Induction of a protective immune response in a mammal by injecting a DNA sequence

• US 6,214,804 B1
• Filed: 06/07/1995
• Issued: 04/10/2001
• Est. Priority Date: 03/21/1989
• Status: Expired due to Term

• Current Assignee: Vical, Inc. (Fresh Tracks Therapeutics, Inc.)
• Original Assignee: Vical, Inc. (Fresh Tracks Therapeutics, Inc.)
• Inventors: Carson, Dennis A., Felgner, Philip L., Malone, Robert Wallace, Wolff, Jon Asher, Rhodes, Gary H.
• Primary Examiner(s): Crouch, Deborah
• Application Number:
  US08/481,919
• Time in Patent Office: 2,134 Days
• Field of Search: 514/44, 435/455, 435/69.1
• US Class Current: 514/44.00R
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: A61K 9/1272 with substantial amounts of non-phosphatidyl, i.e. non-acylglycerophosphate, surfactants as bilayer-forming substances, e.g. cationic lipids with cholesterol as the only non-phosphatidyl surfactant A61K9/127; lipids as modifying agent A61K47/543
• CPC Class Codes: C07K 14/005 from viruses
• CPC Class Codes: C07K 14/61 Growth hormone [GH], i.e. somatotropin
• CPC Class Codes: C12N 15/87 Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
• CPC Class Codes: C12N 2740/16122 New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
• CPC Class Codes: C12N 9/0069 acting on single donors with incorporation of molecular oxygen, i.e. oxygenases (1.13)
• CPC Class Codes: C12N 9/1033 Chloramphenicol O-acetyltransferase (2.3.1.28)
• CPC Class Codes: C12N 9/1247 DNA-directed RNA polymerase (2.7.7.6)
• CPC Class Codes: C12N 9/78 acting on carbon to nitrogen bonds other than peptide bonds (3.5)
• CPC Class Codes: C12Y 113/12007 Photinus-luciferin 4-monooxygenase (ATP-hydrolysing) (1.13.12.7), i.e. firefly-luciferase

Abstract

A method for delivering an isolated polynucleotide to the interior of a cell in a vertebrate, comprising the interstitial introduction of an isolated polynucleotide into a tissue of the vertebrate where the polynucleotide is taken up by the cells of the tissue and exerts a therapeutic effect on the vertebrate. The method can be used to deliver a therapeutic polypeptide to the cells of the vertebrate, to provide an immune response upon in vivo translation of the polynucleotide, to deliver antisense polynucleotides, to deliver receptors to the cells of the vertebrate, or to provide transitory gene therapy.

Claims

1. A method of producing antibodies directed to gp-120 protein of HIV virus in a mouse, comprising injecting into muscle of said mouse a DNA plasmid constructed of sequence encoding said gp-120 operably linked to a CMV promoter in an amount sufficient to generate said antibodies;

wherein said plasmid is free from association with transfection-facilitating proteins, viral particles, liposomes, cationic lipids, and calcium phosphate precipitating agents.

2. A process for generating a detectable antibody response in a mammal, comprising:

administering in vivo into a tissue of the mammal a construct comprising a noninfectious, nonintegrating DNA sequence encoding an immunogen, wherein said DNA is operably linked to a promoter, in an amount sufficient that uptake of said construct into cells of the mammal occurs, and sufficient expression results, to generate the detectable antibody response,

wherein said construct is free from association with transfection-facilitating proteins, viral particles, liposomal formulations and charged lipids.

3. The process of claim 2, wherein said construct is introduced into mucous membrane tissue.

4. The process of claim 2, wherein the mammal is human.

5. The process of claim 2, wherein said construct is further free from association with transfection facilitating calcium phosphate.

