Methods for regualting hematopoiesis using CpG-oligonucleotides

• US 20040234512A1
• Filed: 06/25/2004
• Published: 11/25/2004
• Est. Priority Date: 05/14/1998
• Status: Abandoned Application

• Current Assignee: Coley Pharmaceutical GmbH (Pfizer Inc.)
• Original Assignee: Coley Pharmaceutical GmbH (Pfizer Inc.)
• Inventors: Wagner, Hermann, Lipford, Grayson
• Application Number:
  US10/877,369
• Publication Number:
  US 20040234512A1
• Time in Patent Office: Days
• US Class Current: 424/93.21
• CPC Class Codes: A61K 2039/55555 Liposomes; Vesicles, e.g. nanoparticles; Spheres, e.g. nanospheres; Polymers
• CPC Class Codes: A61K 2039/55561 CpG containing adjuvants; Oligonucleotide containing adjuvants
• CPC Class Codes: A61K 2039/55566 Emulsions, e.g. Freund's adjuvant, MF59
• CPC Class Codes: A61K 31/70 Carbohydrates; Sugars; Derivatives thereof sorbitol A61K31/047
• CPC Class Codes: A61K 39/00 Medicinal preparations containing antigens or antibodies materials for immunoassay G01N33/53
• CPC Class Codes: A61K 39/39 characterised by the immunostimulating additives, e.g. chemical adjuvants
• CPC Class Codes: C12N 15/117 Nucleic acids having immunomodulatory properties, e.g. containing CpG-motifs
• CPC Class Codes: C12N 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/345 having at least two different backbone modifications
• 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

The invention relates to methods for regulating hematopoiesis using CpG containing oligonucleotides. In particular, the invention relates to methods of treating thrombopoiesis and anemia by regulating hematopoiesis. The invention also relates to methods of regulating immune system remodeling by administering CpG oligonucleotides to control hematopoiesis.

Claims

1-65. -65. (canceled).

66. A method for enhancing an antigen-specific immune response to an antigen comprising administering a CpG oligonucleotide to a subject at least 48 hours prior to exposure of the subject to the antigen.

67. The method of claim 66, wherein the CpG oligonucleotide is administered between about 48 hours and 40 days prior to exposure of the subject to the antigen.

68. The method of claim 66, wherein the immune response is a Th1 response.

69. The method of claim 66, wherein the immune response includes production of an antibody.

70. The method of claim 69, wherein the antibody is of the IgG2 class.

71. The method of claim 66, wherein the immune response includes release of interferon-gamma.

72. The method of claim 66, wherein the immune response includes a cytotoxic T lymphocyte (CTL) response.

73. The method of claim 66, wherein the antigen is an antigen or intact microorganism associated with an infectious disease.

74. The method of claim 73, wherein the infectious disease is selected from the group consisting of viral, bacterial, mycobacterial, and parasitic disease.

75. The method of claim 66, wherein the CpG oligonucleotide is a DNA or RNA oligonucleotide comprising an unmethylated cytosine-guanine dinucleotide sequence.

76. The method of claim 66, wherein the CpG oligonucleotide is 8-100 nucleotides long and comprises a formula 5′

X₁CGX₂3′

, wherein C and G are unmethylated and X₁ and X₂ are nucleotides.

77. The method of claim 66, wherein the CpG oligonucleotide is 8-100 nucleotides long and comprises a formula 5′

X₁X₂CGX₃X₄3′

, wherein X₁X₂ are nucleotides selected from GpT, GpG, GpA, and ApA, and wherein X₃X₄ are nucleotides selected from TpT, CpT, and GpT.

78. The method of claim 66, wherein the CpG oligonucleotide is 8-100 nucleotides long and comprises a formula 5′

TCNX₁X₂CGX₃X₄3′

, wherein X₁, X₂, X₃, and X₄ are nucleotides and N is a nucleic acid sequence composed of from about 0-25 nucleotides.

79. The method of claim 66, wherein the CpG oligonucleotide is 8-100 nucleotides long and comprises a formula 5′

TCNX₁X₂CGX₃X₄3′

, wherein N is a nucleic acid sequence composed of from about 0-25 nucleotides, X₁X₂ are nucleotides selected from GpT, GpG, GpA, and ApA, and wherein X₃X₄ are nucleotides selected from TpT, CpT, and GpT.

80. The method of claim 66, wherein the CpG oligonucleotide is 8-100 nucleotides long and comprises a formula 5′

N₁X₁CGX₂N₂3′

, wherein at least one nucleotide separates consecutive CpGs;

N₁ is adenine, guanine, or thymine;

X₂ is cytosine, adenine, or thymine;

N is any nucleotide; and

N₁ and N₂ are nucleic acid sequences composed of from about 0-25 N'"'"'s.

