Determining the molecular basis of opiate and opioid modulation of immune function is critical to understanding such apparently diverse phenomena as the immunosuppressive effects of drug abuse and stress, their correlations with progression of HIV infection to clinical AIDS, and the possibilities for therapeutic intervention. T cell functions are critical to both cellular and humoral immunity. The immunodeficiency of AIDS is associated with specific declines in CD4+ T cell dependent functions and impaired T cell receptor signal transduction. CD8+ T cell derived factors suppress HIV replication. There is evidence that the opioid peptide met-enkephalin ameliorates HIV associated T cell defects in vivo. Although some opiate immunomodulatory effects may be mediated through the CNS there is abundant evidence for immunomodulation effected by direct opioid immunocyte interactions. And, cells of the immune system express opioid receptors thought to be similar to those in neural tissue. The objective of this proposal is to identify the opioid receptors responsible for the apparently opposite effects of methionine enkephalin in vitro on activation of purified CD4 (helper type) and CD8 (suppressor type) positive T cells and to initiate an investigation of the underlying biochemical mechanisms. Immobilized monoclonal antibody against the T cell receptor associated CD3 complex (anti-CD3 mAb) elicits specific polyclonal activation of T cells in the absence of accessory cells. The cascade of signal transduction, intracellular second messengers and induction of specific gene transcription elicited by anti- CD3 mAb is well characterized and essentially identical to that elicited by physiologic T cell activation. This model will be used to identify specific receptors in CD4+ and CD8+ T cells through which opioid peptide modulation of T cell activation is mediated, and should provide an excellent system in which to identify the specific T cell activation event(s) affected. In the first aim, RT-PCR will be used to determine if delta and mu receptor messenger RNAs identical or closely related to those recently determined in neural tissues are expressed in CD4+ and CD8+ T cells purified from murine spleen. Although Met-enkephalin is primarily considered a delta opioid receptor agonist there is evidence to suggest that both delta and mu opioid receptors may be involved in its effects on T cells. In the second aim, radioreceptor assays will be used to quantify and characterize the putative delta and mu opioid receptor binding sites in CD4+ and CD8+ T cells, and agonist and antagonist binding to these sites will be correlated with modulation of anti-CD3 stimulated T cell proliferation. The results of Aim 2 will confirm that expression of appropriate binding sites parallels expression of specific opioid receptor messenger RNA and may be informative as to whether the ligand specificity of expressed receptors is dependent in part on cell background.
The third aim i s to identify the early signal transduction events of T cell activation which are postulated to be modulated by activity of delta and mu opioid receptors and result in the observed opioid peptide effects on T cell proliferation. Opioid peptide effects on anti-CD3 mAb stimulated phospholipase C dependent phosphoinositide hydrolysis, specific tyrosine kinase phosphorylation, and elevation of intracellular free calcium will be investigated.

Agency
National Institute of Health (NIH)
Institute
National Institute on Drug Abuse (NIDA)
Type
Research Project (R01)
Project #
1R01DA008801-01A1
Application #
2121547
Study Section
Sociobehavioral Subcommittee (DAAR)
Project Start
1994-09-01
Project End
1997-07-31
Budget Start
1994-09-01
Budget End
1995-07-31
Support Year
1
Fiscal Year
1994
Total Cost
Indirect Cost
Name
University of Texas Sw Medical Center Dallas
Department
Biochemistry
Type
Schools of Medicine
DUNS #
City
Dallas
State
TX
Country
United States
Zip Code
75390
Shelton, Keith L; Beardsley, Patrick M (2008) Effect of drug-paired exteroceptive stimulus presentations on methamphetamine reinstatement in rats. Pharmacol Biochem Behav 90:434-40
Beardsley, Patrick M; Howard, James L; Shelton, Keith L et al. (2005) Differential effects of the novel kappa opioid receptor antagonist, JDTic, on reinstatement of cocaine-seeking induced by footshock stressors vs cocaine primes and its antidepressant-like effects in rats. Psychopharmacology (Berl) 183:118-26
Abougergi, Marwan S; Gidner, Sarah J; Spady, David K et al. (2005) Fas and TNFR1, but not cytolytic granule-dependent mechanisms, mediate clearance of murine liver adenoviral infection. Hepatology 41:97-105
Shelton, Keith L; Hendrick, Elizabeth; Beardsley, Patrick M (2004) Interaction of noncontingent cocaine and contingent drug-paired stimuli on cocaine reinstatement. Eur J Pharmacol 497:35-40
Miller, Bonnie C; Eckman, Elizabeth A; Sambamurti, Kumar et al. (2003) Amyloid-beta peptide levels in brain are inversely correlated with insulysin activity levels in vivo. Proc Natl Acad Sci U S A 100:6221-6
Nguyen, Khanh; Miller, Bonnie C (2002) CD28 costimulation induces delta opioid receptor expression during anti-CD3 activation of T cells. J Immunol 168:4440-5
Miller, B (1996) delta opioid receptor expression is induced by concanavalin A in CD4+ T cells. J Immunol 157:5324-8
Miller, B C; Thiele, D; Hersh, L B et al. (1996) A secreted peptidase involved in T cell beta-endorphin metabolism. Immunopharmacology 31:151-61
Safavi, A; Miller, B C; Cottam, L et al. (1996) Identification of gamma-endorphin-generating enzyme as insulin-degrading enzyme. Biochemistry 35:14318-25