Potentiation of rat lymphocyte proliferation by novel non-peptidic synthetic opioids

www.elsevier.com/locate/intimp

International Immunopharmaco

Potentiation of rat lymphocyte proliferation by novel non-peptidic

synthetic opioids

Diana Caballero-Hernandeza, Richard J. Weberb, Mary E. Hicksb, Reyes Tamez-Guerraa,

Cristina Rodrıguez-Padillaa, Patricia Tamez-Guerraa, Kenner C. Ricec,

Subramaniam Ananthand, Ricardo Gomez-Floresa,TaDepartamento de Microbiologıa e Inmunologıa, Facultad de Ciencias Biologicas, Universidad Autonoma de Nuevo Leon,

San Nicolas de los Garza, NL, MexicobDepartment of Biomedical and Therapeutic Sciences, UIC College of Medicine, Peoria, IL, United States

cLaboratory of Medicinal Chemistry, NIDDK, NIH, Bethesda, MD, United StatesdOrganic Chemistry Department, Southern Research Institute, Birmingham, Alabama 35255, United States

Received 27 May 2004; received in revised form 23 August 2004; accepted 16 March 2005

Abstract

Opioids represent a major source of relief for acute and chronic, moderate to severe nonmalignant pain. However, opioid abuse

may cause immunosuppression leading to infections and cancer development. Recently we reported results on novel non-peptidic

delta- and mu-selective opioids that induced immunopotentiation in vitro and ex vivo. In the present study, we investigated the

effects of the delta agonist SNC 80, and mu agonists, naltrindole and naltrexone derivatives for their capacity to alter

lymphoproliferation in vitro. They were observed to stimulate lymphoproliferation at concentrations ranging from 10�10 to 10�5

M. SNC 80 significantly ( p b0.05) stimulated (43–311%) proliferation of resident and concanavalin A (Con A)-treated

lymphocytes; the naltrindole derivatives 9332 and 9333 caused significant ( p b0.05) 26–47% and 13–43%, respectively,

stimulation of Con A-treated lymphoproliferation; whereas the naltrexone derivatives 9334 and 9336 significantly ( p b0.05)

stimulated 9–40% and 15–69%, respectively, proliferation of resident and Con A-treated lymphocytes. These novel opioid ligands

could serve as immunotherapeutic agents by increasing the pool of lymphocytes with potential use in the treatment of infectious

diseases including AIDS. This study provides evidence of the relationship structure/function of opioids on lymphoproliferation,

and supports further evaluation of opioids with immunomodulatory potential in preclinical and clinical studies.

D 2005 Elsevier B.V. All rights reserved.

Keywords: SNC 80; Naltrindole derivatives; Naltrexone derivatives; Non-peptide opioids; y-opioid agonist; A-opioid agonists;

Lymphoproliferation

1567-5769/$ - s

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E-mail addr

logy 5 (2005) 1271–1278

ee front matter D 2005 Elsevier B.V. All rights reserved.

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ng author. Loma Panoramica #321-1, Colonia Loma Larga, Monterrey, Nuevo Leon, Mexico, C.P. 64710. Tel.: +52 83 29 41

52 83 52 42 12.

ess: [email protected] (R. Gomez-Flores).

D. Caballero-Hernandez et al. / International Immunopharmacology 5 (2005) 1271–12781272

1. Introduction

There has been recent interest in investigating the

effects of opioids on the immune system, particularly,

if it pertains to the association of infectious diseases

such as AIDS, with intravenous drug abuse. Opioids

include opium, morphine, heroin, codeine, meperi-

dine, and methadone. All of these relax the central

nervous system and have similar sleep-inducing and

narcotic (pain-relieving) effects. Heroin is the most

commonly abused opioid drug in the United States;

there is an estimated 400,000–600,000 heroin addicts

in the U.S. alone. It is known that over time, opioid

users who inject the drug may develop infections of

the heart lining and valves, skin abscesses, and lung

congestion. Infections can lead to hepatitis, tetanus,

liver disease, and HIV transmission [1], and alter-

ations of immune parameters also have been reported

among drug abusers [2–4]. In this respect, immuno-

logic dysfunction in heroin addicts has been docu-

mented since 1974 [5]. In AIDS patients, cell-

mediated immunity is usually impaired; however, in

late stages of the disease, both the cell- and antibody-

mediated immune responses start failing, and lymph

node atrophy results. It is recognized that virtually all

drugs with abuse potential have central actions and

many abused drugs have immunosuppressive effects

leading to infectious diseases [6–11].

