claim 13, wherein the protein is from a pathogen.

42. A method according to claim 41, wherein the protein is from a bacterial, viral, fungal or protozoan source.

43. A method according to claim 13, wherein the cell line is based on RK 13 cells, CV-1 cells or Vero cells.

44. A method according to claim 13, wherein the cell line is based on 293, MRC-5 cells, BSC-1 cells, LM (TK^-) cells or STO cells.