Circovirus sequences associated with piglet weight loss disease (PWD)

• US 20070253971A1
• Filed: 07/17/2006
• Published: 11/01/2007
• Est. Priority Date: 12/05/1997
• Status: Active Grant

• Current Assignee: Zoetis Services LLC (Zoetis Incorporated)
• Original Assignee: Wyeth (Pfizer Inc.)
• Inventors: Hutet, Evelyne, Blanchard, Philippe, Cariolet, Roland, Mahe, Dominique, Arnauld, Claire, Le Cann, Pierre, Truong, Catherine, Albina, Emmanuel, Madec, Francois, Jestin, Andre
• Granted Patent: US 7,407,803 B2
• Time in Patent Office: Days
• US Class Current: 424/186.100
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: A61K 2039/525 Virus
• CPC Class Codes: A61K 2039/5252 inactivated (killed)
• CPC Class Codes: A61K 2039/5254 avirulent or attenuated
• CPC Class Codes: A61K 2039/5256 expressing foreign proteins
• CPC Class Codes: A61K 2039/53 DNA (RNA) vaccination
• CPC Class Codes: A61K 2039/55 characterised by the host/recipient, e.g. newborn with maternal antibodies
• CPC Class Codes: A61K 2039/552 Veterinary vaccine
• CPC Class Codes: A61K 2039/55522 Cytokines; Lymphokines; Interferons
• CPC Class Codes: A61K 2039/55566 Emulsions, e.g. Freund's adjuvant, MF59
• CPC Class Codes: A61K 2039/58 raising an immune response against a target which is not the antigen used for immunisation
• CPC Class Codes: A61K 39/00 Medicinal preparations containing antigens or antibodies materials for immunoassay G01N33/53
• CPC Class Codes: A61K 39/12 Viral antigens
• 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: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 31/16 for influenza or rhinoviruses
• CPC Class Codes: A61P 31/20 for DNA viruses
• CPC Class Codes: A61P 31/22 for herpes viruses
• CPC Class Codes: A61P 33/00 Antiparasitic agents
• CPC Class Codes: A61P 37/02 : Immunomodulators
• CPC Class Codes: C07K 14/005 : from viruses
• CPC Class Codes: C07K 2319/55 : containing a fusion with a toxin, e.g. diphteria toxin
• CPC Class Codes: C12N 2710/14143 : viral genome or elements thereof as genetic vector
• CPC Class Codes: C12N 2750/10021 : Viruses as such, e.g. new isolates, mutants or their genomic sequences
• CPC Class Codes: C12N 2750/10022 : New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
• CPC Class Codes: C12N 2750/10034 : Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
• CPC Class Codes: C12N 2750/10051 : Methods of production or purification of viral material
• CPC Class Codes: C12N 2750/10061 : Methods of inactivation or attenuation
• 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: G01N 2333/01 : DNA viruses
• CPC Class Codes: G01N 2469/20 : Detection of antibodies in sample from host which are directed against antigens from microorganisms
• CPC Class Codes: G01N 33/56983 : Viruses
• CPC Class Codes: Y10T 428/13 : Hollow or container type article [e.g., tube, vase, etc.]

Abstract

The genome sequences and the nucleotide sequences coding for the PWD circovirus polypeptides, such as the circovirus structural and non-strucutral polypeptides, vectors including the sequences, and cells and animals transformed by the vectors are provided. Methods for detecting the nucleic acids or polypeptides, and kits for diagnosing infection by a PWD circovirus, also are provided. Method for selecting compounds capable of modulating the viral infection are further provided. Pharmaceutical, including vaccines, compositions for preventing and/or treating viral infections caused by PWD circovirus and the use of vectors for preventing and/or treating diseases also are provided.

Claims

1-51. -51. (canceled)

52. An immunogenic composition comprising a vector that contains and expresses in vivo in a porcine host cell, a nucleic acid molecule or fragment thereof encoding a porcine circovirus type B antigen and a pharmaceutically acceptable vehicle or veterinary vehicle.

53. The immunogenic composition of claim 52, wherein the vector is a viral vector.

54. The immunogenic composition of claim 52, wherein the vector is a plasmid.

55. The immunogenic composition of claim 52, wherein the composition induces an immune response in a porcine host against type B porcine circovirus.

56. The immunogenic composition according to claim 52, further comprising at least one additional antigen and/or at least one nucleic acid selected from nucleic acids capable of expressing at least one additional antigen.

