Recombinant non-replicating virus expressing gm-csf and uses thereof to enhance immune responses

• US 20040091995A1
• Filed: 07/16/2003
• Published: 05/13/2004
• Est. Priority Date: 06/15/2001
• Status: Abandoned Application

• Current Assignee: Government Of The United States Of America, Represented By The Secretary, Department Of Health And Human Services, United States As Represented By The Secretary Department of Health and Human Services
• Original Assignee: Government Of The United States Of America, Represented By The Secretary, Department Of Health And Human Services
• Inventors: Kass, Erik, Schlom, Jeffrey, Greiner, John W., Panicali, Dennis
• Application Number:
  US10/297,168
• Publication Number:
  US 20040091995A1
• Time in Patent Office: Days
• US Class Current: 435/235.1
• CPC Class Codes: A61K 2039/525 Virus
• CPC Class Codes: A61K 2039/5256 expressing foreign proteins
• CPC Class Codes: A61K 2039/55522 Cytokines; Lymphokines; Interferons
• CPC Class Codes: A61K 38/193 Colony stimulating factors [CSF]
• CPC Class Codes: A61K 39/00 Medicinal preparations containing antigens or antibodies materials for immunoassay G01N33/53
• CPC Class Codes: A61K 39/0011 Cancer antigens
• CPC Class Codes: A61K 39/001106 Her-2/neu/ErbB2, Her-3/ErbB3 or Her 4/ErbB4
• CPC Class Codes: A61K 39/001151 p53
• CPC Class Codes: A61K 39/001156 Tyrosinase and tyrosinase related proteinases [TRP-1 or TRP-2]
• CPC Class Codes: A61K 39/001164 GTPases, e.g. Ras or Rho
• CPC Class Codes: A61K 39/00117 Mucins, e.g. MUC-1
• CPC Class Codes: A61K 39/001182 Carcinoembryonic antigen [CEA]
• CPC Class Codes: A61K 39/001186 MAGE
• CPC Class Codes: A61K 39/001188 NY-ESO
• CPC Class Codes: A61K 39/001191 Melan-A/MART
• CPC Class Codes: A61K 39/001192 Glycoprotein 100 [Gp100]
• CPC Class Codes: A61K 39/001194 Prostate specific antigen [PSA]
• CPC Class Codes: A61K 39/001195 Prostate specific membrane antigen [PSMA]
• CPC Class Codes: A61K 39/12 Viral antigens
• CPC Class Codes: A61K 39/21 Retroviridae, e.g. equine infectious anemia virus
• CPC Class Codes: A61K 39/39 : characterised by the immunostimulating additives, e.g. chemical adjuvants
• CPC Class Codes: A61K 39/4611 : T-cells, e.g. tumor infiltrating lymphocytes [TIL], lymphokine-activated killer cells [LAK] or regulatory T cells [Treg]
• CPC Class Codes: A61K 39/464482 : Carcinoembryonic antigen [CEA]
• 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 48/005 : characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered
• CPC Class Codes: C12N 15/86 : Viral vectors
• CPC Class Codes: C12N 2710/24043 : viral genome or elements thereof as genetic vector
• CPC Class Codes: C12N 2710/24143 : viral genome or elements thereof as genetic vector
• CPC Class Codes: C12N 2740/15022 : New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
• CPC Class Codes: C12N 2740/15034 : Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
• CPC Class Codes: C12N 2740/16022 : New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
• CPC Class Codes: C12N 2740/16034 : Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
• CPC Class Codes: C12N 7/00 : Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00
• CPC Class Codes: Y02A 50/30 : Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

