Selective effect of methoxyindoles on the lymphocyte proliferation and melatonin binding to activated human lymphoid cells.
Three pineal methoxyindoles (melatonin (Mel), 5-methoxytryptamine (5-MTA) and 5-methoxytryptophol (5-MTO)) were studied for their ability to influence the proliferative response of human peripheral blood lymphocytes (PBL) and tonsillar lymphocytes (TL) following activation with concanavalin A (ConA) in vitro. The ConA-stimulated DNA synthesis was affected in a different dose-dependent mode by the methoxyindoles tested. Melatonin and 5-MTO inhibited and 5-MTA increased the ConA-induced [3H]thymidine incorporation in PBL and TL. The initial screening for 2-[125I]iodomelatonin binding using a single point assay revealed significantly increased specific binding to PBL and TL after 72-h stimulation with ConA as compared to the non-activated cell cultures. Coincubation of separate lymphocyte cultures with ConA and Mel or 5-MTO resulted in inhibition of the specific 2-[125I]iodomelatonin binding (85% and 74%, respectively). The specific binding determined in the presence of 5-MTA did not differ from control values. Series of saturation and competition experiments were performed to examine the binding characteristics of ConA-stimulated lymphocytes for 2-[125I]iodomelatonin. The radioligand labelled binding sites of high affinity (Kd = 0.14 +/- 0.03 nM) and low capacity (Bmax = 6.8 +/- 1.5 fM/mg protein). Competitive studies with a variety of indoles determined the following order of relative potency for inhibition of 2-[125I]iodomelatonin binding in TL: 2-iodomelatonin > melatonin > > 5-methoxytryptophol. 5-Methoxytryptamine did not show displacement potency for the labelled ligand. Collectively, our data suggest that pineal hormones might be directly involved in the regulation of the T-lymphoproliferative response of human lymphoid cells. We show the availability of melatonin receptors, which seem to be an intrinsic characteristic of activated human lymphocyte populations. While the effects of Mel and 5-MTO can be linked to the binding sites described, it is unlikely that serotonin agonists like 5-MTA may act through the same sites to influence the mitogen-stimulated lymphocyte proliferation.
Department of Immunoneuroendocrinology, Bulgarian Academy of Sciences, Sofia, Bulgaria.
R
R
Konakchieva
S
S
Kyurkchiev
I
I
Kehayov
P
P
Taushanova
L
L
Kanchev
D008735:5-Methoxytryptamine / Q000276:immunology*; D000328:Adult; D002455:Cell Division / Q000276:immunology; D002478:Cells, Cultured / Q000276:immunology; D003208:Concanavalin A / Q000494:pharmacology; D004306:Dose-Response Relationship, Immunologic; D006801:Humans; D007211:Indoles / Q000276:immunology*; D008213:Lymphocyte Activation / Q000276:immunology*; D008214:Lymphocytes / Q000166:cytology* / Q000378:metabolism; D008550:Melatonin / Q000276:immunology* / Q000378:metabolism; D008875:Middle Aged; D008934:Mitogens / Q000494:pharmacology; D011485:Protein Binding / Q000276:immunology; D013205:Staphylococcal Protein A / Q000494:pharmacology; D013745:Tetanus Toxoid / Q000494:pharmacology
D007211:Indoles; D008934:Mitogens; D013205:Staphylococcal Protein A; D013745:Tetanus Toxoid; D003208:Concanavalin A; D008735:5-Methoxytryptamine; C002465:methoxytryptophol; D008550:Melatonin