The suppressive effect of 1.0 mg/kg buprenorphine on IFN- production is attenuated by administration of the opioid receptor antagonist, naltrexone.K.A. Carrigan et al., Int Immunopharmacol 2004, 4:419-428

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CD4+ T lymphocytes are the primary cell target for human immunodeficiency virus-1 (HIV-1), and these cells are known to express opioid receptors.

Due to the need for new treatment approaches to HIV-1 infection, we sought to determine whether the non-selective opioid receptor antagonist naltrexone would affect HIV-1 expression in CD4+ lymphocyte cultures and whether naltrexone would alter the antiviral properties of zidovudine (AZT) or indinavir.

Activated CD4+ lymphocytes were infected with a monocytotropic or T-cell tropic HIV-1 isolate, and p24 antigen levels were measured in supernatants of drug-treated or untreated (control) cultures.

While naltrexone alone did not affect HIV-1 expression, at a concentration of 10−12–10−10 M naltrexone increased the antiviral activity of AZT and indinavir 2–3-fold.

Similar findings with a -opioid receptor (KOR) selective antagonist supported the possible involvement of KOR in naltrexone’s potentiation of the antiretroviral drugs.

The results of this in vitro study suggest that treatment of alcohol or opiate dependent HIV-1-infected patients with naltrexone is unlikely to interfere with the activity of antiretroviral drugs. Also, based upon naltrexone’s safety profile and its synergistic activity in vitro, these findings suggest clinical trials should be considered of naltrexone as an adjunctive therapy of HIV-1 infection.

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The analgesic property of opiates has been known since ancient times. Only recently has an appreciation of the broad effects of opioids on the inflammatory response emerged. Acting largely through -, - and -opioid G protein-coupled receptors on T lymphocytes and macrophages, cognate ligands modulate many activities of these cells, including cytokine production. In addition to acting as chemotactic stimuli, opioids can, through the process of heterologous cross-desensitization, act as stop signals in leukocyte trafficking. When administered into the central nervous system, certain chemokines can cross-desensitize to the analgesic effect of opioids. We propose that opioids should be considered members of the cytokine family and that future research on opioids could yield new therapies for inflammatory and infectious diseases, including HIV-1 infection.

“We propose that opioids should be considered members of the cytokine family and that future research on opioids could yield new therapies for inflammatory and infectious diseases, including HIV-1 infection.”

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Opioid and chemokine receptor expression by T cells, monocytes, microglial cells and neurons. Neuron–microglial cell communication occurs through the release of various chemical mediators, including the production of CX3CL1 (fractalkine) by neurons and the production of additional chemokines, such as CCL3, CCL4 and CCL5, by microglial cells.

The chemokines produced by microglial cells (and astrocytes) in the brain act as chemoattractants and/or activators of circulating T cells and monocytes. The production of CCL2 is a potent chemoattractant for the passage of monocytes through the blood–brain barrier. In addition, endogenous opioids, produced in the brain and by cells of the immune system, act to regulate circulating immune cells and microglial-cell inflammatory activities.

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-opioid receptor (MOR) ligands also signal an upregulation in the expression of the major HIV coreceptors CCR5 and CXCR4. By contrast, -opioid receptor (KOR) activation directly (and probably indirectly) inhibits the expression of proinflammatory cytokines and chemokines. This results in a depressed state of cellular activation and reduced HIV replication. In addition, KOR signals an inhibition of the expression of CCR5 and CXCR4, leading to reduced HIV binding and reduced chemotaxis of potentially susceptible target cells.

Model describing the influence of opioids on HIV replication. MOR activation of macrophages and potentially T cells and microglia initiates an increase in the expression of chemokines CCL2, CCL5 and CXCL10, which might act to attract susceptible T cells, monocytes and/or macrophages or microglia to the site of HIV infection.

A substantial proportion of HIV-1-infected individuals are intravenous drug users (IVDUs) who abuse opiates. Opioids induce a number of immunomodulatory effects that may directly influence HIV-1 disease progression. In the present report, we have investigated the effect of opioids on the expression of the major HIV-1 coreceptors CXCR4 and CCR5. For these studies we have focused on opiates which are ligands for the -opioid receptor. Our results show that DAMGO, a selective -opioid agonist, increases CXCR4 and CCR5 expression in both CD3+ lymphoblasts and CD14+ monocytes three- to fivefold. Furthermore, DAMGO-induced elevation of HIV-1 coreceptor expression translates into enhanced replication of both X4 and R5 viral strains of HIV-1. We have confirmed the role of the –opioid receptor based on the ability of a -opioid receptor-selective antagonist to block the effects of DAMGO. We have also found that morphine enhances CXCR4 and CCR5 expression and subsequently increases both X4 and R5 HIV-1 infection. We suggest that the capacity of -opioids to increase HIV-1 coreceptor expression and replication may promote viral binding, trafficking of HIV-1-infected cells, and enhanced disease progression.