A significant reduction in the absolute number and

percentage of total and active T lymphocytes in the

peripheral blood of opiate addicts and T-cell rosette

formation was early reported [12]. Brown et al. (1974)

also reported impaired in vitro lymphoproliferative

responses to the mitogens PHA, concavanalin A, and

pokeweed mitogen in heroin addicts [5], whereas

Brugo et al. (1983) demonstrated a significant

reduction of PHA response of lymphocytes in

methadone patients [13]. In addition, Fecho et al.

(2000) demonstrated that heroin induced a dose-

dependent, naltrexone-reversible suppression of the

concanavalin A-stimulated proliferation of rat T cells

and lipopolysaccharide-stimulated proliferation of B

cells [14]. Furthermore, Wang et al. (2002) showed

that chronic morphine treatment in mice resulted in a

significantly two- to three-fold inhibition of thymic,

splenic, and lymph node cellularity, inhibition of

thymic–lymphocyte proliferation, and inhibition of

IL-2 synthesis [15].

Novel opioid compounds have been synthesized

that have analgesic capacity, but lack immunosup-

pressive effects or even potentiate immune function

[16–19]. In this respect, Nowak et al. (1998) reported

that the non-peptide agonist SNC 80, a y-opioidligand, did not alter NK cell, lymphocyte, and

macrophage functions following intracerebroventricu-

lar administration in Fischer 344N male rats [16].

However, in these animals, intravenous administration

of SNC 80 was associated with ex vivo immunopo-

tentiation, following an activating challenge [18]. In

addition, we reported that novel non-peptide naltrin-

dole derivatives and the tetrahydroquinoline CGPM-9

possess immunoenhancing properties in vitro [17,19].

Additionally, we have observed that heroin self-

administration altered immune function, leading to

evidence of infection, followed by chronic activation

of innate immune function, cachexia, and weight loss

[20].

Although, data may suggest that opioids are

involved in the individual cell-mediated immune

response [5,12,15–20], our understanding of the

effects of opioids on the immune system is incom-

plete. It is necessary to better understand the structure/

function relationship of opioids involved on immu-

nomodulation associated with susceptibility/resistance

to diseases.

The present study was undertaken to evaluate the

in vitro effect of SNC 80, and naltrindole and

naltrexone derivatives on proliferation of rat lympho-

cytes. We observed potent lymphoproliferative activ-

ities of these opioids.

2. Material and methods

2.1. Reagents, culture media, and cell lines

Penicillin–streptomycin solution, l-glutamine,

ficoll-hypaque solution, trypsin–EDTA solution,

and RPMI 1640 media were obtained from Life

Technologies (Grand Island, NY). Fetal bovine

serum (FBS), concanavalin A (Con A), sodium

dodecyl sulfate (SDS), N, N-dimethylformamide

(DMF), PBS, and 3-[4,5-dimethylthiazol-2-yl]-2,5-

diphenyltetrazolium bromide (MTT) were purchased

from Sigma Chemical Co. (St. Louis, MO).

Extraction buffer was prepared by dissolving 20%

D. Caballero-Hernandez et al. / International Immunopharmacology 5 (2005) 1271–1278 1273

(wt /vol) SDS at 37 8C in a solution of 50% each

DMF and demineralized water, and the pH was

adjusted to 4.7.

2.2. Drugs

The synthetic opioids (+)-4-((alpha R) 9-alpha-((2S,

5R)-4-allyl-2, 5-dimethyl-1-piperazinyl)-3-methoxy-

benzil)-N, N-diethyl-benzamide, SNC 80 (+); nal-

trindole derivatives: phenoxynaltrindole (9332) and

6V-Hydroxynaltrindole, (9333); and the benzylidene-

naltrexone derivatives: 6,7-5V, 6V-pyridomorphinan, 17-

(cyclopropylmethyl)-6,7-dehydro-4,5a-epoxy,3,14-di-

hydroxy-1V2V-dihydro-2Voxo-3;-cyano-4Vphenyl (9334)

and 6, 7-2V,3V-quinolinomorphinan, 17-(cyclopropyl-

methyl)-6,7-didehydro-3,14h-dihydro-4,5a-epoxy-4V-phenyl (9336); were provided by K.C. Rice and

Subramaniam Ananthan.