Replication-defective recombinant poxvirus encoding granulocyte-macrophage colony-stimulating factor (GM-CSF) are disclosed for use in enriching an immunization site with antigen-presenting cells (APC), for enhancing an immunological response to antigen or immunological epitopes by functioning as a biological adjuvant, for prevention or treatment of neutropenia, and for the treatment of myeloidysplastic syndromes. Compositions comprising a replication-defective recombinant virus encoding GM-CSF alone or in combination with a recombinant virus encoding an antigen and optionally encoding an immunostimulatory molecule are disclosed for enhancing antigen-specific immunological responses, in particular enhancing tumor antigen responses for anti-tumor therapy. Methods for enriching an immunization site with APC and for enhancing immunological responses to an antigen or immunological epitope using replication-defective recombinant poxvirus encoding GM-CSF are disclosed. The superiority of the use of a replication-defective recombinant avian poxvirus encoding GM-CSF over the use of recombinant GM-CSF is described.

Claims

1. A pharmaceutical composition comprising a replication-defective virus encoding granulocyte-monocyte-colony stimulating factor (GM-CSF) and a pharmaceutically acceptable carrier.

2. The pharmaceutical composition according to claim 1, wherein the replication-defective virus encodes human GM-CSF.

3. The pharmaceutical composition according to claim 1, wherein the virus is produced using a plasmid vector designated pT5052 deposited with the American Type Culture Collection under Accession No. PTA-2099.

4. The pharmaceutical composition according to claim 1, wherein the replication-defective virus is a poxvirus.

5. The pharmaceutical composition according to claim 1 wherein the replication-defective virus is an avipox virus.

6. The pharmaceutical composition according to claim 5 wherein the avipox virus is selected from the group consisting of fowlpox virus, canarypox virus, MVA, and derivatives thereof.

7. The pharmaceutical composition according to any one of claims 1-6, wherein the replication-defective virus further encodes at least one antigen or immunological epitope thereof.

8. The pharmaceutical composition according to claim 7, wherein the antigen is selected from the group consisting of tumor specific antigen, tumor associated antigen, tissue-specific antigen, bacterial antigen, viral antigen, yeast antigen, fungal antigen, protozoan antigen, parasite antigen and mitogen.

9. The pharmaceutical composition according to claim 7, wherein the immunological epitope comprises at least one amino acid sequence selected from the group consisting of SEQ ID NO:

1 through SEQ ID NO;

36 and combinations thereof.

10. The pharmaceutical composition according to claim 1 further comprising a vector encoding at least one costimulatory molecule.

11. The pharmaceutical composition according to claim 10, wherein the costimulatory molecule is B7.1.

12. The pharmaceutical composition according to claim 10, wherein the costimulatory molecule is B7.1/ULA-3/ICAM-1.

13. The pharmaceutical composition according to any one of claims 1-12, further comprising a vector encoding alpha interferon, beta interferon, or gamma interferon.

14. The pharmaceutical composition according to any one of claims 1-12 further comprising a vector encoding at least one cytokine.

15. The pharmaceutical composition according to claim 14, wherein the cytokine is IL-12.

16. The pharmaceutical composition according to any one of claims 1-6, further comprising an Fc receptor-directed bispecific antibody.

17. The pharmaceutical composition according to claim 16, wherein the Fc receptor is selected from the group consisting of Fcγ

RI (CD64), Fcγ

RII (CD32) and Fcγ

RIII (CD16).

18. The pharmaceutical composition according to claim 16, wherein the antibody is an anti-CD3-directed antibody.

19. The pharmaceutical composition according to any one of claims 16-18, wherein the bispecific antibody has tumor-directed specificity.

20. The pharmaceutical composition according to claim 19, wherein the specificity is selected from the group consisting of HER-2/neu, EGF-receptor, CD15 antigen and a EpCAM molecule.

21. The pharmaceutical composition according to claims 1-16, further comprising at least one antigen or immunological epitope source, and optionally a conventional adjuvant.

22. The pharmaceutical composition according to claim 21, wherein the conventional adjuvant is selected from the group consisting of Ribi Detox™

, alum, QS-21, Freund'"'"'s complete adjuvant, and Freund'"'"'s incomplete adjuvant.

