Effects of endogenous and synthetic opioid peptides on neutrophil function in vitro.

BACKGROUND: Opioid peptides released from immunocytes during inflammation and stress in critically ill patients are associated with an altered immune response. Moreover, concentrations of opioid peptides are increased in peripheral blood and at the sites of inflammatory reactions. METHODS: Using flow cytometric assay of whole human blood, we investigated direct effects of endogenous and synthetic opioid peptides on surface expression of complement receptors CD35 and CD11b/CD18 and Fcã receptor III CD16, and superoxide anion generation of neutrophils. RESULTS: The endogenous opioid peptides beta-endorphin(1-31) and met-enkephalin, representing the N-terminal fragment of beta-endorphin(1-31), and the synthetic delta opioid receptor agonists D-Ala(2)-D-Leu(5)-enkephalin and D-Pen(2)-enkephalin produced concentration-dependent stimulation of neutrophil activity. Incubation with met-enkephalin 10(-7) M or beta-endorphin(1-31) 10(-7) M led to an increase in receptor expression of up to 10% (met-enkephalin) and 15% (beta-endorphin(1-31)). After incubation with D-Ala(2)-D-Leu(5)-enkephalin or D-Pen(2/5)-enkephalin, receptor expression was increased by up to 30%. This correlated with concentration-dependent stimulation of the production of reactive oxygen intermediates, as shown by an increase of up to 40% in oxidative burst activity. All effects were abolished after preincubation with naloxone or with the selective delta opioid antagonist naltrindole, whereas the selective micro receptor antagonist d-Phe-Cys-Tyr-D-Trp-Orn-Thr-Pen-Thr-NH(2) showed only partial inhibitory effects. CONCLUSIONS: Our data suggest a delta opioid receptor-mediated stimulatory effect on neutrophil function. beta-Endorphin(27-31), the C-terminal fragment of beta-endorphin(1-31), did not alter neutrophil function, indicating that beta-endorphin(1-31) mediates its effect on neutrophils via the N-terminal fragment. This study may contribute to a better understanding of neuroimmune interaction.

Subcellular distribution of delta-opioid receptors in the rat spinal cord: an approach using a three-dimensional reconstruction of confocal series of immunolabelled neurons.

Using a specific rat monoclonal anti-idiotypic antibody we have examined the subcellular distribution of -opioid receptors in various neuronal subtypes of the rat spinal cord. The immunofluorescence was detected with a confocal microscope and in some cases serial images were processed for a three-dimensional (3-D) reconstruction of the neurons. Immunolabelling was found to be distributed throughout the spinal cord grey matter specially in the most superficial layers of the dorsal horn, around the central canal and in the region of motoneurons of the ventral horn. The 3-D reconstruction made on large neurons of lamina IX in the ventral horn and on neurons of lamina X around the central canal allowed the visualization of 5 -opioid receptors in the cytoplasm of the soma and proximal neurites of immunofluorescent neurons. Some immunolabelled receptors were also detected at the level of the plasma membrane of the cell bodies and in the nuclear matrix. Interestingly, a particular arrangement of delta-opioid receptors organized along parallel alignments was observed on the plasma membrane of some neurons. This study emphasizes the potential usefulness of a 3-D reconstruction in the study of the spatial arrangement of cellular components.