2.3. Animals

Sprague–Dawley male rats (200–220 g) were

purchased from Harlan Sprague–Dawley, Inc. (Indian-

apolis, IN). They were kept in a pathogen- and stress-

free environment at 24 8C, under a light–dark cycle

(light phase, 06:00–18:00 hours), and given water and

food ad libitum. Animals were euthanized by asphyx-

iation in 100% CO2 chamber.

2.4. Cell preparation and culture

Thymus was removed immediately after rat death,

and a single cell-suspension was prepared by disrupt-

ing the organ in RPMI 1640 medium as previously

reported [21]. The cell suspension was washed three

times in this medium, suspended and adjusted to

1�107 cells/ml in AIM-V medium. Because serum

has been reported to potentiate immune function [22],

the culture medium was changed at this step to AIM-

V medium, a serum-free medium capable of support-

ing cell culture [23].

2.5. T cell proliferation assay

T cell proliferation was determined by a colori-

metric technique using MTT [24]. Thymic cell

suspensions (100 Al of 1�107 cells/ml) were added

to flat-bottomed 96-well plates (Becton Dickinson)

containing triplicate cultures (100 Al) of AIM-V

medium (unstimulated control) or opioid derivatives

at various concentrations, in the presence or absence

of 0.6, 1.2 and 2.4 Ag/ml of concanavalin A (the

mitogen was added 6 h prior to the addition of the

opioids, from time–course experiments (data not

shown)) for 48 h at 37 8C in 95% air–5% CO2

atmosphere. After incubation for 44 h at 37 8C with

5% CO2, MTT (0.5 mg/ml, final concentration) was

added, and cultures were additionally incubated for 4

h. Cell cultures were then incubated for 16 h with

extraction buffer (100 Al), and optical densities,

resulting from dissolved formazan crystals, were then

read in a microplate reader (Bio-Tek Instruments, Inc.,

Winooski, VT) at 540 nm. The lymphocyte prolifer-

ation index (LPI) was calculated as follows:

LPI ¼ A540 in opioid-treated cells

A540 in untreated cells

2.6. Statistical analysis

The results were expressed as meanFSEM of the

response of 3 replicate determinations from three

independent experiments. Level of significance was

assessed by Dunnet’s t test.

3. Results

3.1. Effect of non-peptidic opioids on

lymphoproliferation

SNC 80 significantly ( p b0.05) stimulated 73%,

56%, 56%, and 56% proliferation of resident, Con A-

untreated, lymphocytes at 10�8, 10�7, 10�6, and

10�5 M respectively; stimulated 47%, 43%, and 51%

proliferation of Con A (1.2 Ag/ml)-treated lympho-

cytes at 10-9, 10�8, and 10�7 M respectively; and

stimulated 310%, 311%, 290%, and 173% prolifer-

ation of Con A (2.4 Ag/ml)-treated lymphocytes at

10�9, 10�8, 10�7, and 10�6 M respectively, as

compared with SNC 80-untreated control (Fig. 1).

Combination of SNC 80 at 10�5 M plus Con A at 0.6

and 1.2 Ag/ml caused significant 9% and 44% toxicity,

respectively (Fig. 1). The compound 9332 signifi-

cantly ( p b0.05) stimulated 30%, 27%, and 41%

10-11 10-10 10-9 10-8 10-7 10-6 10-50,0

0,5

1,0

1,5

2,0

2,5

3,0

**

** **

****

Lym

phoc

yte

prol

ifera

tion

inde

x UntreatedCon A (0.6 µg/ml)Con A (1.2 µg/ml)Con A (2.4 µg/ml)

9332 (M)

Fig. 2. Lymphoproliferation induced by 9332. Rat thymic cel

suspensions were incubated in the presence or absence of various

concentrations of 9332 and Con A, after which lymphoproliferation

was measured colorimetrically, as explained in the text. Data

represent meansFSE of triplicate determinations from three

independent experiments. *p b0.05 (Dunnet’s test) compared with

9332-untreated control. Lymphocyte proliferation index=A540 in

opioid-treated cells /A540 in untreated cells. Optical density values

for proliferation of 9332-untreated controls were 0.30F0.02

0.41F0.04, 0.42F0.03, and 0.42F0.02 for untreated and 0.6

1.2, and 2.4 Ag/ml Con A-treated cells, respectively.

