Methods of screening for B cell activity modulators
View Patent ↗The invention provides for the identification of all genes, whether known or novel, which are differentially expressed within and among B cells, making possible the characterization of their temporal regulation and function in the B cell response and/or in B cell mediated disorders. Expression profiles, nucleic acids and proteins are provided for differing states of B cells, including resting, naive, activated, tolerant and immunosuppressed B cells. The present invention makes possible the identification and characterization of targets useful in prognosis, diagnosis, monitoring, rational drug design, and/or therapeutic intervention of immune system disorders.
1. A method of screening drug candidates, comprising:
a) providing a B cell that expresses one or more expression profile genes selected from the group consisting of carb anh II, IgD, CD72, SATB1, ApoE, CD83, cyclin D2, Cctq, MEF-2C, TGIF, Aeg-2, 1ck, E2-20K, pcp-4, kappa V, neurogranin, NAB2, gfi-1 hIP-30, TRAP, bmk, CD36, Evi-2, vimetin, Ly6E.1 and c-fes;
b) adding a drug candidate to the B cell;
c) determining the expression level of at least one gene of the expression profile genes in the B cell;
d) comparing the expression level of the at least one gene in the B cell with the expression level of the at least one gene in a control cell not contacted with the drug candidate to determine (i) whether expression of carb anh II, CD72, SATB1, ApoE, CD83, cyclin D2, Cctg, MEF-2C, TGIF, Aeg-2, 1ck, E2-20K, pcp-4, kappa V, neurogranin, NAB2 and/or gfi-1 is increased in the B cell relative to a control cell not contacted with the drug candidate, or (ii) whether expression of Ly6E.1, vimentin. hIP-30, TRAP, bmk, CD36, Evi-2 and/or c-fes is decreased in the B cell relative to the control cell; and
e) identifying the drug candidate as a potential modulator of B cell tolerance if the expression level of a gene listed in (i) is increased and/or the expression level of a gene listed in (ii) is decreased.
2. The method according to claim 1 , further comprising performing a binding assay to determine if the drug candidate identified in step e) binds to the protein encoded by the at least one gene.
3. The method according to claim 1 , further comprising performing an assay to determine if the drug candidate identified in step e) modulates an activity of the protein encoded by the at least one gene.
4. The method according to claim 1 , wherein the expression levels of a plurality of expression profile genes are determined and compared.
5. The method according to claim 4 , wherein the expression levels of at least three expression profile genes are determined and compared.
6. The method according to claim 5 , wherein the expression levels of at least five expression profile genes are determined and compared.
7. The method of claim 1 , wherein the expression level is determined from the amount of transcript expressed by the at least one gene.
8. The method of claim 1 , wherein the expression level is determined from the amount of protein expressed by the at least one gene.