Methods and cell lines for screening compositions and genes for ability to interact with IL-1β and ice processing

A method for identifying a composition with activity in Interleukin-1β (IL-1β) processing or secretory pathways comprising:

• providing a non-naturally occurring cell that has been modified to comprise a first gene for precursor IL-1β (preIL-β) operably linked to a promoter, and a second gene selected from the group consisting of:
• (a) a gene for Interleukin-1β Converting Enzyme (ICE), (b) a gene for an ICE-like molecule capable of processing preIL-β to mature IL-β (matIL-β) and (c) a gene for a test composition to be evaluated, the second gene being operably linked to a promoter;
• The first and second genes being stably incorporated into the cell;
• culturing the cell at a temperature of 29° C. to 33° C., under conditions and for a period of time sufficient for expression of the first and second genes by the cell;
• applying to the cell a test composition to be evaluated when the second gene is a gene for ICE or an ICE-like molecule capable of processing preIL-β to matIL-β;
• and
• monitoring the cell or the cell environment for a change in the concentration of preIL-1β or matIL-1β, as compared to the concentration of preILβ or matIL-1β in the absence of the test composition, which change identifies the test composition as a composition having activity in IL-1β processing and secretory pathways.

A method for identifying a composition with activity in Interleukin-1β (IL-1β) processing or secretory pathways comprising:

• providing a non-naturally occurring cell that has been modified to comprise a first gene for precursor IL-1β (preIL-β) operably linked to a promoter, and a second gene selected from the group consisting of:
• (a) a gene for Interleukin-1β Converting Enzyme (ICE), (b) a gene for an ICE-like molecule capable of processing preIL-β to mature IL-β (matIL-β) and (c) a gene for a test composition to be evaluated, the second gene being operably linked to a promoter;
• the first and second genes being stably incorporated into the cell;
• culturing the cell a a temperature of 31° C. to 33° C., under conditions and for a period of time sufficient for expression of the first and second genes by the cell;
• applying to the cell a test composition to be evaluated when the second gene is a gene for ICE or an ICE-like molecule capable of processing preIL-β to matIL-β;
• and
• monitoring the cell or the cell environment for a change in the concentration of preIL-1β or matlL-1β, as compared To the concentration of preILβ or matIL-1β in the absence of the test composition, which change identifies the test composition as a composition having activity in IL-1β processing and secretory pathways.

A method for identifying a modulator of Interleukin-1 β Converting Enzyme (ICE) or an ICE-like molecule, comprising:

• providing a non-naturally occurring cell that has been modified to comprise a first gene for ICE or an ICE-like molecule and a second gene for a substrate of ICE or the ICE-like molecule, the substrate being processed from a precursor form to a mature form by ICE or the ICE-like molecule;
• the first and second genes being stably incorporated into the cell;
• culturing the cell at a temperature of 31° C. to the 33° C., under conditions and for a period of time sufficient for expression of the first and second genes by the cell;
• applying to the cell a test composition to be evaluated;
• and
• monitoring the cell or the cell environment for a change in the concentration of the precursor form or mature form of the substrate, as compared to the concentration of the precursor form or mature form of the substrate in the absence of the test composition, which change identifies the test composition as a modulator of ICE or the ICE-like molecule.

A method for identifying a modulator of Interleukin-1β Converting Enzyme (ICE) or an ICE-like molecule, comprising:

• providing a non-naturally occurring cell that has been modified to comprise a first gene for ICE or an ICE-like molecule, wherein the cell naturally expresses a second gene for a substrate of ICE or the ICE-like molecule, the substrate being processed from a precursor form to a mature form by ICE or the ICE-like molecule;
• the first and second genes being stably incorporated into the cell;
• culturing the cell at a temperature of 31° C. to 33° C., under conditions and for a period of time sufficient for expression of the first and second genes by the cell;
• applying to the cell a test composition to be evaluated;
• and
• monitoring the cell or the cell environment for a change in the concentration of the precursor form or mature form of the substrate, as compared to the concentration of the precursor form or mature form of the substrate in the absence of to test composition, which change identifies the test composition as a modulator of ICE or the ICE-like molecule.

A method for identifying a modulator of Interleukin-1β Converting Enzyme (ICE) or an ICE-like molecule, comprising:

• providing a non-naturally occurring cell that has been modified to comprise a first gene for ICE or an ICE-like molecule, wherein the cell naturally expresses a second gene for precursor IL-1β (preIL-1β);
• the first and second genes being stably incorporated into the cell;
• culturing the cell at a temperature of 31° C. to 33° C., under conditions and for a period of time sufficient for expression of the first and second genes by the cell;
• applying to the cell a test composition to be evaluated;
• and
• monitoring the cell or the cell environment for a change in the concentration of preIL-1β or matIL-1β, as compared to be concentration of preIL-1 β or matIL-1β in the absence of the test composition, which change identifies the test composition as a modulator of ICE or the ICE-like molecule.