10-11 10-10 10-9 10-8 10-7 10-6 10-50.0

0.5

1.0

1.5

2.0

**

***

*

Lym

phoc

yte

prol

ifera

tion

inde

x Untreated controlCon A 0.6 µg/mlCon A 1.2 µg/mlCon A 2.4 µg/ml

9333 (M)












10-11 10-10 10-9 10-8 10-7 10-6 10-50

1

2

3

4

5

6

*

**

**** ***

***

Lym

phoc

yte

prol

ifera

tion

inde

x UntreatedCon A (0.6 µg/ml)Con A (1.2 µg/ml)Con A (2.4 µg/ml)

SNC 80 (M)

Fig. 1. Lymphoproliferation induced by SNC 80. Rat thymic cell


concentrations of SNC 80 and Con A, after which lymphoprolifera-

tion was measured colorimetrically, as explained in the text. Data



SNC 80-untreated control. Lymphocyte proliferation index=A540 in


for proliferation of SNC 80-untreated controls were 0.17F0.05,

0.28F0.09, 0.53F0.20, and 0.35F0.12 for untreated and 0.6, 1.2,

and 2.4 Ag/ml Con A-treated cells, respectively.


proliferation of Con A (1.2 Ag/ml)-treated lympho-

cytes at 10�8, 10�7, and 10 �6 M, respectively; and

stimulated 26%, 34%, 41%, 46%, and 47% prolifer-

ation of Con A (2.4 Ag/ml)-treated lymphocytes at

10�10, 10�9, 10�8, 10�7, and 10�6 M respectively,

as compared with 9332-untreated control (Fig. 2). The

opioid 9332 at 10�5 M alone or in combination with

Con A at 0.6, 1.2, and 2.4 Ag/ml caused 55%, 59%,

58%, and 55% toxicity, respectively (Fig. 2). The

compound 9333 significantly ( p b0.05) stimulated

13%, 24%, 35%, 37%, 43%, and 28% proliferation of

Con A (2.4 Ag/ml)-treated lymphocytes at 10�10,

10�9, 10�8, 10�7, 10�6, and 10�5 M, respectively, as

compared with 9333-untreated control (Fig. 3). The

opioid 9334 significantly ( pb0.05) stimulated 16%,

21%, 17%, 22%, 19%, and 14% proliferation of

resident, Con A-untreated lymphocytes; stimulated


Con A (0.6 Ag/ml)-treated lymphocytes; stimulated


Con A (1.2 Ag/ml)-treated; and stimulated 22%, 36%,

39%, 40%, 35%, and 34% proliferation of Con A (2.4

Ag/ml)-treated lymphocytes, at 10�10, 10�9, 10�8,

10�7, 10�6, and 10�5 M, respectively, as compared

with 9334-untreated control (Fig. 4); and 9336

l

,

,

significantly ( p b0.05) stimulated 15%, 25%, 24%,

24%, and 25% proliferation of resident, Con A-

untreated lymphocytes at 10�10, 10�9, 10�8, 10�7,

l

,

,

10-11 10-10 10-9 10-8 10-7 10-6 10-50.0

0.5

1.0

1.5

2.0

2.5

**** **** **

**** **** **

**** ***

* ***

Lym

phoc

yte

prol

ifera

tion

inde


9336 (M)













and 10�6 M respectively; stimulated 19%, 19%, 24%,

28%, 45%, and 62% proliferation of Con A (0.6 Ag/ml)-treated lymphocytes at 10�10, 10�9, 10�8, 10�7,

10�6, and 10�5 M respectively; stimulated 25%,

31%, 32%, 51%, and 62% proliferation of Con A (1.2

Ag/ml)-treated at 10�9, 10�8, 10�7, 10�6, and 10�5

M, respectively; and stimulated 21%, 48%, 51%,

48%, 69%, and 54% proliferation of Con A (2.4 Ag/ml)-treated lymphocytes, at 10�10, 10�9, 10�8, 10�7,

10�6, and 10�5 M, respectively, as compared with

9336-untreated control (Fig. 5).

3.2. Effect of Con A on opioid-induced

lymphoproliferation

The use of Con A at 2.4 Ag/ml was observed to

increase the lymphoproliferative response to opioids.