57. The immunogenic composition according to claim 52, wherein the nucleic acid molecule is at least 90% identical to SEQ ID NO:

15 or SEQ ID NO;

19 or a fragment thereof.

58. The immunogenic composition according to claim 52, wherein the nucleic acid molecule is at least 90% identical to ORF 1 of PCV-B (SEQ ID NO:

23) or a fragment thereof.

59. The immunogenic composition according to claim 52, wherein the nucleic acid molecule is at least 90% identical to ORF 2 of PCV-B (SEQ ID NO:

25) or a fragment thereof.

60. The immunogenic composition according to claim 52, wherein the nucleic acid molecule is at least 90% identical to ORF 3 of PCV-B (SEQ ID NO:

27) or a fragment thereof.

61. The immunogenic composition according to claim 52, further comprising at least one antigen selected from viral antigens and bacterial antigens and/or at least one nucleic acid selected from nucleic acids capable of expressing an antigen of at least one member selected from viruses and bacteria.

62. The immunogenic composition according to claim 52, further comprising at least one member selected from viral toxins, bacterial toxins, viral antigens, and bacterial antigens and/or at least one nucleic acid selected from nucleic acids capable of expressing an antigen of at least one toxin selected from viral toxins, bacterial toxins, viruses, and bacteria.

63. The immunogenic composition according to claim 62, wherein the antigen of the at least one bacteria is E. coli.

64. The immunogenic composition according to claim 52, further comprising at least one vector comprising at least one nucleic acid selected from nucleic acids capable of expressing an antigen of at least one toxin selected from viral toxins, bacterial toxins, viruses, and bacteria.

65. The immunogenic composition according to claim 64, wherein at least one vector is contained by E. coli.

66. The immunogenic composition according to claim 52, further comprising:

a first member selected from polypeptides of a type B porcine circovirus and nucleic acids that express at least one polypeptide of a type B porcine circovirus, and

a second member selected from polypeptides of a type B porcine circovirus and nucleic acids that express at least one polypeptide of a type B porcine circovirus,

wherein at least one of the first and second members is the vector that contains and expresses in vivo in a porcine host cell, a nucleic acid molecule or fragments thereof encoding a type B circovirus antigen.

67. The immunogenic composition according to claim 66, wherein the polypeptides of a type B porcine circovirus contain at least one epitope of a type B porcine circovirus.

68. The immunogenic composition according to claim 66, wherein the polypeptides of a type B circovirus are epitopes of a type B porcine circovirus.

69. The immunogenic composition according to claim 66, wherein at least one member of the first and second members is selected from polypeptides of a type B porcine circovirus, and wherein the other member is the vector that contains and expresses in vivo in a porcine host cell, a nucleic acid molecule or fragments thereof encoding a type B porcine circovirus antigen.

70. The immunogenic composition according to claim 69, wherein the polypeptides of a type B porcine circovirus contain at least one epitope of a type B porcine circovirus.

71. The immunogenic composition according to claim 70, wherein the polypeptides of a type B porcine circovirus are epitopes of a type B porcine circovirus.

72. The immunogenic composition according to claim 52, further comprising an adjuvant.

73. A method for inducing an immunological response against a porcine circovirus comprising administering to a porcine the immunogenic composition of claim 52.

74. An immunogenic composition according to claim 52, wherein the vector is capable of inducing an immunogenic reaction in a porcine host against a type B porcine circovirus.

75. An isolated nucleotide sequence capable of hybridizing to a porcine circovirus type B nucleotide sequence, wherein the nucleotide sequence comprises at least 8 consecutive nucleotides derived from, or complementary to, a porcine circovirus type B sequence depicted in FIG. 8 (SEQ ID NO:

15 and SEQ ID NO;

19).

76. The nucleotide sequence of claim 75, wherein said nucleotide sequence is at least 12 consecutive nucleotides in length.

77. The nucleotide sequence of claim 75, wherein said nucleotide sequence is at least 20 consecutive nucleotides in length.

78. The nucleotide sequence of claim 75, wherein said nucleotide sequence has at least a 80% identity to a porcine circovirus type B depicted in FIG. 8 (SEQ ID NO:

15 and SEQ ID NO;

     19), or a fragment thereof.

79. The nucleotide sequence of claim 78, wherein said identity is at least 90%.

80. The nucleotide sequence of claim 78, wherein said identity is at least 95%.

88. A vector comprising:

(a) a nucleotide sequence according to claim 75;

and (b) elements allowing the expression and/or secretion of said nucleotide sequence in a host cell.