6. The process of claim 2, wherein said DNA comprises a plasmid.

7. The process of claim 2, wherein said administration is intravenous or intranasal.

8. The process of claim 7, wherein said construct is injected.

9. The process of claim 7, wherein said administration is intravenous.

10. The process of claim 7, wherein said administration is intranasal.

11. The process of claim 2, wherein said immunogen is viral.

12. The process of claim 2, wherein said uptake is into tissue cells.

13. A process for inducing an effective immune response in a mammal to a pathogen comprising:

administering in vivo into a tissue of the mammal a construct comprising a noninfectious, nonintegrating DNA sequence encoding an immunogen, wherein said DNA is operably linked to a promoter, in an amount sufficient that uptake of said construct into cells of the mammal occurs, and sufficient expression results, to induce the effective immune response to a pathogen,

wherein said construct is free from association with transfection-facilitating proteins, viral particles, liposomal formulations and charged lipids.

14. The process of claim 13, wherein said construct is introduced into mucous membrane tissue.

15. The process of claim 13, wherein the mammal is human.

16. The process of claim 13, wherein said construct is further free from association with transfection facilitating calcium phosphate.

17. The process of claim 13, wherein said DNA comprises a plasmid.

18. The process of claim 13, wherein said administration is intravenous or intranasal.

19. The process of claim 18, wherein said construct is injected.

20. The process of claim 18, wherein said administration is intravenous.

21. The process of claim 18, wherein said administration is intranasal.

22. The process of claim 13, wherein said immunogen is viral.

23. The process of claim 13, wherein said uptake is into tissue cells.

24. A process for generating a detectable antibody response in a vertebrate, comprising:

administering in vivo into a tissue of the vertebrate a construct comprising a noninfectious, nonintegrating DNA sequence encoding an immunogen, wherein said DNA is operably linked to a promoter, in an amount sufficient that uptake of said construct into cells of the vertebrate occurs, and sufficient expression results, to generate the detectable antibody response,

wherein said construct is free from association with transfection-facilitating proteins, viral particles, liposomal formulations and charged lipids.

25. The process of claim 24, wherein said construct is introduced into mucous membrane tissue.

26. The process of claim 24, wherein said construct is introduced into muscle.

27. The process of claim 24, wherein said construct is introduced into skin.

28. The process of claim 24, wherein said construct is further free from association with transfection facilitating calcium phosphate.

29. The process of claim 24, wherein said DNA comprises a plasmid.

30. The process of claim 24, wherein said administration is intramuscular, intravenous, intradermal subcutaneous, or intranasal.

31. The process of claim 30, wherein said construct is injected.

32. The process of claim 30, wherein said administration is intramuscular.

33. The process of claim 30, wherein said administration is intravenous.

34. The process of claim 30, wherein said administration is intradermal.

35. The process of claim 30, wherein said administration is subcutaneous.

36. The process of claim 30, wherein said administration is intranasal.

37. The process of claim 24, wherein said immunogen is viral.

38. The process of claim 24, wherein said uptake is into tissue cells.

39. The process of claim 24, wherein the vertebrate is a bird.

40. The process of claim 24, wherein the vertebrate is a fish.

41. A process for generating a detectable antibody response in a mammal, comprising:

injecting into muscle of the mammal a construct comprising a noninfectious, nonintegrating mRNA sequence encoding an immunogen in an amount sufficient that uptake of said construct occurs, and sufficient expression results, to generate the detectable antibody response,

wherein said construct is free from association with transfection-facilitating proteins, viral particles, liposomal formulations and charged lipids.

42. The process of claim 41, wherein the mamnal is human.

43. The process of claim 41, wherein said construct is further free from association with transfection including calcium phosphate.

44. The process of claim 41, wherein said immunogen is viral.

45. The process of claim 41, wherein said uptake is into said muscle.

46. A method of generating an effective immune response against a pathogen in a mammal, comprising:

administering in vivo into a tissue of a mammal in need of said immune response a composition comprising a DNA plasmid which directs synthesis of an immunogenic peptide or polypeptide in mammalian cells through association with a promoter, and a pharmaceutically acceptable carrier;

wherein said DNA plasmid is free from association with liposomal formulations and charged lipids;

wherein said DNA plasmid incorporates into the cells of said mammal; and

wherein sufficient expression of said immunogenic peptide or polypeptide occurs to generate said effective immune response against said pathogen in said mammal.