23. The pharmaceutical composition according to claim 21, wherein the antigen or immunological epitope source is a protein, peptide, antibody, anti-idiotypic antibody, lipid, carbohydrate, cell, cell extract, cell fragment, DNA encoding an antigen, or encoding an immunological epitope thereof RNA encoding an antigen or encoding an immunological epitope thereof, or a vector encoding at least one antigen or immunological epitope thereof.

24. The composition according to claim 21, wherein the antigen is selected from the group consisting of a tumor specific antigen, tumor associated antigen, tissue-specific antigen, bacterial antigen, viral antigen, yeast antigen, fungal antigen, protozoan antigen, parasite antigen and mitogen.

25. The composition according to claim 21, wherein the immunological epitope comprises at least one amino acid sequence selected from the group consisting of SEQ ID NO:

1 through SEQ ID NO;

36, and combinations thereof.

26. The composition according to claim 24, wherein the bacterial antigen is derived from a bacterium selected from the group consisting of Chlamydia, Mycobacteria, Legionella, Meningiococcus, Group A Streptococcus, Hemophilus influenzae, Salmonella, and Listeria

27. The composition according to claim 24, wherein the viral antigen is derived from a virus selected from the group consisting of Lentivirus, retrovirus, Herpes virus, Hepatitis virus, Orthomyxovirus and Papillomavirus.

28. The composition according to claim 27, wherein the Lentivirus is HIV-1 or HIV-2.

29. The composition according to claim 27, wherein the Herpes virus is HSV or CMV.

30. The composition according to claim 27, wherein the Hepatitis virus is Hepatitis A, Hepatitis B, Hepatitis C, Hepatitis D or Hepatitis E.

31. The composition according to claim 27, wherein the orthomyxovirus is influenza virus.

32. The composition according to claim 24, wherein the tumor associated antigen, tumor specific antigen or tissue-specific antigen is selected from the group consisting of CEA, MART-1, MAGE-1, MAGE-3, GP-100, MUC-1, MUC-2, point mutated ras oncogene, normal or point mutated p53, overexpressed p53, CA-125, PSA, PSMA, C-erb/132, BRCA I, BRCA II, PSMA, tyrosinase, TRP-1, TRP-2, NY-ESO-1, TAG72, KSA, HER-2/neu, bcr-abl, pax3-fkhr, ews-fli-1, modified TAAs, splice variants of TAAs, functional epitopes and epitope agonists thereof.

33. The composition according to claim 24, wherein the yeast or fungal antigen is derived from a yeast or fungus selected from the group consisting of Aspergillus, Nocardia, Histoplasmosis, Candida, and Cryptosporidia.

34. The composition according to claim 24, wherein the parasitic antigen is derived from a Plasmodium species, Toxoplasma gondii, Pneumocystis carinii, Trypasosoma species, or Leishmania species.

35. The composition according to claim 23, wherein the vector encoding an antigen or immunological epitope thereof is selected from the group consisting of poxvirus, adenovirus, Herpes virus, alphavirus, picomavirus, iridovirus, DNA plasmids, and RNA.

36. The composition according to claim 35, wherein the poxvirus is orthopox, avipox, capripox or suipox.

37. The composition according to claim 35, wherein the antigen is selected from the group consisting of a tumor specific antigen, tumor associated antigen, tissue-specific antigen, bacterial antigen, viral antigen, yeast antigen, fungal antigen, protozoan antigen, parasite antigen, and mitogen.

38. The composition according to claim 35, wherein the immunological epitope comprises at least one amino acid sequence selected from the group consisting of SEQ ID NO:

1 through SEQ ID NO;

36, and combinations thereof.

39. The composition according to claim 23, wherein the antigen or immunological epitope source is a vector encoding at least one antigen or antigenic epitope thereof.

