Evidence for the nucleus accumbens as a neural substrate of heroin-induced immune alterations.

Administration of opioid drugs such as heroin produces several immunosuppressive effects, including decreases in natural killer (NK) cell activity, lymphocyte proliferative responses, and nitric oxide production. Interestingly, opioids have been shown to alter many immune parameters indirectly by modulating the immunoregulatory actions of the central nervous system. Recently, it has been demonstrated that morphine inhibits NK cell activity through a neural pathway that requires the activation of dopamine D(1) receptors in the nucleus accumbens shell. The present study examined whether the nucleus accumbens also mediates the effects of heroin, a more commonly abused opioid, on several parameters of immune status in Lewis rats. The results showed that bilateral administration of the dopamine D(1) receptor antagonist R-(+)-7-chloro-8-hydroxy-3-methyl-1-phenyl-2,3,4,5-tetrahydro-1H-3-benzazepine hydrochloride (SCH-23390; 0.015 and 0.15 microg/side) into the nucleus accumbens shell blocked decreases in splenic NK activity produced by heroin (3 mg/kg s.c.) but did not attenuate the suppression of splenocyte proliferative responses to concanavalin-A or lipopolysaccharide (LPS). A subsequent experiment was performed to evaluate the effect of D(1) receptor antagonism on LPS-induced expression of inducible nitric-oxide synthase (iNOS) in vivo. These results showed that intra-accumbens SCH-23390 administration prevented heroin-induced reductions of iNOS mRNA expression in spleen, liver, and lung tissues and attenuated the suppression of nitric oxide levels in plasma. Collectively, these findings indicate that nucleus accumbens dopamine D(1) receptors are critically involved in heroin-induced immune alterations.

Neuroimmune mechanisms of opioid-mediated conditioned immunomodulation.

Morphine administration elicits pronounced effects on the immune system, including decreases in natural killer (NK) cell activity and lymphocyte mitogenic responsiveness. These immune alterations can become conditioned to environmental stimuli that predict morphine as a result of Pavlovian conditioning processes. Prior work in our laboratory has shown that acute morphine exposure produces dopamine-dependent reductions of NK cell activity that are mediated peripherally by neuropeptide Y Y1 receptors. The present study examined the involvement of dopamine D1 and neuropeptide Y Y1 receptors in the conditioned immunomodulatory effects of morphine. Rats received two conditioning sessions during which an injection of morphine was paired with a distinctive environment which served as the conditioned stimulus (CS). The results show that systemic administration of the D1 antagonist SCH-23390 prior to CS re-exposure prevented the conditioned suppression of splenic NK activity but did not alter conditioned decreases in mitogen-induced lymphocyte proliferation. Furthermore, bilateral microinjections of SCH-23390 directly into the nucleus accumbens shell fully blocked conditioned changes in NK activity. In a subsequent manipulation, subcutaneous injection of the Y1 receptor antagonist BIBP3226 prior to CS re-exposure was also shown to prevent conditioned effects on NK activity. Collectively, these findings provide evidence that the nucleus accumbens shell plays an important role in conditioned immunomodulation and further suggest that the conditioned and unconditioned immunomodulatory effects of opioids involve similar receptor mechanisms.

Neuropeptide Y Y1 receptors mediate morphine-induced reductions of natural killer cell activity.

Morphine suppresses a number of immune parameters, such as natural killer (NK) cell activity and lymphocyte proliferation, by acting through mu-opioid receptors in the central nervous system. Prior studies have implicated the sympathetic nervous system in mediating the immunomodulatory effects of acute morphine treatment. However, the peripheral mechanism whereby morphine inhibits NK cell activity is not fully understood. The aim of the present study was to investigate the role of the sympathetic transmitter neuropeptide Y (NPY) in mediating morphine-induced immune alterations. The results showed that administration of the selective NPY Y1 receptor antagonist BIBP3226 blocked morphine's effect on splenic NK activity but did not attenuate the suppression splenocyte proliferative responses to Con-A or LPS. Furthermore, intravenous NPY administration produced a dose-dependent inhibition of splenic NK activity but did not suppress lymphocyte proliferation. Recent studies from our laboratory have demonstrated that morphine modulates NK activity through a central mechanism that requires the activation of dopamine D1 receptors in the nucleus accumbens. Results from the present study showed that microinjection of the D1 receptor agonist SKF-38393 into the nucleus accumbens shell induced a suppression of NK activity that was reversed by BIBP3226. Collectively, these findings demonstrate that NPY Y1 receptors mediate morphine's suppressive effect on NK activity and further suggest that opioid-induced increases in nucleus accumbens D1 receptor activation inhibit splenic NK activity via NPY released from the sympathetic nervous system.