47. The method of claim 46, wherein said immunogenic peptide or polypeptide metabolically incorporates into the cell membranes of said mammal by binding to a class I major histocompatibility molecule, a class II major histocompatibility molecule, or a combination of class I and class II major histocompatibility molecules.

48. The method of claim 46, wherein said promoter is selected from the group consisting of a Rous sarcoma virus long terminal repeat (RSVLTR), a myeloproliferative sarcoma virus long terminal repeat (MPSVLTR), a simian virus 40 immediate early promoter (SV40IEP), a metallothionein promoter, and a human cytomegalovirus immediate early promoter (CMVIEP).

49. The method of claim 48, wherein said promoter is a human cytomegalovirus immediate early IEP promoter.

50. The method of claim 46, wherein said tissue is selected from the group consisting of muscle, skin, and blood.

51. The method of claim 46, wherein said composition is injected.

52. The method of claim 46, wherein said tissue is muscle.

53. The method of claim 46, wherein said immunogenic peptide or polypeptide is a viral protein.

54. A method of generating an effective immune response against a pathogen in a mammal, comprising:

administering in vivo into a tissue of a mammal in need of said immune response a composition consisting essentially of a DNA plasmid operably encoding an immunogenic peptide or polypeptide through association with a promoter, and a pharmaceutically acceptable carrier;

wherein said plasmid incorporates into the cells of said mammal; and

wherein sufficient expression of said immunogenic peptide or polypeptide occurs to generate said effective immune response against said pathogen in said mammal.

55. The method of claim 54, wherein said immunogenic peptide or polypeptide metabolically incorporates into the cell membranes of said mammal by binding to a class I major histocompatibility molecule, a class II major histocompatibility molecule, or a combination of class I and class II major histocompatibility molecules.

56. The method of claim 54, wherein said promoter is selected from the group consisting of a Rous sarcoma virus long terminal repeat (RSVLTR), a myeloproliferative sarcoma virus long terminal repeat (MPSVLTR), a simian virus 40 immediate early promoter (SV40IEP), a metallothionein promoter, and a human cytomegalovirus immediate early promoter (CMVIEP).

57. The method of claim 56, wherein said promoter is a human cytomegalovirus immediate early promoter.

58. The method of claim 54, wherein said tissue is selected from the group consisting of muscle, skin, and blood.

59. The method of claim 54, wherein said composition is injected.

60. The method of claim 54, wherein said tissue is muscle.

61. The method of claim 54, wherein said immunogenic peptide or polypeptide is a viral protein.

62. A method of generating an effective immune response against a pathogen in a mammal, comprising:

administering in vivo into a tissue of a mammal in need of said immune response a composition consisting of a DNA plasmid operably encoding an immunogenic peptide or polypeptide through association with a promoter, and a pharmaceutically acceptable carrier;

wherein said plasmid incorporates into the cells of said mammal; and

wherein sufficient expression of said immunogenic peptide or polypeptide occurs to generate said effective immune response against said pathogen in said mammal.

63. The method of claim 62, wherein said immunogenic peptide or polypeptide metabolically incorporates into the cell membranes of said mammal by binding to a class I major histocompatibility molecule, a class II major histocompatibility molecule, or a combination of class I and class II major histocompatibility molecules.

64. The method of claim 62, wherein said promoter is selected from the group consisting of a Rous sarcoma virus long terminal repeat (RSVLTR), a myeloproliferative sarcoma virus long terminal repeat (MPSVLTR), a simian virus 40 immediate early promoter (SV40IEP), a metallothionein promoter, and a human cytomegalovirus immediate early promoter (CMVIEP).