90. A host cell transformed with the vector of claim 88.

92. A method of producing a recombinant porcine circovirus type B polypeptide comprising:

(a) providing a population of host cells according to claim 90;

and (b) culturing said population of cells under conditions allowing the expression of a transfected porcine circovirus type B nucleotide sequence.

94. A polypeptide produced by the method of claim 92.

115. A nucleic acid hybridization assay for detecting porcine circovirus type B homologous sequences in a biological sample comprising:

(a) contacting a biological sample with a nucleotide sequence according to claim 78 under conditions which allow the hybridization of the nucleotide sequence of claim 78 and porcine circovirus type B nucleic acid present in the biological sample; and

(b) detecting the hybrid complex formed containing the nucleotide sequence of claim 78.

116. The assay of claim 115, wherein said nucleotide sequence is labeled, and the complexes are detected by detection of the presence of the label.

117. The assay of claim 115, wherein said detecting comprises using two porcine circovirus type B nucleic acid specific probes selected from nucleic acids of FIG. 8 (SEQ ID NO:

15 and SEQ ID NO;

19) and reagents for an amplification reaction of DNA.

121. The assay of claim 115, wherein said detecting comprises using two porcine circovirus type B nucleic acid specific probes, wherein the two probes define a sequence capable of hybridizing to a porcine circovirus type B nucleotide sequence;

hybridizing said probes to said nucleotide sequence to form hybridization complexes;

amplifying said porcine circovirus type B homologous sequences using said hybridization complexes until a desired degree of amplification has been achieved; and

identifying the amplification procedure.

122. A vector comprising:

(a) a nucleotide sequence according to claim 78;

and (b) elements operably linked to said nucleotide sequence in a host cell, whereby a coding sequence within said nucleotide sequence can be transcribed and translated in a host cell.

123. A probe or primer comprising all or part of a sequence of the nucleotide sequence of claim 75.

81. A nucleotide sequence encoding an immunogenic porcine circovirus type B polypeptide having at least 80% identity to a polypeptide selected from the group consisting of a polypeptide derived from (a) open reading frame (ORF)′

1 (SEQ ID NO;

23), (b) ORF′

2 (SEQ ID NO;

25), (c) ORF′

3 (SEQ ID NO;

27), and (d) immunogenic fragments of (a)-(c) comprising at least 5 consecutive amino acids.

82. The nucleotide sequence of claim 81 wherein (d) the immunogenic fragments of (a)-(c) comprise at least 10 consecutive amino acids.

83. The nucleotide sequence of claim 81, wherein (d) the immunogenic fragments of (a)-(c) comprise at least 15 consecutive amino acids.

84. The nucleotide sequence of claim 81, wherein said nucleotide sequence encodes an immunogenic porcine circovirus type B polypeptide having at least 90% identity to a polypeptide derived from ORF′

2 (SEQ ID NO;

25) and immunogenic fragments thereof comprising at least 5 consecutive amino acids.

85. The nucleotide sequence of claim 84, wherein the immunogenic fragments thereof comprise at least 10 consecutive amino acids.

86. The nucleotide sequence of claim 84, wherein the immunogenic fragments thereof comprise at least 15 consecutive amino acids.

87. The nucleotide sequence of claim 84, wherein the nucleotide sequence encodes the polypeptide of ORF′

2 (SEQ ID NO;

25).

89. A vector comprising:

(a) a nucleotide sequence according to claim 81;

and (b) elements allowing the expression and/or secretion of said nucleotide sequence in a host cell.

91. A host cell transformed with the vector of claim 89.

93. A method of producing a recombinant porcine circovirus type B polypeptide comprising:

(a) providing a population of host cells according to claim 91;

and (b) culturing said population of cells under conditions allowing the expression of a transfected porcine circovirus type B nucleotide sequence.

95. A polypeptide produced by the method of claim 93.

124. A probe or primer comprising all or part of a sequence of the nucleotide sequence of claim 81.

96. An immunogenic porcine circovirus type B polypeptide having at least 80% identity to a polypeptide selected from the group consisting of a polypeptide derived from (a) open reading frame (ORF)′

1 (SEQ ID NO;

23), (b) ORF′

2 (SEQ ID NO;

25), (c) ORF′

3 (SEQ ID NO;

27), and (d) immunogenic fragments of (a)-(c) comprising at least 5 consecutive amino acids.

