Human interleukin-4 antagonist/agonist screens
AssigneeSchering Corporation
InventorsJan E. de Vries, Chung-Her Jenh, Satwant K. Narula, Paul J. Zavodny
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Abstract
Stably-transformed human cell lines containing a reporter gene operatively linked to an IL-4-responsive element are provided. In some embodiments a human Fc.sub..epsilon. RII IL-4-responsive element is used. In other embodiments a human germline .epsilon. transcript promoter is used as the responsive element. Also provided are methods for using such transformed cell lines to screen for agonists and/or antagonists of human interleukin-4.
Claims18
A method for identifying a compound in a sample as a human IL-4 agonist comprising:
- (a) providing a cell line expressing receptors for human IL-4, which cell line has been stably transfected by a recombinant vector comprising a reporter gene operatively linked to a human Fc.sub..epsilon. RII IL-4 responsive element comprising nucleotides 298-507 of SEQ ID NO:
- 1, or to a germline .epsilon. transcript promoter comprising nucleotides 7-587 of SEQ ID NO:
- 2, which responsive element or promoter is capable of controlling expression of the reporter gene in response to IL-4;
- (b) contacting the transfected cell line with a sample suspected to contain a human IL-4 agonist, under conditions in which human IL-4 would cause increased expression of the reporter gene;
- and
- (c) measuring the level of expression of the reporter gene,
- whereby a human IL-4 agonist in the sample is identified by measurement of an increased level of expression of the reporter gene, compared to the level produced by a buffer control.
A method for identifying a compound in a sample as a human IL-4 antagonist comprising:
- (a) providing a cell line expressing receptors for human IL-4, which cell line has been stably transfected by a recombinant vector comprising a reporter gene operatively linked to a human Fc.sub..epsilon. RII IL-4 responsive element comprising nucleotides 298-507 of SEQ ID NO:
- 1, or to a germline .epsilon. transcript promoter comprising nucleotides 7-587 of SEQ ID NO:
- 2, which responsive element or promoter is capable of controlling expression of the reporter gene in response to IL-4;
- (b) contacting the transfected cell line with a sample suspected to contain a human IL-4 antagonist, to which has been added an amount of human IL-4 that, absent such antagonist, would produce a measurable increase in expression of the reporter gene;
- and
- (c) measuring the level of expression of the reporter gene,
- whereby a human IL-4 antagonist in the sample is identified by measurement of a decreased level of expression of the reporter gene, compared to the level produced by the human IL-4 in the absence of such antagonist.
Dependencies are inferred from the claim text.