Differential regulation of T cell survival and proliferation
AssigneeWashington University
InventorsJonathan M. Green, Andrey S. Shaw
Life of this patent
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Abstract
The present invention provides methods for selectively regulating survival and proliferation in a T cell by modulating the activity of certain amino acid motifs in the CD28 protein.
Claims59
A method for selectively modulating T cell survival relative to T cell proliferation comprising contacting a T cell expressing a CD28 protein with an agent that selectively modulates the activity of a CD28 survival motif relative to the activity of a CD28 proliferation motif, to thereby selectively modulate survival of the T cell.
The method of claiml, wherein said modulation of T cell survival is reduction of T cell survival.
A method for treating a subject having a condition that would benefit from selective enhancement of T cell survival relative to T cell proliferation comprising,
- a) providing an agent that selectively increases the activity of a CD28 survival motif relative to the activity of a CD28 proliferation motif;
- and
- b) administering the agent to the subject, to thereby selectively increase survival of the T cells of the subject relative to proliferation of the T cells of the subject.
A method for treating a subject having a condition that would benefit from selective reduction of T cell survival relative to T cell proliferation, comprising:
- a) providing an agent that selectively decreases the activity of a CD28 survival motif relative to the activity of a CD28 proliferation motif;
- and
- b) administering the agent to the subject to deliver the agent intracellularly to T cells of the subject, to thereby selectively decrease survival of the T cells of the subject relative to proliferation of the T cells of the subject.
A cell-based method for identifying a compound which selectively modulates T cell survival relative to T cell proliferation, comprising:
- a) providing a cell which expresses a CD28 protein that comprises a CD28 survival motif, in the context of a cell-based assay;
- b) contacting the cell of step a) with a test compound;
- and
- c) assaying for the ability of the test compound to modulate the phosphorylation of CD28 at a tyrosine residue which corresponds to the tyrosine at position 170 of the CD28 protein set forth in SEQ ID NO:2;
- wherein a determination that the test compound modulates the phosphorylation of the CD28 protein, indicates that the test compound is a compound which selectively modulates T cell survival.
A cell-based method for identifying a compound which selectively modulates T cell survival relative to T cell proliferation comprising:
- a) providing a cell which expresses a CD28 protein that comprises a CD28 survival motif, in the context of a cell based assay;
- b) contacting the cell of step a) with a test compound;
- and
- c) assaying for the ability of the test compound to modulate the activity or expression of Bcl-XL in the cell;
- wherein a determination that the test compound modulates the activity or expression of Bcl-XL in the cell indicates that the test compound is a compound which selectively modulates T cell survival.
A cell-free method for identifying a compound which selectively modulates T cell survival relative to T cell proliferation, comprising;
- a) providing a polypeptide comprising a CD28 survival motif in the context of a cell-free assay system;
- b) contacting the polypeptide with a test compound;
- and
- c) assaying for the ability of the test compound to modulate phosphorylation of the CD28 survival motif at a tyrosine that corresponds to the tyrosine at position 170 of CD28 set forth in SEQ ID NO:2, by measuring a readout selected from the group consisting of:
- (i) test compound binding to the region of the polypeptide comprising the tyrosine residue;
- and
- (ii) test compound modulation of phosphorylation of CD28 survival motif at the tyrosine residue;
- wherein detection of either (i) or (ii) indicates that the test compound is a compound that selectively modulates T cell survival.
A method for selectively modulating T cell proliferation relative to T cell survival comprising contacting a T cell expressing a CD28 protein with an agent that selectively modulates the activity of a CD28 proliferation motif relative to the activity of a CD28 survival motif, to thereby selectively modulate proliferation of the T cell relative to survival of the T cell.
A method for treating a subject having a condition that would benefit from selective increase of T cell proliferation relative to T cell survival comprising,
- a) providing an agent that acts intracellularly to induce or enhance interaction of the prolines of CD28 protein which correspond to the proline at position 187 and the proline at position 190 of the CD28 protein set forth in SEQ ID NO:2 with a molecule with which the CD28 proliferation motif interacts;
- and
- b) administering the agent to the subject to deliver the agent intracellularly to T cells of the subject, to thereby selectively increase proliferation of the T cells.
A method for treating a subject having a condition that would benefit from selective reduction of T cell proliferation relative to T cell survival comprising,
- a) providing an agent that acts intracellularly-to diminish or interfere with interaction of the prolines of the CD28 protein which correspond to the proline at position 187 and the proline at position 190 of the CD28 protein set forth in SEQ ID NO:2, with a molecule which interacts with a CD28 proliferation motif;
- and
- b) administering the agent of step a) to the subject to deliver the agent intracellularly to T cells of the subject, to thereby selectively reduce proliferation of the T cells.
A cell-based method for identifying a compound which selectively modulates T cell proliferation relative to T cell survival, comprising:
- a) providing a cell, which expresses a CD28 protein that comprises a CD28 proliferation motif, in the context of a cell-based assay;
- b) contacting the cell of step a) with a test compound;
- and
- c) assaying for the ability of the test compound to modulate the interaction of the prolines of the CD28 proliferation motif, which correspond to prolines 187 and 190 of the CD28 protein set forth in SEQ ID NO:2, with a molecule with which the CD28 proliferation motif interacts;
- wherein a determination that the test compound modulates the interaction of the prolines of the CD28 proliferation motif with a molecule with which the CD28 proliferation motif interacts, indicates that the test compound is a compound which selectively modulates T cell proliferation relative to T cell survival.
A cell-free method for identifying a compound which selectively modulates T cell proliferation relative to T cell survival, comprising;
- a) providing a polypeptide comprising a CD28 proliferation motif, in the context of a cell-free assay system;
- b) contacting the polypeptide of a) with a test compound;
- and
- c) assaying for the ability of the test compound to modulate the interaction of the prolines of the CD28 proliferation motif which correspond to prolines 187 and 190 of the CD28 protein set forth in SEQ ID NO:2, with a molecule with which the CD28 proliferation motif interacts;
- wherein a determination that the test compound modulates the interaction of the prolines of the CD28 proliferation motif with a molecule with which the CD28 proliferation motif interacts, indicates that the test compound is a compound which selectively modulates T cell proliferation relative to T cell survival.
Dependencies are inferred from the claim text.