81. The method of claim 66, wherein the CpG oligonucleotide is 8-100 nucleotides long and comprises a formula 5′

N₁X₁X₂CGX₃X₄N₂3′

, wherein at least one nucleotide separates consecutive CpGs;

X₁X₂ is selected from the group consisting of TpT, CpT, TpC, and ApT;

X₃X₄ is selected from the group consisting of GpT, GpA, ApA and ApT;

N is any nucleotide and N₁ and N₂ are nucleic acid sequences composed of from about 0-25 N'"'"'s.

82. The method of claim 66, wherein the CpG oligonucleotide comprises a phosphorothioate or phosphorodithioate backbone modification.

83. The method of claim 66, wherein the CpG oligonucleotide comprises a sequence selected from a group consisting of AACGTT, AGCGTT, GACGTT, GGCGTT, GTCGTT, GTCGCT, GGCGCT, GACGCT, and AACGCT.

84. The method of claim 66, wherein the CpG oligonucleotide is administered between about 2 days and at least 28 days prior to exposure of the subject to the antigen.

85. The method of claim 66, wherein the CpG oligonucleotide is administered between about 3 days and about 8 days prior to exposure of the subject to the antigen.

86. The method of claim 66, wherein the CpG oligonucleotide is administered via a mucosal or systemic route.

87. The method of claim 86, wherein the mucosal route is intranasal or intratracheal.

88. The method of claim 86, wherein the systemic route is subcutaneous or intravenous.

89. A method of immunizing a subject against an antigen comprising administering to the subject a CpG oligonucleotide at least 48 hours prior to exposing the subject to the antigen.

90. The method of claim 89, wherein the CpG oligonucleotide is administered between about 48 hours and 40 days prior to exposing the subject to the antigen.

91. The method of claim 89, wherein the CpG oligonucleotide is a DNA or RNA oligonucleotide comprising an unmethylated cytosine-guanine dinucleotide sequence.

92. The method of claim 89, wherein the CpG oligonucleotide is 8-100 nucleotides long and comprises a formula 5′

X₁CGX₂3′

, wherein C and G are unmethylated and X₁ and X₂ are nucleotides.

93. The method of claim 89, wherein the CpG oligonucleotide is 8-100 nucleotides long and comprises a formula 5′

X₁X₂CGX₃X₄3′

, wherein X₁X₂ are nucleotides selected from GpT, GpG, GpA, and ApA, and wherein X₃X₄ are nucleotides selected from TpT, CpT, and GpT.

94. The method of claim 89, wherein the CpG oligonucleotide is 8-100 nucleotides long and comprises a formula 5′

TCNX₁X₂CGX₃X₄3′

, wherein X₁, X₂, X₃, and X₄ are nucleotides and N is a nucleic acid sequence composed of from about 0-25 nucleotides.

95. The method of claim 89, wherein the CpG oligonucleotide is 8-100 nucleotides long and comprises a formula 5′

TCNX₁X₂CGX₃X₄3′

, wherein N is a nucleic acid sequence composed of from about 0-25 nucleotides, X₁X₂ are nucleotides selected from GpT, GpG, GpA, and ApA, and wherein X₃X₄ are nucleotides selected from TpT, CpT, and GpT.

96. The method of claim 89, wherein the CpG oligonucleotide is 8-100 nucleotides long and comprises a formula 5′

N₁X₁CGX₂N₂3′

, wherein at least one nucleotide separates consecutive CpGs;

N₁ is adenine, guanine, or thymine;

X₂ is cytosine, adenine, or thymine;

N is any nucleotide; and

N₁ and N₂ are nucleic acid sequences composed of from about 0-25 N'"'"'s.

97. The method of claim 89, wherein the CpG oligonucleotide is 8-100 nucleotides long and comprises a formula 5′

N₁X₁X₂CGX₃X₄N₂3′

, wherein at least one nucleotide separates consecutive CpGs;

X₁X₂ is selected from the group consisting of TpT, CpT, TpC, and ApT;

X₃X₄ is selected from the group consisting of GpT, GpA, ApA and ApT;

N is any nucleotide and N₁ and N₂ are nucleic acid sequences composed of from about 0-25 N'"'"'s.

98. The method of claim 89, wherein the CpG oligonucleotide comprises a phosphorothioate or phosphorodithioate backbone modification.

99. The method of claim 89, wherein the CpG oligonucleotide comprises a sequence selected from a group consisting of AACGTT, AGCGTT, GACGTT, GGCGTT, GTCGTT, GTCGCT, GGCGCT, GACGCT, and AACGCT.