“Our results show that DAMGO, a selective -opioid agonist, increases CXCR4 and CCR5 expression in both CD3+ lymphoblasts and CD14+ monocytes three- to fivefold. Furthermore, DAMGO-induced elevation of HIV-1 coreceptor expression translates into enhanced replication of both X4 and R5 viral strains of HIV-1.”

-Opioid modulation of HIV-1 coreceptor expression and HIV-1 replication Steele A.D, et al., 309:99–10 (2003)

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The effect of month-long naltrexone (NTX) treatment at a daily oral dose of 0.5 mg/kg/day was contrasted with placebo (PLC) in a double-blind study with conjoint clinical and biochemical evaluations of therapeutic effects. Modest clinical benefits were achieved with both PLC and NTX, with marginally better overall results following NTX, and degree of improvement appeared to be related to plasma chemical profiles. Massively elevated levels of -endorphin were observed in all children with assays using C-terminal antibody but not with an N-terminal antibody assay. In addition, 70% of the children exhibited abnormally low levels of adrenocorticotropic hormone, and smaller subsets exhibited elevated norepinephrine (6@%), arginine-vasopressin (SO%), and serotonin (20%). The best clinical responders exhibited the clearest normalization of the elevated plasma chemistries, especially in C-terminal-&endorphin and serotonin. There was some evidence of therapeutic carry-over effects in both clinical and biochemical measures in those children who received NTX before PLC. The results suggest that NTX only benefits a subgroup of autistic children, who may be identified by the presence of certain plasma abnormalities. These results suggest a possible linkage between abnormal plasma chemistries, especially those related to the pro-opiomelanocortin system, and autistic symptoms.

Low-dose naltrexone effects on plasma chemistries and clinical symptoms in autism: a double-blind, placebo-controlled studyBouvard M.P., et al.

“The results suggest that NTX only benefits a subgroup of autistic children, who may be identified by the presence of certain plasma abnormalities. These results suggest a possible linkage between abnormal plasma chemistries, especially those related to the pro-opiomelanocortin system, and autistic symptoms.”

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Low-Dose Naltrexone Therapy Improves Active Crohn’s Disease

Smith J.P., et al., 102:820–828, 2007

STUDY HIGHLIGHTSWhat Is Current KnowledgeThe current medical therapy of Crohn’s disease includes medications

that target the immune system or inflammatory modulators. Opioid systems (peptides and receptors) play an integral role in

gastrointestinal fluid regulation, pain perception, and inflammation. Many of the current drugs for treatment of Crohn’s disease carry a

greater risk of infection from immunosuppression or allergic reactions, and some must be administered parenterally.

What Is New Here An opioid antagonist, naltrexone 4.5 mg, administered by mouth

once daily significantly improved Crohn’s disease activity index (CDAI) scores and symptoms in subjects with active Crohn’s disease.

Quality of life significantly improved with low-dose naltrexone therapy and remained improved after discontinuation of the drug.

Naltrexone therapy was well tolerated in Crohn’s disease with minimal side effects.

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The use of low doses of naltrexone for the treatment of multiple sclerosis (MS) enjoys a worldwide following amongst MS patients. There is overwhelming anecdotal evidence, that in low doses naltrexone not only prevents relapses in MS but also reduces the progression of the disease. It is proposed that naltrexone acts by reducing apoptosis of oligodendrocytes. It does this by reducing inducible nitric oxide synthase activity. This results in a decrease in the formation of peroxynitrites, which in turn prevent the inhibition of the glutamate transporters. Thus, the excitatory neurotoxicity of glutamate on neuronal cells and oligodendrocytes via activation of the a-amino-3- hydroxy-5-methyl-isoxazole-4-propionic acid class of glutamate receptor is prevented. It is crucial that the medical community respond to patient needs and investigate this drug in a clinical trial.

Agrawal Y.P.Low-dose naltrexone therapy in multiple sclerosis

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Immune Response to Dietary Proteins, Gliadin and Cerebellar Peptides in Children with Autism

Nutritional Neuroscience, 7(3):151-161, 2004Vojdani, T. O’Bryan, J.A. Green, J. McCandless, K.N. Woeller, E. Vojdani, A.A. Nourian, E.L. Cooper

Heat Shock Protein and Gliadin Peptide Promote Development of Peptidase Antibodies in Children with Autism and Patients with Autoimmune Disease

Clin Diag Lab Immunol 11(3):515-524, 2004A. Vojdani, M. Bazargan, E. Vojdani, J. Samadi, A.A. Nourian, N. Eghbalieh, E.L. Cooper

Antibodies to Neuron-Specific Antigens in Childrenwith Autism: Possible Cross-Reaction with Encephalitogenic Proteins from Milk,

Chlamydia pneumoniae and Streptococcus Group AJournal of Neuroimmunology, 129:168-177 2002

A. Vojdani, A.W. Campbell, E. Anyanwu, A. Kashanian, K. Bock, E. Vojdani

Antibodies against Central Nervous System Antigens in Autism: Possible Cross-Reaction with Dietary Proteins and Infectious Agent Antigens.

