Document Detail


Differential expression of intracellular and extracellular CB(2) cannabinoid receptor protein by human peripheral blood leukocytes.
MedLine Citation:
PMID:  23299999     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
mRNA encoding for the CB(2) cannabinoid receptor is expressed by many subsets of human peripheral blood leukocytes (PBL), but little is known about the resulting protein expression and function. Employing clones from the A549 and 293T cell lines that were constructed to express both full-length human CB(2) and GFP, we developed a flow cytometry assay for characterizing CB(2) protein expression. A monoclonal antibody directed against human CB(2) selectively stained the surface of transduced but not parental cell lines. When cells were fixed and permeabilized, imaging flow cytometry identified large stores of intracellular protein. Total cellular staining for CB(2) corresponded closely with the level of GFP expression. When exposed to Δ(9)-tetrahydrocannabinol, CB(2)-expressing cells internalized cell surface CB(2) receptors in a time- and dose-dependent manner. Applying these approaches to human PBL, CB(2) protein was identified on the surface of human B cells but not on T cells or monocytes. In contrast, when PBL were fixed and permeabilized, intracellular CB(2) expression was readily detected in all three subsets by both conventional and imaging flow cytometry. Similar to the protein expression pattern observed in fixed and permeabilized PBL, purified B cells, T cells, and monocytes expressed relatively equal levels of CB(2) mRNA by quantitative real-time RT-PCR. Our findings confirm that human PBL express CB(2) protein but that its distribution is predominantly intracellular with only B cells expressing CB(2) protein at the extracellular membrane. The differential role of intracellular and extracellular CB(2) receptors in mediating ligand signaling and immune function remains to be determined.
Authors:
Julie T Castaneda; Airi Harui; Sylvia M Kiertscher; Jeffrey D Roth; Michael D Roth
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't     Date:  2013-01-10
Journal Detail:
Title:  Journal of neuroimmune pharmacology : the official journal of the Society on NeuroImmune Pharmacology     Volume:  8     ISSN:  1557-1904     ISO Abbreviation:  J Neuroimmune Pharmacol     Publication Date:  2013 Mar 
Date Detail:
Created Date:  2013-02-28     Completed Date:  2013-08-16     Revised Date:  2014-03-07    
Medline Journal Info:
Nlm Unique ID:  101256586     Medline TA:  J Neuroimmune Pharmacol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  323-32     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Antibodies, Monoclonal / pharmacology
B-Lymphocytes / metabolism
Cell Line
Dose-Response Relationship, Drug
Dronabinol / pharmacology
Extracellular Space / metabolism
Flow Cytometry
Humans
Intracellular Space / metabolism
Leukocytes / metabolism*
Monocytes / metabolism
Real-Time Polymerase Chain Reaction
Receptor, Cannabinoid, CB2 / biosynthesis*
T-Lymphocytes / metabolism
Grant Support
ID/Acronym/Agency:
AI-28697/AI/NIAID NIH HHS; CA-16042/CA/NCI NIH HHS; R01 DA003018/DA/NIDA NIH HHS; R01-DA03018/DA/NIDA NIH HHS; R21 DA021813/DA/NIDA NIH HHS; R21-DA021813/DA/NIDA NIH HHS
Chemical
Reg. No./Substance:
0/Antibodies, Monoclonal; 0/Receptor, Cannabinoid, CB2; 7J8897W37S/Dronabinol
Comments/Corrections

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