65. The method of claim 64, wherein said promoter is a human cytomegalovirus immediate early promoter.

66. The method of claim 62, wherein said tissue is selected from the group consisting of muscle, skin, and blood.

67. The method of claim 62, wherein said composition is injected.

68. The method of claim 62, wherein said tissue is muscle.

69. The method of claim 62, wherein said immunogenic peptide or polypeptide is a viral protein.

70. A method for immunizing a mammal against infection by a pathogen, comprising:

obtaining an injectable preparation consisting of a pharmaceutically acceptable injectable carrier containing an expressible polynucleotide coding for an immunogenic peptide or polypeptide, wherein said polynucleotide is a DNA plasmid operably encoding said immunogenic peptide or polypeptide through association with a promoter;

introducing said injectable preparation into a mammal in need of said immunization, whereby said polynucleotide incorporates into a cell where an immunogenic translation product of said polynucleotide is formed, and said product is processed and presented by said cell in the context of the major histocompatibility complex, thereby eliciting an effective immune response against said pathogen.

71. A method of generating an effective immune response against a pathogen in a mammal, comprising:

administering in vivo into muscle tissue of a mammal in need of said immune response a composition comprising a polynucleotide which directs synthesis of an immunogenic peptide or polypeptide in mammalian cells, and a pharmaceutically acceptable carrier;

wherein said polynucleotide is selected from the group consisting of a messenger RNA and a DNA plasmid operably encoding said immunogenic peptide or polypeptide through association with a promoter;

wherein said polynucleotide is free from association with liposomal formulations and charged lipids;

wherein said polynucleotide incorporates into the cells of said mammal; and

wherein sufficient expression of said immunogenic peptide or polypeptide occurs to generate said effective immune response against said pathogen in said mammal.

72. The method of claim 71, wherein said polynucleotide is messenger RNA.

73. The method of claim 72, wherein said immunogenic peptide or polypeptide metabolically incorporates into the cell membranes of said mammal by binding to a class I major histocompatibility molecule, a class II major histocompatibility molecule, or a combination of class I and class II major histocompatibility molecules.

74. The method of claim 72, wherein said composition is injected.

75. The method of claim 72, wherein said immunogenic peptide or polypeptide is a viral protein.

76. A process for generating a detectable antibody response in a mammal, comprising:

administering in vivo into a tissue of the mammal a construct comprising a noninfectious, nonintegrating DNA sequence encoding an immunogen, wherein said DNA is operably linked to a promoter, in an amount sufficient that uptake of said construct into cells of the mammal occurs, and sufficient expression results, to generate the detectable antibody response,

wherein said construct is free from association with any transfection-facilitating material.

77. A process for inducing an effective immune response in a mammal to a pathogen comprising:

administering in vivo into a tissue of the mammal a construct comprising a noninfectious, nonintegrating DNA sequence encoding an immunogen, wherein said DNA is operably linked to a promoter, in an amount sufficient that uptake of said construct into cells of the mammal occurs, and sufficient expression results, to induce the effective immune response to a pathogen,

wherein said construct is free from association with any transfection-facilitating material.

78. A process for generating a detectable antibody response in a vertebrate, comprising:

administering in vivo into a tissue of the vertebrate a construct comprising a noninfectious, nonintegrating DNA sequence encoding an immunogen, wherein said DNA is operably linked to a promoter, in an amount sufficient that uptake of said construct into cells of the vertebrate occurs, and sufficient expression results, to generate the detectable antibody response,

wherein said construct is free from association with any transfection-facilitating material.

79. A process for generating a detectable antibody response in a mammal, comprising:

injecting into muscle of the mammal a construct comprising a noninfectious, nonintegrating mRNA sequence encoding an immunogen in an amount sufficient that uptake of said construct occurs, and sufficient expression results, to generate the detectable antibody response,

wherein said construct is free from association with transfection-facilitating material.