Neuropsychiatric Disorders & Infections, pp 171-186, 2005, S.H. Fatemi (ed), Taylor & Francis Ltd

Vojdani, T. O’Bryan, J.A. Green, J. McCandless, K.N. Woeller, E. Vojdani, A.A. Nourian, E.L. Cooper

Infections, Toxic Chemicals and Dietary Peptides Binding to Lymphocyte Receptors and Tissue Enzymes are Major Instigators of Autoimmunity in Autism

Int J Immunopathol Pharmacol 16(3):189-199, 2003A. Vojdani, J.B. Pangborn, E. Vojdani, E.L.Cooper

Neuroimmunology

journal of

Low natural killer cell cytotoxic activity in autism: the role of glutathione, IL-2 and IL-15

A. Vojdani, et al.

Although many articles have reported immune abnormalities in autism, NK cell activity has only been examined in one study of 31 patients, of whom 12 were found to have reduced NK activity. The mechanism behind this low NK cell activity was not explored. For this reason, we explored the measurement of NK cell activity in 1027 blood samples from autistic children obtained from ten clinics and compared the results to 113 healthy controls. This counting of NK cells and the measurement of their lytic activity enabled us to express the NK cell activity/100 cells. At the cutoff of 15-50 LU we found that NK cell activity was low in 41-81% of the patients from the different clinics. This NK cell activity below 15 LU was found in only 8% of healthy subjects (p < 0.001). Low NK cell activity in both groups did not correlate with percentage and absolute number of CD16 +/CD56+ cells. When the NK cytotoxic activity was expressed based on activity/100 CD16+/CD56+ cells, several patients who had displayed NK cell activity below 15 LU exhibited normal NK cell activity. Overall, after this correction factor, 45% of the children with autism still exhibited low NK cell activity, correlating with the intracellular level of glutathione. Finally, we cultured lymphocytes of patients with low or high NK cell activity/cell with or without glutathione, IL-2 and IL-15. The induction of NK cell activity by IL-2, IL-15 and glutathione was more pronounced in a subgroup with very low NK cell activity. We conclude that 45% of a subgroup of children with autism suffers from low NK cell activity, and that low intracellular levels of glutathione, IL-2 and IL-15 may be responsible.

“The induction of NK cell activity by IL-2, IL-15 and glutathione was more pronounced in a subgroup with very low NK cell activity. We conclude that 45% of a subgroup of children with autism suffers from low NK cell activity, and that low intracellular levels of glutathione, IL-2 and IL-15 may be responsible.”

Journal of Neuroimmunology (In Press)

Percentage of NK cell activity below 15 lytic units in controls and patients with autism obtained from 10 different clinics.

Vojdani A. et al., Journal of Neuroimmunology 2008 (In Press)

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8%

56%

70%

50%46%

53%

41%

56%

71%64%

81%

0

10

20

30

40

50

60

70

80

90

100

Controls 1 2 3 4 5 6 7 8 9 10

Figure 2 - % NK cell activity below 15 lytic units in controls and patients with autism obtained from 10 different clinics

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Effects of Opioids on the Immune System

Schematic representation of the hematopoietic system showing the differentiation pathways sensitive to opioids.from Effects of Opioids on the Immune System – Roy S. and Loh H.H., Neurochemical Research, 21:1375-1386, 1996

NALTREXONE

High Dose Low Dose

Stimulation of T, B and NK function IFN- and IL-2

production

Inhibition of T, B and NK function IFN- and IL-2

production

-Opioid Receptor Antagonist

-Opioid Receptor Agonist

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MECHANISM OF ACTION OF LDN

Effect on no. of liquid bowel

movements

Healing & repair of mucosal

tissue

Improvement in Crohn’s

disease activity

PromotionOf DNA

synthesis

Healingof

corneal ulcers

Improvement in

inflammatory reaction

Enhancement of

immune function

Down regulation

of TH-17

LDN

Regulation of TReg and

production of IL-10 and

TGF-

Blockade of opiate-R in GI

tract

Interaction of the nuclear opioid growth factor

receptor

Inhibitionof

proinflammatory cytokines

Increase in endogenous enkephalin

and endorphin

The Role of Environmental Factors in Gut-Brain Inflammation and Its Possible Inhibition by LDN

LDN

LDN

LDN

LDNLDN

LDN

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