40. The composition according to claim 23, wherein the vector encodes at least one costimulatory molecule selected from the group consisting of B7-1, B7-2, ICAM-1, LFA-3, 4-1BBL, CD59, CD40, CD70, OX-40L, VCAM-1, mammalian homologs thereof and combinations thereof.

41. The composition according to claim 40, wherein the costimulatory molecule is B7.1.

42. The compositon according to claim 40, wherein the costimulatory molecules are at least B7-1, ICAM-1 and LFA-3.

43. The composition according to claim 40, wherein the recombinant replication-defective virus encoding GM-CSF is avipox, the antigen source is a vector encoding CEA or epitope thereof, and the costimulatory molecule is B7 or B7.1/LFA-3/ICAM-1.

44. The composition according to any one of claims 1-15, further comprising a cytokine, chemokine or Flt-3L.

45. The composition according to any one of claims 1-15, further comprising at least one antibiotic, antifungal agent, anti-viral agent or combinations thereof.

46. The composition according to any one of claims 1-6, further comprising erythropoietin.

47. The composition according to claim 46, wherein the erythropoietin is recombinantly produced.

48. A host cell infected, transfected or induced with the recombinant replication-defective virus encoding GM-CSF according to any of claims 1-15 and 21-47.

49. The host cell according to claim 48, wherein the host cell is an antigen presenting cell or precursor thereof, a premalignant cell, a hyperplastic cell, tumor cell, or a tumor cell fused to an antigen presenting cell.

50. The host cell according to claim 49, wherein the antigen presenting cell is a dendritic cell or precursor or derivative thereof, a monocyte, macrophage, B-cell, fibroblast or muscle cell.

51. The host cell according to any one of claims 49 or 50, wherein the antigen presenting cell is derived from bone marrow, spleen, skin, peripheral blood, tumor, lymph node, or muscle.

52. The host cell according to claim 50, wherein the derivative is a TNF α

-treated dendritic cell, a CD40-treated dendritic cell, or a subpopulation of adherent cells.

64. A method of enhancing an immune response in an individual comprising administration of the composition according to any one of claims 1-15 and 21-47 in an amount sufficient to enhance the immune response.

65. The method according to claim 64, wherein a route of administration is intravenous, subcutaneous, intralymphatic, intratumoral, intradermal, intramuscular, intraperitoneal, intrarectal, intravaginal, intranasal, oral, via bladder instillation, intranasal, intraarterial, intravesical or via scarification.

66. The method according to claim 64, wherein the enhancement is migration of antigen presenting cells at an injection site, regional lymph node at a tumor site or combination thereof.

67. The method according to claim 66, wherein the antigen presenting cells express CD11c⁺/I-Ab⁺, MHC Class II, or combination thereof.

68. The method according to claim 66, wherein the enhancement is of antigen presenting cell proliferation, function or combination thereof.

69. The method according to claim 64, wherein the enhanced immune response is a cell mediated or humoral response.

70. The method according to claim 64, wherein the enhancement is of CD4⁺ T cell activation, CD8⁺ T cell activation, or combination thereof.

71. The method according to claim 64;

wherein the enhancement is in IL-2 production, IFN-γ

production, TNF-α

production, or combinations thereof.

76. A method of enhancing an anti-tumor response in an individual with a tumor comprising administration of the composition according to claims 1-15 or 21-47 in an amount effective to enhance the anti-tumor response.

77. The method according to claim 76, further comprising administration of a target antigen or immunological epitope thereof, cell expressing a target antigen or immunological epitope thereof, or cells pulsed with a target antigen or immunological epitope thereof.

78. The method according to any one of claim 76 or 77, wherein the composition is directly injected in situ into a tumor or adjacent to a tumor.

79. The method according to claim 78, wherein the tumor is a head tumor, neck tumor, melanoma, breast tumor, pancreatic tumor, prostate tumor, colorectal tumor, or metastatic tumor.