97. The immunogenic porcine circovirus type B polypeptide of claim 96, wherein the identity is at least 90%.

98. The immunogenic porcine circovirus type B polypeptide of claim 96, wherein the polypeptide has at least 80% identity to a polypeptide derived from ORF′

2 (SEQ ID NO;

25), or immunogenic fragments thereof comprising at least 5 consecutive amino acids.

99. The immunogenic porcine circovirus type B polypeptide of claim 97, wherein the identity is at least 90%.

100. The immunogenic porcine circovirus type B polypeptide of claim 96, wherein the polypeptide has the sequence of the polypeptide encoded by ORF′

2 (SEQ ID NO;

25).

101. Antibodies that specifically recognize the polypeptide of claim 96.

102. Antibodies that specifically recognize the polypeptide of claim 97.

103. A composition comprising an immunogenic porcine circovirus type B polypeptide according to claim 96 and a pharmaceutically acceptable vehicle.

104. A composition comprising an immunogenic porcine circovirus type B polypeptide according to claim 97 and a pharmaceutically acceptable vehicle.

105. A method of producing a composition comprising providing an immunogenic porcine circovirus type B polypeptide according to claim 96 and combining said polypeptide with a pharmaceutically acceptable vehicle.

106. A method of producing a composition comprising providing an immunogenic porcine circovirus type B polypeptide according to claim 97 and combining said polypeptide with a pharmaceutically acceptable vehicle.

107. A method of treating or preventing porcine circovirus type B infection in a mammalian subject comprising administering to said subject a therapeutically effective amount of a composition according to claim 103.

108. A method of treating or preventing porcine circovirus type B infection in a mammalian subject comprising administering to said subject a therapeutically effective amount of a composition according to claim 104.

109. A method of detecting porcine circovirus type B antibodies in a biological sample comprising:

(a) providing a biological sample;

(b) contacting said biological sample with porcine circovirus type B polypeptide according to claim 96, under conditions allowing an immunological reaction between said polypeptide and the antibodies possibly present in said biological sample; and

(c) detecting the presence or absence of an antigen-antibody complex thereby detecting porcine circovirus type B antibodies in a biological sample.

110. A method of detecting porcine circovirus type B antibodies in a biological sample comprising:

(a) providing a biological sample;

(b) contacting said biological sample with porcine circovirus type B polypeptide according to claim 97, under conditions allowing an immunological reaction between said polypeptide and the antibodies possibly present in said biological sample; and

(c) detecting the presence or absence of an antigen-antibody complex thereby detecting porcine circovirus type B antibodies in a biological sample.

111. An immunodiagnostic test kit for detecting porcine circovirus type B infection in a animal subject, said test kit comprising an immunogenic porcine circovirus type B polypeptide according to claim 96.

112. An immunodiagnostic test kit for detecting porcine circovirus type B infection in a animal subject, said test kit comprising an immunogenic porcine circovirus type B polypeptide according to claim 97.

113. An immunodiagnostic test kit of claim 111, wherein the animal subject is a mammalian subject.

114. An immunodiagnostic test kit of claim 112, wherein the animal subject is a mammalian subject.

118. An immunodiagnostic test kit for detecting porcine circovirus type B infection in an animal, said test kit comprising the antibody according to claim 101.

119. An immunodiagnostic test kit for detecting porcine circovirus type B infection in an animal, said test kit comprising the antibody according to claim 102.

120. The kit of claim 118, wherein said animal is a mammal.

125. A composition for modulating cellular replication of porcine circovirus type B (PCV-B) comprising a pharmaceutically or veterinarily acceptable vehicle and an active agent comprising a PCV-B immunogen present in an amount effective to modulate cellular replication, wherein said immunogen is an inactivated PCV-B.

126. The composition of claim 125, wherein the immunogen is recombinantly produced.

127. The composition of claim 125, wherein the immunogen comprises a polypeptide encoded by a sequence having at least 80% identity to SEQ ID NO:

15 or SEQ ID NO;

19, or fragments thereof.

128. The composition of claim 127, wherein the immunogen comprises a polypeptide encoded by a sequence having at least 90% identity to SEQ ID NO:

15 or SEQ ID NO;

19, or fragments thereof.

129. The composition of claim 128, wherein the immunogen comprises a polypeptide encoded by a sequence having at least 95% identity to SEQ ID NO:

15 or SEQ ID NO;

19, or fragments thereof.