100. The method of claim 89, wherein the CpG oligonucleotide is administered between about 2 days and at least 28 days prior to exposing the subject to the antigen.

101. The method of claim 89, wherein the CpG oligonucleotide is administered between about 3 days and about 8 days prior to exposing the subject to the antigen.

102. The method of claim 89, wherein the antigen is an antigen or intact microorganism associated with an infectious disease.

103. The method of claim 102, wherein the infectious disease is selected from the group consisting of viral, bacterial, mycobacterial, and parasitic disease.

104. The method of claim 89, wherein the CpG oligonucleotide is administered via a mucosal or systemic route.

105. The method of claim 104, wherein the mucosal route is intranasal or intratracheal.

106. The method of claim 104, wherein the systemic route is subcutaneous or intravenous.

107. A method of inducing IgG2 antibody production comprising administering to a subject a CpG oligonucleotide at least 48 hours prior to administration of an antigen to the subject.

108. The method of claim 107, wherein the CpG oligonucleotide is administered between about 48 hours and 40 days prior to administration of the antigen.

109. The method of claim 107, wherein the CpG oligonucleotide is a DNA or RNA oligonucleotide comprising an unmethylated cytosine-guanine dinucleotide sequence.

110. The method of claim 107, wherein the CpG oligonucleotide is 8-100 nucleotides long and comprises a formula 5′

X₁CGX₂3′

, wherein C and G are unmethylated and X₁ and X₂ are nucleotides.

111. The method of claim 107, wherein the CpG oligonucleotide is 8-100 nucleotides long and comprises a formula 5′

X₁X₂CGX₃X₄3′

, wherein X₁X₂ are nucleotides selected from GpT, GpG, GpA, and ApA, and wherein X₃X₄ are nucleotides selected from TpT, CpT, and GpT.

112. The method of claim 107, wherein the CpG oligonucleotide is 8-100 nucleotides long and comprises a formula 5′

TCNX₁X₂CGX₃X₄3′

, wherein X₁, X₂, X₃, and X₄ are nucleotides and N is a nucleic acid sequence composed of from about 0-25 nucleotides.

113. The method of claim 107, wherein the CpG oligonucleotide is 8-100 nucleotides long and comprises a formula 5′

TCNX₁X₂CGX₃X₄3′

, wherein N is a nucleic acid sequence composed of from about 0-25 nucleotides, X₁X₂ are nucleotides selected from GpT, GpG, GpA, and ApA, and wherein X₃X₄ are nucleotides selected from TpT, CpT, and GpT.

114. The method of claim 107, wherein the CpG oligonucleotide is 8-100 nucleotides long and comprises a formula 5′

N₁X₁CGX₂N₂3′

, wherein at least one nucleotide separates consecutive CpGs;

N₁ is adenine, guanine, or thymine;

X₂ is cytosine, adenine, or thymine;

N is any nucleotide; and

N₁ and N₂ are nucleic acid sequences composed of from about 0-25 N'"'"'s.

115. The method of claim 107, wherein the CpG oligonucleotide is 8-100 nucleotides long and comprises a formula 5′

N₁X₁X₂CGX₃X₄N₂3′

, wherein at least one nucleotide separates consecutive CpGs;

X₁X₂ is selected from the group consisting of TpT, CpT, TpC, and ApT;

X₃X₄ is selected from the group consisting of GpT, GpA, ApA and ApT;

N is any nucleotide and N₁ and N₂ are nucleic acid sequences composed of from about 0-25 N'"'"'s.

116. The method of claim 107, wherein the CpG oligonucleotide comprises a phosphorothioate or phosphorodithioate backbone modification.

117. The method of claim 107, wherein the CpG oligonucleotide comprises a sequence selected from a group consisting of AACGTT, AGCGTT, GACGTT, GGCGTT, GTCGTT, GTCGCT, GGCGCT, GACGCT, and AACGCT.

118. The method of claim 107, wherein the CpG oligonucleotide is administered between about 2 days and at least 28 days prior to administration of the antigen.

119. The method of claim 107, wherein the CpG oligonucleotide is administered between about 3 days and about 8 days prior to administration of the antigen.

120. The method of claim 107, wherein the antigen is an antigen or intact microorganism associated with an infectious disease.

121. The method of claim 120, wherein the infectious disease is selected from the group consisting of viral, bacterial, mycobacterial, and parasitic disease.

122. The method of claim 107, wherein the CpG oligonucleotide is administered via a mucosal or systemic route.

123. The method of claim 122, wherein the mucosal route is intranasal or intratracheal.

124. The method of claim 122, wherein the systemic route is subcutaneous or intravenous.