80. The method according to claim 79, wherein the tumor is a metastatic breast skin lesion.

81. The method according to any one of claims 76-79, wherein the composition is injected during surgery.

82. The method according to claim 81, wherein the tumor is a colonrectal cancer or pancreatic cancer.

83. The method according to any one of claims 76-79, wherein the composition is injected into a lymph node distal to or draining a tumor site.

84. The method according to any one of claims 76-79, further comprising the administration of activated, target antigen specific lymphocytes.

85. The method according to any one of claims 76-79, wherein the anti-tumor response is tumor regression, increase in disease-free interval, or increase in survival.

86. A method of enhancing an immune response in an individual comprising administration of a cell according to any of claims 48-62 in an amount effective to enhance an immune response.

87. A method of enhancing an immune response in an individual comprising administration of a tumor cell, or precursors thereof according to claim 49 or 54 in an amount effective to enhance an immune response.

88. The method according to any one of claims 86 or 87, wherein the cells are autologous, syngeneic or allogeneic with the individual.

89. The method according to any one of claims 86 or 87, wherein the cells have been pulsed with a target antigen or epitope thereof.

90. The method according to any one of claims 86 or 87, further comprising the administration of a target cell, target antigen or immunological epitope thereof.

91. The method according to any one of claims 86-90, further comprising the administration of activated, target antigen specific lymphocytes.

53. A dendritic cell or precursor thereof comprising a foreign nucleic acid sequence encoding GM-CSF provided by a recombinant replication-defective virus.

55. The cell according to claims 53 or 54, wherein the cell further comprises a foreign nucleic acid sequence encoding at least one costimulatory molecule.

56. The cell according to claim 55, wherein the costimulatory molecules are selected from the group consisting of B7-1, B7-2, ICAM-1, LFA-3, 4-1BBL, CD59, CD40, CD70, OX-40L, VCAM-1, mammalian homologs thereof and combinations thereof.

57. The cells according to claims 56, wherein the costimulatory molecule is B7.1.

58. The cells according to claim 56, wherein the multiple costimulatory molecules are at least B7-1, ICAM-1 and LFA-3.

59. The cells according to any one of claims 53-58 further comprising a foreign nucleic acid sequence encoding at least one target antigen or immunological epitope thereof.

60. The cells according to claim 59, wherein the foreign nucleic acid sequence encoding at least one target antigen or immunological epitope thereof is provided by a recombinant vector, RNA or DNA from a tumor cell lysate, or by fusion with a tumor cell comprising said sequence.

61. The cells according to any one of claims 59 or 60, wherein the target antigen or immunological epitope thereof is selected from the group consisting of a tumor specific antigen, tumor associated antigen, tissue-specific antigen, bacterial antigen, viral antigen, yeast antigen, fungal antigen, protozoan antigen, parasite antigen and mitogen.

62. The cells according to any one of claims 59 or 60, wherein the immunological epitope comprises at least one amino acid sequence selected from the group consisting of SEQ ID NO:

1 through SEQ ID NO;

36, and combinations thereof.

63. A pharmaceutical composition comprising the cells according to any one of claims 53-62, and optionally an exogenous source of target antigen or immunological epitope thereof.

54. A tumor cell or precursor thereof comprising a foreign nucleic acid sequence encoding GM-CSF provided by a recombinant replication-defective virus.

72. A method of enhancing an antigen-specific T-cell response in an individual to a target antigen or immunological epitope thereof comprising administering a recombinant replication-defective poxvirus encoding GM-CSF in combination with a recombinant virus comprising a nucleic acid sequence encoding a target antigen or immunological epitope thereof and optionally also comprising a foreign nucleic acid sequence encoding at least one B7 molecule, a foreign nucleic acid sequence encoding ICAM-1, and a nucleic acid sequence encoding LFA-3, in an amount effective to enhance at least one T-cell response.