Suppression of natural killer cell activity by morphine is mediated by the nucleus accumbens shell.

Despite a wealth of data indicating that morphine modulates immune status by acting at mu-opioid receptors in the brain, there is little known about how the opioid system interacts with other neurotransmitter systems to modulate specific immune parameters. The aim of the present study was to investigate whether dopaminergic projections to the nucleus accumbens are involved in morphine-induced suppression of splenic natural killer (NK) cell activity. The results indicate that administration of the dopamine D1 antagonist SCH-23390 into the nucleus accumbens shell, but not core, blocked morphine's suppressive effect on NK activity in male Lewis rats. In support of these findings, the effect of morphine was also prevented by intra-accumbens microinfusions of the dopaminergic immunotoxin anti-DAT-saporin. Additionally, administration of the D1 agonist SKF-38393 into the nucleus accumbens shell produced reductions in splenic NK activity comparable to morphine, suggesting a critical role for D1 receptors in the modulation of NK activity. Collectively, these findings demonstrate that dopaminergic inputs to the nucleus accumbens are critically involved in opioid-induced immunosuppression and suggest that opioid-induced increases in D1 receptor activation may have adverse consequences on immune status.

Buprenorphine produces naltrexone reversible alterations of immune status.

Substantial evidence demonstrates that administration of high efficacy mu opioid agonists such as morphine modulate the immune response in a dose-dependent and pharmacologically specific manner, indicating functional interactions between the opioid and immune systems. In contrast to the well-characterized immunomodulatory effects of high efficacy mu opioids, little is known about how these effects generalize to other clinically employed opioids and agonists of varying degrees of mu opioid receptor stimulation. Buprenorphine is a mu opioid agonist of intermediate efficacy that is used clinically for pain management and has recently been approved for the treatment of opioid dependence. Recent evidence indicates pharmacological and mechanistic differences between buprenorphine and morphine. Therefore, the aim of the present study was to investigate whether buprenorphine also possesses immunomodulatory properties. The results demonstrate that buprenorphine dose-dependently suppresses splenic natural killer cell activity, lymphocyte proliferation and IFN-gamma production in rats in a naltrexone reversible manner, demonstrating pharmacological specificity of buprenorphine-induced immune alterations.

Morphine-induced alterations of immune status are blocked by the dopamine D2-like receptor agonist 7-OH-DPAT.

Morphine administration produces profound effects on the immune system, including reductions in natural killer cell activity, mitogen-induced lymphocyte proliferation, and cytokine production. Although it has been established that the activation of central nervous system (CNS) micro-opioid receptors by morphine induces immunomodulation, little is known about the neural mechanisms underlying such processes. Interestingly, it has been shown that the dopamine (DA) D2-like receptor agonist 7-hydroxy-N,N-di-n-propyl-2-aminotetralin (7-OH-DPAT) blocks the effect of morphine on a number of behaviors that are mediated by central dopamine pathways. The present study examined whether dopamine is involved in the immunomodulatory effects of morphine. In separate experiments, 7-OH-DPAT was administered either systemically (subcutaneous, s.c.) or centrally (intracerebroventricularly, i.c.v.) prior to morphine treatment in male Lewis rats. The results demonstrate that both systemic and central administration of 7-OH-DPAT attenuate the suppressive effect of morphine on several measures of immune status. Overall, these findings provide the first evidence that CNS dopaminergic mechanisms are directly involved in morphine-induced immunomodulation.

Cocaine increases inducible nitric oxide synthase expression in rats: effects of acute and binge administration.

The present studies assessed the effects of acute and binge cocaine administration on the in vivo production of inducible nitric oxide synthase in a rat model of endotoxemia. Inducible nitric oxide synthase is a key enzyme involved in host defense and inflammatory responses consequent to infection through its production of nitric oxide. Male Lewis rats received subcutaneous injections of saline or selected doses of cocaine (7.5-30 mg/kg) at the same time as a 50 microg/kg dose of lipopolysaccharide (LPS). Animals in the binge cocaine experiments received two additional injections of cocaine or saline at 2 and 4 h after the initial injections. All animals were sacrificed 6 h after initial drug administration and tissues harvested for determination of iNOS protein levels. Plasma nitrite/nitrate levels were also assayed to assess any treatment-related differences in the degradative by-products of nitric oxide metabolism. Western blot analyses of iNOS expression indicated that both acute and binge cocaine treatment resulted in significant increases in iNOS expression in all tissues assayed. Furthermore, acute and binge cocaine treatment also dose-dependently increased plasma nitrite levels. These data suggest that cocaine may impact resistance to gram-negative infections and severity of these infections via modulation of nitric oxide parameters.