130. The composition of claim 125, wherein the immunogen comprises a polypeptide encoded by a sequence having at least 80% identity to SEQ ID NO:

25, or a fragment thereof.

131. The composition of claim 130, wherein the immunogen comprises a polypeptide encoded by a sequence having at least 90% identity to SEQ ID NO:

25, or a fragment thereof.

132. The composition of claim 131, wherein the immunogen comprises a polypeptide encoded by a sequence having at least 95% identity to SEQ ID NO:

25, or a fragment thereof.

133. A method for modulating cellular replication of porcine circovirus type B (PCV-B) in a pig comprising administering to the pig the composition of claim 125.

134. A method for modulating cellular replication of porcine circovirus type B (PCV-B) in a pig comprising administering to the pig the composition of claim 126.

135. A method for modulating cellular replication of porcine circovirus type B (PCV-B) in a pig comprising administering to the pig the composition of claim 127.

136. A method for modulating cellular replication of porcine circovirus type B (PCV-B) in a pig comprising administering to the pig the composition of claim 128.

137. A method for modulating cellular replication of porcine circovirus type B (PCV-B) in a pig comprising administering to the pig the composition of claim 129.

138. A method for modulating cellular replication of porcine circovirus type B (PCV-B) in a pig comprising administering to the pig the composition of claim 130.

139. A method for modulating cellular replication of porcine circovirus type B (PCV-B) in a pig comprising administering to the pig the composition of claim 131.

140. A method for modulating cellular replication of porcine circovirus type B (PCV-B) in a pig comprising administering to the pig the composition of claim 132.

141. The method of claim 133, wherein the immunogen is recombinantly produced.

142. The method of claim 133, wherein the administering is prior to breeding.

143. The method of claim 133, wherein the administering is to a pregnant female pig.

144. A composition for modulating viral replication of porcine circovirus type B (PCV-B) comprising a pharmaceutically or veterinarily acceptable vehicle and an active agent comprising a PCV-B immunogen present in an amount effective to modulate cellular replication, wherein said immunogen is an inactivated PCV-B.

145. The composition of claim 144, wherein the immunogen is recombinantly produced.

146. The composition of claim 144, wherein the immunogen comprises a polypeptide encoded by a sequence having at least 80% identity to SEQ ID NO:

15 or SEQ ID NO;

19, or fragments thereof.

147. The composition of claim 146, wherein the immunogen comprises a polypeptide encoded by a sequence having at least 90% identity to SEQ ID NO:

15 or SEQ ID NO;

19, or fragments thereof.

148. The composition of claim 147, wherein the immunogen comprises a polypeptide encoded by a sequence having at least 95% identity to SEQ ID NO:

15 or SEQ ID NO;

19, or fragments thereof.

149. The composition of claim 144, wherein the immunogen comprises a polypeptide encoded by a sequence having at least 80% identity to SEQ ID NO:

25, or a fragment thereof.

150. The composition of claim 149, wherein the immunogen comprises a polypeptide encoded by a sequence having at least 90% identity to SEQ ID NO:

25, or a fragment thereof.

151. The composition of claim 150, wherein the immunogen comprises a polypeptide encoded by a sequence having at least 95% identity to SEQ ID NO:

25, or a fragment thereof.

152. A method for modulating cellular replication of porcine circovirus type B (PCV-B) in a pig comprising administering to the pig the composition of claim 144.

153. A method for modulating cellular replication of porcine circovirus type B (PCV-B) in a pig comprising administering to the pig the composition of claim 145.

154. A method for modulating cellular replication of porcine circovirus type B (PCV-B) in a pig comprising administering to the pig the composition of claim 146.

155. A method for modulating cellular replication of porcine circovirus type B (PCV-B) in a pig comprising administering to the pig the composition of claim 147.

156. A method for modulating cellular replication of porcine circovirus type B (PCV-B) in a pig comprising administering to the pig the composition of claim 148.

157. A method for modulating cellular replication of porcine circovirus type B (PCV-B) in a pig comprising administering to the pig the composition of claim 149.

158. A method for modulating cellular replication of porcine circovirus type B (PCV-B) in a pig comprising administering to the pig the composition of claim 150.

159. A method for modulating cellular replication of porcine circovirus type B (PCV-B) in a pig comprising administering to the pig the composition of claim 151.

160. The method of claim 144, wherein the immunogen is recombinantly produced.

161. The method of claim 144, wherein the administering is prior to breeding.

162. The method of claim 144, wherein the administering is to a pregnant female pig.