Cellular virus receptors and methods of use

• US 20050101530A1
• Filed: 08/12/2002
• Published: 05/12/2005
• Est. Priority Date: 08/10/2001
• Status: Abandoned Application

• Current Assignee: Topigen Pharmaceuticals, Inc. (Syntara Ltd.)
• Original Assignee: Topigen Pharmaceuticals, Inc. (Syntara Ltd.)
• Inventors: Renzi, Paolo, Zemzoumi, Khalid
• Application Number:
  US10/486,471
• Publication Number:
  US 20050101530A1
• Time in Patent Office: Days
• US Class Current: 435/5
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 38/1793 for cytokines; for lymphokines; for interferons
• CPC Class Codes: A61K 38/195 Chemokines, e.g. RANTES
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: C07K 14/7155 for interleukins [IL]
• CPC Class Codes: C07K 16/1027 Paramyxoviridae, e.g. respiratory syncytial virus
• CPC Class Codes: C07K 16/2866 against receptors for cytokines, lymphokines, interferons
• CPC Class Codes: C07K 2317/32 specific for a neo-epitope on a complex, e.g. antibody-antigen or ligand-receptor
• CPC Class Codes: C07K 2317/76 Antagonist effect on antigen, e.g. neutralization or inhibition of binding
• CPC Class Codes: G01N 2333/715 for cytokines; for lymphokines; for interferons
• CPC Class Codes: G01N 2500/00 Screening for compounds of potential therapeutic value
• CPC Class Codes: G01N 33/56983 Viruses

Abstract

Methods, reagents and compositions for the treatment, prevention and diagnotic of virus infections in vertebrates and more paticularly in human and animals are described. The invention provides evidence that the CCR1, CCR2, CCR3, CCR4, CCR5 and CCR8 receptors are involved in human respiratory syncytial virus (RSV) infections. Therefore, the present invention describes methods for modulation of cellular viral infection by modulating a binding interaction between a CCR1, CCR2, CCR3, CCR4, CCR5 and/or CCR8 receptor and a surface protein of the virus. The invention also profits of such a binding interaction for 10 providing methods for reducing viral infection of a cell; methods of attenuating the ability of a pneumovirus to bind a mammalian cell; methods for reducing the initiation or spread of a respiratory tract disease due to human RSV; methods for detecting the presence of a pneumovirus in a biological sample; gene therapy methods, and methods for identifying novel antiviral and anti-inflammatory 15 compounds.

Claims

1-85. -85. (canceled)

86. A method for modulating viral infection of a cell, comprising modulating a binding interaction between a cell chemokine-receptor and a surface protein of said virus, said cell chemokines-receptor comprising an amino acid sequence having at least 38% identity with SEQ ID NO:

6, with the proviso that said virus is not HIV.

87. The method of claim 86, wherein said cell chemokine-receptor is selected from the group consisting of CCR1, CCR2, CCR3, CCR4, CCR5, and CCR8.

88. The method of claim 87, wherein said cell chemokine-receptor consists of CCR3.

89. A method of modulating a pneumovirus infection of a cell, comprising modulating a binding interaction between at least one chemokine-receptor of said cell and a surface protein of said pneumovirus, wherein said cell chemokine-receptor is selected from the group consisting of CCR1, CCR2, CCR3, CCR4, CCR5, and CCR8.

90. The method of claim 89, comprising the step of contacting said cell with a CCR-ligand that recognizes at least one of said cell chemokine-receptor, said contacting step being carried out under conditions sufficient for said CCR-ligand to bind at least one of said cell chemokine-receptor, whereby the ability of said pneumovirus to subsequently bind said cell is attenuated.

91. The method of claim 90, wherein said CCR-ligand is selected form the group consisting of:

eotaxin I, eotaxin 2, eotaxin 3, monocyte chemoattractant protein-1 (MCP-1), MCP-2, MCP-3, MCP-4, MCP-5, MIP-1α

, MIP-1β

, I-309, regulated on activation normal T-cell expressed and secreted (RANTES), TARC, antibodies and fragments thereof which specifically recognize said cell receptor(s), and polypeptides having a binding activity to an extracellular domain of said receptor(s).

92. The method of claim 91, comprising the step of reducing intracellular levels of at least one of said cell chemokine-receptor with antisense molecules introduced or expressed into said cell, thereby limiting pneumovirus ability to infect said cell.

93. The method of claim 89, comprising the step of contacting said pneumovirus with a virus-ligand that recognizes a surface protein of said pneumovirus under conditions sufficient for said virus-ligand to bind said pneumovirus surface protein and subsequently attenuates binding of said pneumovirus to said cell chemokine-receptor(s).

94. The method of claim 93, wherein said virus-ligand is selected from the group consisting of:

a. an antibody or fragment thereof which specifically recognizes said virus surface protein; and

b. a CCR1, CCR2, CCR3, CCR4, CCR5 and/or CCR8 receptor or a functional fragment thereof.

