Ca2+-independent protein kinase Cs mediate heterologous desensitization of leukocyte chemokine receptors by opioid receptors.
Heterologous desensitization of chemokine receptors by opioids has been considered to contribute to their immunosuppressive effects. Previous studies show that Met-enkephalin, an endogenous opioid, down-regulates chemotaxis of selected chemokine receptors via phosphorylation. In the present study, we further investigated the molecular mechanism of such cross-regulation. Our data showed that preincubation with Met-enkephalin inhibited both MIP-1 alpha-mediated chemotaxis and Ca(2+) flux of monocytes in a dose-dependent manner. The inhibitory effects were maximal using nanomolar concentrations of activating chemokines, a concentration found in physiological conditions. A decrease both in chemokine receptor affinity and in coupling efficiency between receptors and G protein were observed, which directly contributed to the desensitization effects. However, comparing with chemokines such as MIP-1 alpha and MCP-1, opioids did not elicit a calcium flux, failed to induce MIP-1 alpha receptors internalization, and mediated a less potent heterologous desensitization. We hypothesized that these differences might originate from the involvement of different protein kinase C (PKC) isotypes. In our studies, opioid-mediated down-regulation of MIP-1 alpha receptors could be blocked by the general PKC inhibitor calphostin C, but not by the calcium-dependent classic PKC inhibitor Go6976. Western blotting analysis and immunofluorescent staining further showed that only calcium-independent PKCs were activated upon opioid stimulation. Thus, opioids achieve desensitization of chemokine receptors via a unique pathway, involving only calcium-independent PKC isotypes.
Zhang
Ning
N
Laboratory of Molecular Immunoregulation, Intramural Research Support Program, Frederick, Maryland 21702-1201, USA.
Hodge
Dave
D
Rogers
Thomas J
TJ
Oppenheim
Joost J
JJ
D002118:Calcium / Q000494:pharmacology; D002478:Cells, Cultured; D054407:Chemokine CCL4; D002634:Chemotaxis, Leukocyte / Q000187:drug effects / Q000276:immunology / Q000502:physiology*; D004744:Enkephalin, Methionine / Q000494:pharmacology*; D006706:Homeostasis; D006801:Humans; D007962:Leukocytes / Q000276:immunology*; D019402:Macrophage Inflammatory Proteins / Q000502:physiology; D009000:Monocytes / Q000187:drug effects / Q000276:immunology / Q000502:physiology*; D011493:Protein Kinase C / Q000097:blood*; D019707:Receptors, Chemokine / Q000097:blood*; D011957:Receptors, Opioid / Q000502:physiology*
D054407:Chemokine CCL4; D019402:Macrophage Inflammatory Proteins; D019707:Receptors, Chemokine; D011957:Receptors, Opioid; D004744:Enkephalin, Methionine; D011493:Protein Kinase C; D002118:Calcium