Heroin-associated environmental stimuli modulate the expression of inducible nitric oxide synthase in the rat.

RATIONALE: Heroin (diacetylmorphine) administration has been shown to alter the induction of nitric oxide, a molecule known to play a critical role in the regulation of immune responses and resistance to infectious challenges. There is evidence that the physiological and behavioral effects of opioids can be conditioned to environmental stimuli associated with drug administration. OBJECTIVES: The present study tests whether environmental stimuli paired with the administration of heroin alter the expression of inducible nitric oxide synthase (iNOS), one of the isoforms of the enzyme responsible for nitric oxide production. METHODS: The experimental group of rats received three conditioning sessions each involving a subcutaneous injection of heroin (1.0 mg/kg) immediately upon being placed in a chamber that served as the conditioned stimulus. Following the conditioning sessions, the rats remained in their home cages for 6 days to recover from the effects of heroin per se. The rats then were re-exposed to the conditioning chamber in the absence of heroin and immediately following the session were injected subcutaneously with lipopolysaccharide (LPS) to induce iNOS expression. Six hours later, the expression of iNOS messenger ribonucleic acid (mRNA) and protein was determined in spleen, lung, and liver using real-time reverse transcriptase polymerase chain reaction (RT-PCR) and western blotting, respectively. Control procedures were used to determine whether the alterations in iNOS expression were due to conditioning processes. RESULTS: The results showed that exposure to the conditioned stimulus produced a pronounced reduction in the expression of iNOS mRNA and protein in spleen, lung, and liver tissue. Control procedures showed that this effect was due to conditioning processes. CONCLUSIONS: This study provides the first evidence that heroin-induced alteration of iNOS expression can be conditioned to environmental stimuli, and suggest that environmental stimuli associated with drug use may contribute to the alteration in susceptibility to infection.

Self-administration of heroin produces alterations in the expression of inducible nitric oxide synthase.

Nitric oxide plays a critical role in the immune response, and our studies have shown that heroin induces a reduction in the expression of iNOS, the enzyme responsible for nitric oxide production. The present study evaluated the effect of heroin self-administration on iNOS expression using a three-group design. Group one (self-administration) was trained to press a lever for i.v. administration of heroin. Group two (yoked heroin) received a simultaneous equivalent infusion of heroin determined by the responses of a 'partner' animal in the first group. A third group (yoked saline) also was yoked to the first group, but received i.v. injections of saline. Immediately following the last session, all rats received an injection of lipopolysaccharide (LPS) to induce iNOS expression. About 6 h after the injection of LPS, iNOS mRNA and protein expression were determined in spleen, lung, and liver. Additionally, the accumulation of plasma nitrite/nitrate, the more stable end products of nitric oxide degradation were measured. Although there was not a consistent difference between the self-administering and yoked-heroin animals, the results show that rats will self-administer a sufficient amount of heroin to induce a pronounced, widespread reduction in the expression of iNOS.

An examination of nucleus accumbens cell firing during extinction and reinstatement of water reinforcement behavior in rats.

Electrophysiological recording procedures were used to examine nucleus accumbens (Acb) cell firing in rats (n = 13) during water reinforcement sessions consisting of three phases. During phase one (maintenance), a lever press resulted in water reinforcement (fixed ratio 1; 0.05 ml/press) paired with an auditory stimulus (0.5 s). Of 128 Acb neurons recorded during maintenance, 40 cells (31%) exhibited one of three types of neuronal firing patterns described previously [J. Neurosci. 14 (12) (1994) 7735-7746; J. Neurosci. 20 (11) (2000) 4255-4266]. Briefly, Acb neurons exhibited increases in firing rate within seconds preceding the reinforced response (type PR) or increases (type RFe) or decreases (type RFi) in activity seconds following response completion. In phase two (extinction), subsequent lever pressing had no programmed consequences (i.e., water reinforcement and the auditory stimulus were not presented). After 30 min of no responding, animals were given water reinforcement/auditory stimulus 'primes' to reestablish lever pressing behavior during the third phase (reinstatement). Results indicated that all types of phasic neurons (PR, RFe and RFi) exhibited an attenuated firing rate during extinction, and in some cases recovery of patterned discharges were observed during reinstatement. No significant changes in cell firing were observed for any cell type during presentation of the stimulus prime used to reestablish operant responding following extinction. These findings are discussed in terms of how Acb neurons process information related to 'natural' reinforcers versus drugs of abuse.