95. The method of claim 94, wherein said antibody or fragment thereof specifically binds to a RSV-G or RSV-F protein.

96. The method of claim 95, wherein said virus consists of the Human Respiratory Syncytial Virus (HRSV).

97. A method of reducing the initiation or spread of a respiratory tract disease due to human RSV, comprising administering to a human an antiviral agent comprising a virus-ligand which exhibits the ability to bind to HRSV and reduce infectivity thereof, said virus-ligand exhibiting the ability to bind to a HRSV surface protein involved in binding of the HRSV to a cell chemokine-receptor selected from the group consisting of CCR1, CCR2, CCR3, CCR4, CCR5, and CCR8.

98. A method for detecting the presence of a pneumovirus in a biological sample, comprising the steps of:

a. contacting the biological sample with a cell expressing at least one cell chemokine-receptor selected from the group consisting of CCR1, CCR2, CCR3, CCR4, CCR5, and CCR8, said contacting step being carried out under conditions sufficient for pneumovirus(es) contained in said sample to infect said cell; and

b. detecting the presence of pneumovirus(es) in said cell.

99. The method of claim 98, wherein said cell is selected from the group consisting of:

Hep-2 cells, A549 cells, and GHOST cells transfected with the CCR1, CCR2, CCR3, CCR4, CCR5 and/or CCR8 receptor(s).

100. The method of claim 99, wherein said cell has been pretreated with agent(s) capable of increasing or permitting the expression of said chemokine-receptor(s) on the cell surface.

101. A method for selecting an antiviral compound that is capable of reducing viral infection of a cell by a pneumovirus, the cell expressing at least one cell chemokine-receptor selected from the group consisting of CCR1, CCR2, CCR3, CCR4, CCR5 and CCR8, the method comprising:

a. contacting a functional virus-ligand and a surface protein of a virus in the presence of a potential antiviral compound, said surface protein being involved in binding of the virus to at least one of the chemokine-receptor expressed by the cell;

b. measuring a binding interaction between said virus-ligand and said viral surface protein;

whereby an antiviral compound is selected when said binding interaction is measurably reduced in presence of said potential antiviral compound.

102. The method of claim 101, wherein said virus-ligand is selected from the group consisting of:

a. an antibody or a fragment thereof which specifically recognizes said pneumovirus surface protein; and

b. a CCR1, CCR2, CCR3, CCR4, CCR5 and/or CCR8 receptor or a functional fragment thereof.

103. The method of claim 102, wherein said pneumovirus consists of the Human Respiratory Syncytial Virus (HRSV).

104. A composition for treating and/or preventing infection by a pneumovirus, said composition comprising a CCR-ligand and a pharmaceutically acceptable carrier, said CCR-ligand exhibiting the ability to bind to at least one chemokine-receptor selected from the group consisting of CCR1, CCR2, CCR, CCR4, CCR5, and CCR8, and thereby reduce infectivity of the pneumovirus.

105. The composition of claim 104, wherein said composition consists of a spray for application to airway tissues susceptible to infection by HRSV.

106. A method of attenuating the ability of a pneumovirus to infect a mammalian cell expressing at least one chemokine-receptor selected from the group consisting of CCR1, CCR2, CCR3, CCR4, CCR5, and CCR8, the method comprising exposing said cell to a CCR-ligand that recognizes at least one of said cell chemokine-receptor, said exposition being carried out under conditions sufficient for said CCR-ligand to bind said at least one of said cell chemokine-receptor, whereby the ability of said pneumovirus to subsequently infect said cell is attenuated.

107. The method of claim 106, wherein said CCR-ligand is selected from the group consisting of:

eotaxin 1, eotaxin 2, eotaxin 3, monocyte chemoattractant protein-1 (MCP-1), MCP-2, MCP-3, MCP-4, MCP-5, MIP-1α

, MIP-1β

, I-309, regulated on activation normal T-cell expressed and secreted (RANTES), TARC, antibodies and fragments thereof which specifically recognize said cell receptor(s), polypeptides having a binding activity to an extracellular domain of said receptor(s).

108. A method of attenuating the ability of a pneumovirus to bind a mammalian cell, comprising a step selected from the group consisting of:

a. exposing said cell to a CCR-ligand that recognizes at least one cell chemokine-receptor selected from the group consisting of CCR1, CCR2, CCR3, CCR4, CCR5, and CCR8, said exposition being carried out under conditions sufficient for said CCR-ligand to bind said cell receptor;

b. exposing said virus to a virus-ligand that recognizes a surface protein of the pneumovirus, said exposition being carried out under conditions sufficient for said virus-ligand to bind said pneumovirus surface protein and subsequently attenuates binding of pneumovirus to at least one cell chemokine-receptor selected from the group consisting of CCR1, CCR2, CCR3, CCR4, CCR5, and CCR8; and

c. reducing intracellular levels of at least one cell chemokine-receptor selected from the group consisting of CCR1, CCR2, CCR3, CCR4, CCR5, and CCR8;

thereby limiting the pneumovirus ability to bind said mammalian cell.