Identification of agents that protect against inflammatory injury to neurons
AssigneeBaylor College of Medicine
InventorDana Giulian
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Abstract
The present invention is directed to screening for an agent that inhibits the effect of a neurotoxin from a plaque component activated mononuclear phagocyte on a neuron. In addition, the present invention is directed to methods of screening for agents that inhibit mononuclear phagocyte-plaque component complex formation, plaque component activation of a mononuclear phagocyte, plaque component induced neurotoxicity of a mononuclear phagocyte. An agent obtained by the method of screening for an agent that inhibits mononuclear phagocyte-plaque component complex formation and a pharmaceutical composition comprising the agent are embodied by the present invention.
Claims28
A method of screening for an agent that inhibits the activation of a mononuclear phagocyte by complex formation of β-amyloid or active fragments of β-amyloid and the mononuclear phagocyte, wherein said active fragments at a concentration of from about 0.01 to about 10 μmole/liter induce significant reduction in neuron survival as defined by loss of neuron specific markers in neuron cultures containing microglia compared to identical cultures without the presence of β-amyloid or active fragments thereof said method comprising:
- contacting one or more mononuclear phagocytes with said β-amyloid or said active Moments of β-amyloid effective to form a complex and to activate said mononuclear phagocyte, and an agent suspected of inhibiting said activation,
- measuring ex vivo said activation of said mononuclear phagocyte in the presence of the agent, and
- comparing said measured activation of said mononuclear phagocyte to activation of the mononuclear phagocyte in the absence of the agent,
- wherein a reduction in activation of said mononuclear phagocyte in the presence of the agent compared to activation of the mononuclear phagocyte in the absence of the agent is indicative of an agent that inhibits activation of said mononuclear phagocyte by β-amyloid or active fragments thereof and further wherein said activation is measured by imaging the mononuclear phagocyte.
A method of screening for an agent that inhibits the activation of a mononuclear phagocyte by complex formation of β-amyloid or active fragments of β-amyloid and the mononuclear phagocyte, wherein said active fragments at a concentration of from about 0.01 to about 10 μmole/liter induce significant reduction in neuron survival as defined by loss of neuron specific markers in neuron cultures containing microglia compared to identical cultures without the presence of β-amyloid or active fragments thereof, said method comprising:
- contacting one or more mononuclear phagocytes with β-amyloid or active fragments of β-amyloid effective to form a complex and to activate said mononuclear phagocyte, wherein said β-amyloid or active fragments are coupled to a solid support, and further contacting said mononuclear phagocytes with an effective amount of an agent suspected of inhibiting said activation,
- measure ex vivo said activation of said mononuclear phagocyte in the presence of the agent, and
- comparing said measured activation of said mononuclear phagocyte to activation of the mononuclear phagocyte in the absence of the agent,
- wherein a reduction in activation of said mononuclear phagocyte in the presence of the agent compared to activation of the mononuclear phagocyte in the absence of the agent is indicative of an agent that inhibits activation of said mononuclear phagocyte by β-amyloid or active fragments thereof, and further wherein said activation is measured by imaging the mononuclear phagocyte.
A method of screening for an agent that inhibits the activation of a mononuclear phagocyte by complex formation of β-amyloid or active fragments of β-amyloid and the mononuclear phagocyte, wherein said active fragments at a concentration of from about 0.01 to about 10 μmole/liter induce significant reduction in neuron survival as defined by loss of neuron specific markers in neuron cultures containing microglia compared to identical cultures without the presence of β-amyloid or active fragments thereof, said method comprising:
- contacting one or more mononuclear phagocytes in a mammalian brain with β-amyloid or active fragments of β-amyloid effective to form a complex and to activate said mononuclear phagocyte, wherein said β-amyloid or active fragments are coupled to a solid support, and further contacting the mononuclear phagocyte with an agent suspected of inhibiting said activation,
- measuring ex vivo said activation of said mononuclear phagocyte in the presence of the agent, and
- comparing said measured activation of said mononuclear phagocyte to activation of the mononuclear phagocyte in the absence of the agent,
- wherein a reduction in activation of said mononuclear phagocyte in the presence of the agent compared to activation of the mononuclear phagocyte in the absence of the agent is indicative of an agent that inhibits activation of said mononuclear phagocyte by β-amyloid or active fragments thereof and wherein said activation is measured by imaging the mononuclear phagocyte.
Dependencies are inferred from the claim text.