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US 2004/0106104 A1Published application
Viral envelope mediated fusion assay
OwnerPin-Fang Lin, Carol Deminie +3inventors
InventorsPin-Fang Lin, Carol Deminie, Robert A. Fridell, Brett Robinson, Ronald E. Rose
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Abstract
This invention provides a method of screening a sample for susceptibility to a viral entry inhibitor. The invention further provides a method for monitoring a patient for sensitivity to a viral entry inhibitor and a method of determining an inhibitory concentration of a viral entry inhibitor to inhibit virus infection in a patient. A method of screening for viral entry inhibitors is also provided.
Claims45
A method of screening a sample for sensitivity to a viral entry inhibitor comprising the steps of:
- (a) culturing in the presence of a viral entry inhibitor (i) an effector expressing a linear viral envelope expression element and a transcription factor with (ii) a target cell capable of fusing with the effector in the absence of the viral entry inhibitor, said target cell containing a nucleic acid encoding a reporter wherein said nucleic acid is capable of expressing the reporter in response to the transcription factor of the effector;
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- (b) detecting expression of the reporter wherein expression of the reporter is indicative of sensitivity of the sample to the viral entry inhibitor.
A method for monitoring a patient for sensitivity to a viral entry inhibitor comprising the steps of:
- (a) culturing in the presence of a viral entry inhibitor (i) a first effector expressing a linear viral envelope expression element prepared from a sample obtained from the patient at a first time and a transcription factor with (ii) a first target cell capable of fusing with the first effector in the absence of the viral entry inhibitor, said first target cell containing a nucleic acid encoding a reporter wherein said nucleic acid is capable of expressing the reporter in response to the transcription factor of the first effector;
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- (b) culturing in the presence of the viral entry inhibitor (i) a second effector expressing a linear viral envelope expression element prepared from a sample obtained from the patient at a second time and a transcription factor with (ii) a second target cell capable of fusing with the second effector in the absence of the viral entry inhibitor, said second target cell containing a nucleic acid encoding a reporter wherein said nucleic acid is capable of expressing the reporter in response to the transcription factor of the second effector;
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- (c) determining whether there is a change in expression of the reporter in the second target cell when compared to the expression of the reporter in the first target cell wherein an increase in expression of the reporter in the second target cell compared to the expression of the reporter in the first target cell indicates decreased sensitivity of the patient to the viral entry inhibitor.
2 more elements
A method for identifying viral entry inhibitors comprising the steps of:
- (a) culturing in the presence of a candidate viral entry inhibitor (i) an effector cell expressing a linear viral envelope expression element and a transcription factor with (ii) a target cell capable of fusing with the effector cell in the absence of the viral entry inhibitor, said target cell containing a nucleic acid encoding a reporter wherein said nucleic acid is capable of expressing the reporter in response to the transcription factor of the effector cell;
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- (b) determining whether there is a reduction in expression of the reporter in the presence of the candidate viral entry inhibitor when compared to the expression of the reporter in the absence of the candidate viral entry inhibitor wherein a reduction in expression of the reporter in the presence of the candidate viral entry inhibitor when compared to the expression of the reporter in the absence of the candidate viral entry inhibitor indicates that the candidate viral entry inhibitor is a viral entry inhibitor.
A method of determining an inhibitory concentration of a viral entry inhibitor to inhibit virus infection in a patient comprising the steps of:
- (a) culturing in the presence of a concentration of viral entry inhibitor (i) an effector expressing a linear viral envelope expression element and a transcription factor with (ii) a target cell capable of fusing with the effector in the absence of the viral entry inhibitor, the target cell containing a nucleic acid encoding a reporter wherein the nucleic acid is capable of expressing the reporter in response to the transcription factor of the effector;
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- (b) determining the concentration of viral entry inhibitor that inhibits a desired percentage of viral replication of the virus.
A method of screening a sample for sensitivity to a viral entry inhibitor comprising the steps of:
- (a) culturing in the presence of a viral entry inhibitor (i) a pseudovirus produced from the expression of a linear viral envelope expression element, wherein the pseudovirus includes a reporter, and (ii) a target cell capable of fusing with the pseudovirus in the absence of the viral entry inhibitor, wherein the target cell is capable of expressing the reporter of the pseudovirus;
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- (b) detecting expression of the reporter wherein expression of the reporter is indicative of sensitivity of the sample to the viral entry inhibitor.
A method for identifying viral entry inhibitors comprising the steps of:
- (a) culturing in the presence of a candidate viral entry inhibitor (i) a pseudovirus produced from the expression of a linear viral envelope expression element, wherein the pseudovirus includes a reporter, and (ii) a target cell capable of fusing with the pseudovirus in the absence of the viral entry inhibitor, wherein the target cell is capable of expressing the reporter of the pseudovirus;
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- (b) determining whether there is a reduction in expression of the reporter in the presence of the candidate viral entry inhibitor when compared to the expression of the reporter in the absence of the candidate viral entry inhibitor wherein a reduction in expression of the reporter in the presence of the candidate viral entry inhibitor when compared to the expression of the reporter in the absence of the candidate viral entry inhibitor indicates that the candidate viral entry inhibitor is a viral entry inhibitor.
Dependencies are inferred from the claim text.