The effects of β‐adrenoceptor stimulation on adhesion of human natural killer cells to cultured endothelium

Abstract

1. The circulation of natural killer (NK) cells in vivo is influenced by physical exercise, mental stress, and infusion of beta-adrenoceptor agonists. We have previously presented in vitro data, showing that beta 2-adrenoceptor agonists induce detachment of NK cells from endothelial cells (EC), supporting the hypothesis that NK cells can be recruited from the marginating pool in blood vessels. 2. Because NK cells as well as EC express beta 2-adrenoceptors, the present study was conducted to investigate whether stimulation of the beta-adrenoceptors on NK cells, EC or both cell types is required to induce detachment from EC. 3. Cells were pretreated (15 min) with a selective beta 2-adrenoceptor antagonist, GR81706, at various concentrations. The duration of beta-adrenoceptor blockade was tested by determining the adenosine 3',5'-cyclic monophosphate (cyclic AMP) production induced by terbutaline (a beta 2-adrenoceptor specific agonist). This receptor-mediated response was effectively inhibited for at least 4 h, whereas the cyclic AMP production in response to forskolin (a direct activator of adenylate-cyclase) was not affected. 4. Functional adhesion assays were then performed to determine the role of beta-adrenoceptors on the different cell types involved (NK and EC) in catecholamine-induced detachment. Peripheral blood mononuclear cells were allowed to adhere for 1 h to monolayers of unstimulated EC in the presence or absence of cyclic AMP inducing agents, and the percentage of NK cells in the adhering lymphocyte fraction was determined by flow cytometry. 5. Both adrenaline (10(-5) M) and forskolin (10(-5) M) caused detachment of NK cells from EC. After blockade of the P2-adrenoceptors on NK cells by pretreatment with GR81706 (10-6 M), the effect of adrenaline on NK cells adhesion was pretented; after blockade of the beta2-adrenoceptors on EC, NK cell adhesion was still significantly reduced by adrenaline. In all cases, forskolin caused detachment of NKcells.6. To establish further that stimulation of beta-adrenoceptors on NK cells is sufficient to cause detachment,we showed that adrenaline also reduced adhesion of NK cells to monolayers of Chinese hamster ovary cells, which do not express beta-adrenoceptors.7. Together, these results show that stimulation of beta2-adrenoceptors on NK cells negatively influences their capacity to adhere to EC, and that beta2-adrenoceptors on EC play a negligible role in this phenomenon.