73. The method according to claim 72, wherein the enhancement is of CD4⁺ T cell activation, CD8⁺ T cell activation, or combination thereof.

74. The method according to claim 72, wherein the enhancement is in IL2 production, IFN-γ

production or combination thereof.

75. The method according to claim 72, wherein the enhancement is of antigen-specific cytotoxicity.

92. A method of enhancing an immune response to an antigen or immunological epitope thereof in an individual comprising administration of a first recombinant vector encoding GM-CSF followed by administration of a second recombinant vector encoding GM-CSF wherein at least one recombinant vector is a replication-defective virus.

93. The method according to claim 92, wherein the replication defective virus is selected from the group consisting of poxvirus, herpes virus, adenovirus and adeno-associated virus.

94. The method according to claim 93, wherein the poxvirus is selected from the group consisting of fowlpox, canary pox and a Modified Vaccinia Ankara strain.

95. The method according to claim 92, wherein a second recombinant vector is replication competent.

96. The method according to claim 95, wherein the second vector is vaccinia.

97. A method of treating neutropenia in an individual comprising administration of a recombinant replication-defective virus encoding GM-CSF in an amount effective to treat the neutropenia.

98. The method according to claim 97, further comprising the administration of an antibiotic, antifungal agent, antiparasite agent, or antiviral agent.

99. The method according to claim 97, wherein the neutropenia is resultant from chemotherapy, corticosteroid therapy, irradiation, or an infection.

100. The method according to any one of claims 97-99, wherein a dose raises the neutrophil count to normal levels.

101. A method for treating cytopenias in patients with myeloidysplastic syndrome comprising administration of a recombinant replication-defective virus encoding GM-CSF in combination with erythropoietin in an amount effective to treat the cytopenia.

102. The method according to claim 101, wherein a dose in the range of about 10⁵ to about 10¹⁰ pfu of the recombinant replication-defective virus encoding GM-CSF is administered at a weekly or monthly interval.

103. The method according to any one of claims 101 or 102, wherein a dose in the range of about 150 to about 300 u/k of erythropoietin is administered.

104. The method according to any one of claims 101-104, wherein the dose is provided on alternate days.

105. The method according to any one of claims 101-104, wherein the treatment increases the neutrophil count and erythroid precursors.

106. A method of enhancing an antitumor immune response comprising administration of a replication-defective virus encoding GM-CSF in combination with a bispecific antibody.

107. The method according to claim 106, wherein the bispecific antibody has tumor-directed specificity.

108. The method according to claim 107, wherein the bispecific antibody has a specificity selected from the group consisting of HER-2/neu, EGF-receptor, CD15 antigen and a EpCAM molecule.

109. The method according to any one of claims 107-108, wherein the bispecific antibody is directed to a tumor cell epitope and a cytotoxin trigger molecule.

110. A method of enhancing an immune response to a vaccination comprising administration of a replication-defective virus encoding GM-CSF at the vaccination site or regional lymph node in an amount effective to enhance the immune response to a vaccine.

111. The method according to claim 110, wherein the vaccine comprises at least one tumor antigen or tumor associated antigen.

112. The method according to claim 110, wherein the vaccine is selected from the group consisting of DPT vaccine, Td vaccine, DtaP vaccine, Hib vaccine, DtaP-Hib vaccine, MMR vaccine, Hepatitis A vaccine, Hepatitis B vaccine, Lyme'"'"'s disease vaccine, influenza vaccine, tetravalent meningococcal polysaccharide, pneumococcal polysaccharide vaccine, anthrax vaccine, cholera vaccine, plague vaccine, yellow fever vaccine and Bacillus Calmette-Guerin vaccine.

113. An immunological adjuvant comprising a replication-defective virus encoding granulocyte-monocyte-colony stimulating factor.

114. A plasmid vector encoding human granulocyte-monocyte colony stimulating factor deposited with the American Type Culture Collection under Accession No. PTA-2099.