Characterisation of nicotine receptors on human peripheral blood mononuclear cells (PBMC).
OBJECTIVE
The aim of the work was to characterise the nAChRs on human PBMC.


METHODS
PBMC were isolated from human blood buffy coats provided by the blood transfusion service and were used for radioligand binding studies with [(3)H]-nicotine. RT-PCR experiments were used to determine nAChR subunit expression while immunoblotting experiments were used to confirm that nAChR subunits identified by RT-PCR were translated into protein.


RESULTS
Binding studies suggested the presence of one binding site for (-)- nicotine on human peripheral blood lymphocytes. Competition studies showed that only (-)- nicotine, epibatidine and alpha-bungarotoxin, displaced radiolabelled nicotine from cells. RT-PCR studies demonstrated mRNA for alpha4, alpha5, alpha7, beta1 and beta2 nAChRs subunits in PBMC. Expression of mRNA for the a5 subunit of nAChR was observed in all lymphocyte samples tested. In contrast, the expression pattern of mRNAs for alpha4, alpha7, beta1, and beta2 mRNAs subunits of nAChRs, varied between samples. Western blot analysis showed that protein for alpha4, alpha5, and alpha7 and beta2 nAChR subunits was expressed in most, but not all of the PBMC samples tested but some of the bands obtained were faint.


CONCLUSIONS
The results obtained suggest that human PBMC contain nAChRs containing alpha4beta2, alpha4beta2alpha5, and/or alpha7 subunits.
Wongsriraksa
A
A
Department of Life Sciences, University of Hertfordshire, Hertfordshire, UK.
Parsons
M E
ME
Whelan
C J
CJ
D000818:Animals; D002478:Cells, Cultured; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000378:metabolism*; D009538:Nicotine / Q000378:metabolism; D018722:Nicotinic Agonists / Q000378:metabolism; D021122:Protein Subunits / Q000378:metabolism*; D011869:Radioligand Assay; D011978:Receptors, Nicotinic / Q000378:metabolism*
D018722:Nicotinic Agonists; D021122:Protein Subunits; D011978:Receptors, Nicotinic; D009538:Nicotine