In this respect, Con A significantly ( p b0.05) increased

SNC 80-induced lymphoproliferation at concentrations

ranging from 10�11 to 10�6 M, as compared with SNC

80-treated control response (Fig. 1), whereas Con A at

1.2 Ag/ml, significantly ( p b0.05) increased 9332-

induced lymphoproliferation at concentrations of

10�8, 10�7, and 10�6 M, as compared with 9332-

treated control response (Fig. 2). Similarly, Con A at

10-11 10-10 10-9 10-8 10-7 10-6 10-50.0

0.5

1.0

1.5

2.0

2.5

**********************

*****

***

****

Lym

phoc

yte

prol

ifera

tion

inde


9334 (M)

Fig. 4. Lymphoproliferation induced by 9334. Rat thymic cell








for proliferation of 9334-untreated controls were 0.26F0.01,

0.43F0.04, 0.49F0.03, and 0.52F0.02 for untreated and 0.6,


l

,

,

2.4 Ag/ml, significantly ( pb0.05) increased 9332-

induced lymphoproliferation at concentrations ranging

from 10�10to 10�6, as compared with 9332-treated

control response (Fig. 2). In addition, Con A at 2.4 Ag/ml, significantly ( p b0.05) increased 9333-induced

lymphoproliferation at concentrations of 10�10–10�6

M, as compared with 9333-treated control response

(Fig. 3). Furthermore, Con A at 2.4 Ag/ml, significantly

( p b0.05) increased 9334-induced lymphoprolifera-

tion at 10�8, as compared with 9334-treated control

response (Fig. 4). In addition, Con A at 0.6 Ag/ml,

significantly ( p b0.05) increased 9336-induced lym-

phoproliferation at concentrations of 10�6 and 10�5

M, as compared with 9336-treated control response

(Fig. 5). Similarly, Con A at 2.4 Ag/ml, significantly

( p b0.05) increased 9336-induced lymphoprolifera-

tion at concentrations ranging from 10�9 to 10�5,

as compared with 9336-treated control response

(Fig. 5).

4. Discussion

In the present study, we demonstrated the potential

of mu and delta non-peptidic opioids to stimulate

lymphocyte proliferation. We observed that SNC 80


was more potent than the naltrindole and naltrexone

derivatives to stimulate lymphoproliferation (up to

311%, 47%, and 69% increases, respectively,

compared with untreated control). Opioid-mediated

lymphoproliferation was partially reversed by the

opioid antagonists CTOP and naltrexone (data not

shown). The concept of functionally coupled A and

y opioid receptors might explain the immunoenhanc-

ing effect of these opioids as well as the ability of

CTOP and naltrexone to partially block their effects.

Since these compounds are selective but not specific

for a type of opioid receptor, more investigation is

needed to determine their mechanism of action. In

addition, we showed that Con A significantly

potentiated opioid-induced lymphocyte proliferation

(Figs. 1–5). Stimulation of proliferation of rat

thymocytes in vitro has been reported to be

triggered by suboptimal concentrations of Con A

[25]. Potentiation of Con A-induced proliferation by

opioids may be due to upregulation of opioid

receptors on these cells. Although this remains to

be elucidated, others have shown that mu-, delta-

and kappa-opioid receptor selective agonists are

potent in vitro stimulators of mitogen-induced

proliferation of murine T lymphocytes [26], which

has been suggested to be related to upregulation of

opioid receptor expression upon lymphocyte activa-

tion by Con A [27,28]. We have previously reported

that intravenous administration of SNC 80 was

associated with ex vivo immunopotentiation, follow-

ing an activating challenge with Con A [18];

however, the ex vivo synergistic effect of naltrindole

and naltrexone derivatives plus a lymphocyte-acti-

vating agent remains to be elucidated.

