Cannabinoid 2 (CB
Cannabinoid receptor 2 (CB 2 ) is expressed in B lymphocytes and is involved in immune regulation. Mouse splenic B cells express three CB 2 transcripts utilizing two different first exons. Human peripheral blood B cells express one CB 2 transcript utilizing one first exon. Alignment of sequenced RACE products to either the mouse or human genome reveals that isolated transcripts contain previously unidentified transcriptional start sites (TSS). B cells from mouse and human preferentially express one transcript, exon 1a in mouse and exon 1 in human. Multiple CB 2 TSSs are utilized in mouse splenic B cells and one TSS in human peripheral blood B cells. The defining of the receptor gene TSSs in these cells provides materials and methods for therapeutically regulating immune function, including antibody isotype switching, using compounds such as inhibitory nucleic acids that down-regulate expression of the B cell CB 2 gene (e.g., RNAi molecules).
1. A method of screening for an allergen or anti-allergen, comprising:
providing a cell having a construct comprising a B cell cannabinoid receptor 2 gene promoter and a reporter gene operably linked thereto;
contacting the cell with one or more compounds to be tested as an allergen or anti-allergen; and
measuring the resulting reporter gene expression.
2. The method of claim 1 , wherein the CB 2 receptor gene promoter is a human CB 2 receptor gene promoter sequence.
3. The method of claim 1 , wherein the CB 2 receptor gene promoter is a mouse CB 2 receptor gene promoter sequence.
4. The method of claim 1 , wherein the CB 2 receptor gene promoter is a human CB 2 receptor gene promoter sequence comprising a nucleic acid sequence selected from among SEQ ID NO:6, SEQ ID NO:115, SEQ ID NO:116, SEQ ID NO:117, SEQ ID NO:135, SEQ ID NO:136; SEQ ID NO:174, or SEQ ID NO:175.
5. The method of claim 1 , wherein the CB 2 receptor gene promoter is a mouse CB 2 receptor gene promoter sequence comprising a nucleic acid sequence selected from among SEQ ID NO:1, SEQ ID NO:111, SEQ ID NO:112, SEQ ID NO:113, or SEQ ID NO:114.
6. The method of claim 1 , wherein the construct comprises the transcription start site associated with the B cell CB 2 receptor gene promoter region.
7. The method of claim 1 , wherein the construct comprises at least one region selected from among the transcription start site (tss) for human exon 1, or at least a portion of the 5′ untranslated region (UTR) for human exon 1.
8. The method of claim 1 , wherein the construct comprises at least one region selected from among the transcription start site (tss) for mouse exon 1a, mouse exon 1b, or mouse exon 2, or at least a portion of the 5′ UTR for mouse exon 1a, mouse exon 1b, or mouse exon 2.
9. The method of claim 1 , wherein the cell is a eukaryotic cell.
10. The method of claim 1 , wherein the cell is a non-human mammal cell.
11. The method of claim 1 , wherein the cell is a human cell.
12. The method of claim 1 , wherein the cell has been genetically modified to produce the CB 2 receptor and did not have the capability in the absence of the genetic modification.
13. The method of claim 1 , wherein the cell has been genetically modified to increase the amount of CB 2 endogenously produced by the cell.
14. The method of claim 1 , wherein the cell is a B-cell.
15. The method of claim 1 , wherein the cell is a non-B-cell.
16. The method of claim 1 , wherein said contacting comprises contacting the cell with an allergen and one or more compounds to be tested as an anti-allergen; and wherein said method further comprises comparing the resulting reporter gene expression to the reporter gene expression of a cell contacted with only an allergen.