Opioids are recognized for modulating some

aspects of immune function [6–10]. A number of

research groups have importantly contributed to the

growing pool of information suggesting direct and

indirect roles of opioids and immune function,

particularly as this association relates to infectious

diseases [29–31]. On this regard, morphine and

methadone have been reported to enhance HIV

infection of macrophages through the downregula-

tion of beta-chemokine production and upregulation

of CCR5 receptor expression [29,30]. The immu-

nosuppressive effects of the morphine following in

vivo administration were early shown to be

mediated by opioid receptors found within the

central nervous system [6,8,11,32]. Although the

major effect of strong mu agonists in vivo is

immunosuppressive and indirect, direct effects in

vitro of peptidic and novel non-peptidic opioids are

well substantiated [21,32]. Paradoxically, the direct

effect of certain opioids on leukocytes can enhance,

suppress, or have no effect on in vitro and in vivo

parameters of immune function. Eisenstein et al.,

have previously demonstrated that opioids directly

affect cellular and humoral immune functions

though classical opioid receptors [33]. This research

group has shown that mu, kappa, and delta opioid

receptors were associated with regulating of lym-

phoid cell production of antibodies [34]. In

addition, it was shown that mu-, delta-, and

kappa-opioid agonists can stimulate chemotaxis in

T lymphocytes which is crucial in inflammatory

processes [9,35]. Novel opioid compounds have

been synthesized that have analgesic capacity, but

lack immunosuppressive effects or even potentiate

immune function [16–19]. Our previous have

shown that tyrosylamido-6-benzyl-1,2,3,4 tetrahy-

droquinoline (CGPM-9) enhanced rat thymic lym-

phocyte proliferative response to concanavalin A, in

a CTOP-reversible manner [19]. More recently, we

observed that morphinans with substituted pyrimi-

dine (methyl, phenyl, hydroxyl, and amino groups)

and pyrazole groups potentiated Con A-induced

thymic cell proliferation (unpublished observations).

Additionally, Nowak et al., (1998) reported that the

delta opioid agonist SNC 80 did not alter NK cell,

lymphocyte, and macrophage functions following

intracerebroventricular administration [16]. How-

ever, intravenous administration of SNC 80 was

associated with ex vivo immunopotentiation, fol-

lowing an activating challenge [18]. Sharp et al.

demonstrated that SNC 80 inhibited HIV-1 expres-

sion, probably acting on delta opioid receptors on T

cells [36]. The same research team recently showed

the adjuvant properties of naltrexone in potentiating

retroviral drugs-induced anti-HIV activity in CD4+

lymphocyte cultures [37]. These in vitro findings

supported the therapeutic potential of opioids for

treating patients with acquired immunodeficiency

syndrome.

In contrast to the observed immunopotentiating

effect of SNC 80, naltrindole and naltrexone

derivatives in the present study, D’Ambrosio et al.


(2004), recently showed that naltrindole derivatives

at 10�5 M, suppressed proliferation of human

lymphocytes stimulated with mitogens, the antigen

PPD, the anti-CD3 monoclonal antibodies, the

superantigen Staphylococcus aureus Cowan strain

1, and alloantigens in the mixed lymphocyte [38].

Our experimental differences may be mainly related

to the type of opioids, the source of lymphocytes,

the type and concentration of the opioid and co-

stimulatory agents, and the cell stimulation period.

Interestingly, we found that combination of SNC 80

at 10�5 M plus Con A (Fig. 1), and the opioid

9332 at 10�5 M alone or in combination with Con

A (Fig. 2), were significantly toxic for lympho-

cytes. Clinical consequences of opioid-mediated

immunosuppression directly affects incidence of

infections, particularly in intravenous opioid addicts

[5].

The diversity of the opioid pharmacopeia,

expanding knowledge of opioid receptor structure

and ligand binding properties, and current expertise

in opioid medicinal chemistry, have provided us

with compounds with immunotherapeutic potential,

as well as pharmacologic tools for investigation of

how the immune system is naturally controlled and

regulated through opiatergic mechanisms. The clin-

ical use of properly designed and synthesized

opioid ligands could serve as immunotherapeutic

agents with potential use in the treatment of

infectious diseases including AIDS, and cancer. In

addition, because surgical stress also induces

immune dysfunction, the search for analgesic drugs

devoid of immunosuppressive effects is of import.

It is clear that knowledge of how opioids produce

direct effects on the immune system may allow the

discovery, design and synthesis of new opioids that

have specific immunoregulatory properties. Future

research should then provide a clearer understand-

ing of the cellular and molecular targets of opioid

action within the immune system, as well as

intracellular signaling activity [39]. The develop-

ment of highly selective, site-specific designer

drugs and innovative gene-therapies may enhance

opioid function, and suppress negative effects on

immune function and drug dependence. Therapeutic

intervention targeted on the opioid pathways will

potentially enrich the quality of life of suffering

individuals.

Acknowledgment

This study was supported by grants I-32914-N and

CN-285-00 (PAICYT) from Consejo Nacional de

Ciencia y Tecnologıa, and Universidad Autonoma

de Nuevo Leon, Mexico, respectively.

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Potentiation of rat lymphocyte proliferation by novel non-peptidic synthetic opioids

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