- Email: [email protected]
Effects of cannabinoid receptor ligands on LPS-induced pulmonary inflammation in mice
Life Scicnq Vol. 63,No. 8, pp. PL 12.5-129,1996 cbpytightQ 1998ekevicr sc*aec Inc. Printedin the WA. All rightsmssrved 0024-32m/98 $19.00t .oo
PI1 SOO24-3205(98)00324-S
ELSEVIER
PH4Riu4coLoGY LEYtTERs Accelerated Communication EFFECTS OF CANNABINOID PULMONARY E. Berdyshev*,
RECEPTOR LIGANDS ON LPS-INDUCED INFLAMMATION IN MICE
E. Boichot, M. Corbel, N. Germain and V Lagente
INSERM U456, Laboratoire de Pharmacodynamie et de Pharmacologic Moleculaire, Faculte des Sciences Pharmaceutiques et Biologiques, Universite de Rennes 1, Rennes, France and (*) Institute of Marine Biology, 690041 Vladivostok, Russia. (Submitted March 31,199& accepted April 20,1998; received in fmal form June 4,1998)
Abstract : The effects of cannabinoid receptor agonists WIN 55,212-2, A9(anandamide) and (A9-THC), arachidonoylethanolamide tetrahydrocannabinol on lipopolysaccharide (LPS) -induced bronchopulmonary palmitoylethanolamide inflammation in mice were investigated. WIN 55,212-2 and A9-THC induced a concentration-dependent decrease in TNF-ar. level in the bronchoalveolar lavage fluid (BALF) (maximum inhibition 52.7% and 36.9% for intranasal doses of 750 nmol.kg-1 and 2.65 mmol.kg-1, respectively). This effect was accompanied by moderately reduced neutrophil recruitment. Palmitoylethanolamide (750 nmol.kg-l) diminished the level of TNF-a in BALF by 31.5% but had no effect on neutrophil recruitment. Anandamide (7.5750 nmol.kg-1) did not influence the inflammatory process but TNF-a level and neutrophil recruitment were decreased by 28.0% and 62.0%, respectively, with 0.075 nmol.kg-1. These results demonstrate that the cannabinoid receptor ligands inhibited LPSinduced pulmonary inflammation and suggest that this effect could be at least in part mediated bv the cannabinoid CBz receptor.0 1998 Elsevier ScienceInc. Key Words: camxhmoid receptor, inflammation, TNP-a, neutrophil, lung, lipolysaccharide, mouse
Introduction Cannabinoids are known to modulate immune cell activity both in vitro and in viva. The action of cannabinoids is mainly immunosuppressive and is described as being mediated by the peripheral cannabinoid CBz receptor. In vitro, the action of cannabinoids is characterized by inhibition of cytotoxic T-cell activity, macrophage activation, lymphocyte proliferation and alteration of the cytokine network (for review see l-3). In viva, cannabinoids reduce general resistance to bacterial or viral infections (4). In spite of the increasing interest in the immunomodulatory properties of cannabinoids, little is known about the modulation of inflammatory response by cannabinoids in viuo. In this study, we investigated the effects of various cannabinoid receptor agonists on the pulmonary inflammatory response induced by inhalation of endotoxin (LPS) in mice. Corresponding author : Prof. Vincent Lagente, INSERM U456, Laboratoire de Pharmacodynamie et de Pharmacologic Moleculaire, Faculte des Sciences Pharmaceutiques et Biologiques, Universite de Rennes 1,2 avenue du Pr. Leon Bernard, 35043 Rennes Cedex, France, tel: 33 (0) 2 99 33 68 50, fax: 33 (0) 2 99 33 62 42.
PL126
CannaKmoid Ligaods and Pulmonary Inflammation
Vol. 63, No. 8, 1998
Methods Male Balb/c mice aged 8 weeks were slightly anaesthetized with ether. The appropriate concentrations of compounds or vehicle were administered intranasally in a total volume of 50 ul. Thirty min later, the animals were exposed for 1 h to an aerosol of 100 ugml-i of endotoxin (lipolysaccharide of E. coli) (LPS, Sigma, St Louis, MO, USA) in saline (NaCI, 0.9%). A group of mice were only exposed to a saline solution (negative control group). Twenty-four h after LPS aerosol exposure, the animals were anaesthetized using sodium pentobarbitone (60 mg.kg-I), their tracheae were cannulated and their lungs washed 10 times with 0.5 ml saline-EDTA (0.9% NaCl in 2.6 M EDTA). The first ml of the bronchoalveolar lavage fluid (BALF) was collected separately, centrifuged at 600 g for 8 min at 4”C, and the supematant was collected for TNF-a determination by specific ELISA (Genzyme diagnostics, Cambridge, MA, USA). Precipitated cells were combined with the second part (4 ml) of the BALF and centrifuged at 600 g for 8 min at 4°C. After lysis of the erythrocytes, the cells were resuspended in 0.5 ml RPMI-1640 supplemented with 1% penicillin, 1% streptomycin, and 1% L-glutamine and the number of total cells was determined using a haemacytometer. BAL cell composition was determined microscopically after cytocentrifugation (Microm, France) May Gri.inwald-Giemsa. Delta-9(600 g for 10 min) and stained using tethahydrocannabinol (A9-THC, Sigma), anandamide (Sigma), WIN 55,212-2 (RBI, Natick, MA, USA) and Palmitoylethanolamide (Institute of Marine Biology, Vladivostok, Russia) were dissolved in ethanol and then diluted in saline just before intranasal administration. Control mice were treated by the vehicle only. Statistical analysis was performed by the ANOVA test and Student’s t-test for independent samples. Significance was assigned when P < 0.05. Results Compared with negative control mice, LPS aerosol induced a marked recruitment of neutrophils in BALF. WIN 55,212-2 (7.5 nmol.kg-1) significantly reduced neutrophil recruitment by 59% but 75 nmol.kg-1 and 0.075 nmolkg-i WIN 55,212-2 did not elicit any significant effect (table 1). A9-THC also inhibited neutrophil recruitment but only at a dose of 2650 nmol.kg-1. Anandamide elicited a 62% inhibitory effect at the lowest dose of 0.075 nmol.kg1 but did not have any effect at higher concentrations. No significant alterations of the number of cells was observed under the treatment of the mice with palmitoylethanolamide LPS aerosol also induced an important enhancement in TNF-cr level in BALF (438.8t-37.4 in comparison to 60.8 ti.7 in negative control mice). Cannabinoid agonists and fatty acid ethanolamides diminished the level of TNF-a in BALF (Figure 1). WIN 55,212-2 displayed the most pronounced and concentration-dependent inhibition of LPS induced TNF-a production (by 52.7%, 42.6% and 33.8% with 750 nmol.kg*,7.5 nmol.kg-* and 0.075 nmol.kg-l, respectively). A9-THC also induced a dose-dependent decrease in TNF-a in BALF with maximum inhibition at the dose of 2650 nmolkg-*. At 750 and 7.5 nmol.kg-i anandamide had no effect on TNF-a in BALF, but at the lowest dose of 0.075 nmol.kg-i reduced it by 28% (P < 0.05). Palmitoylethanolamide also induced a decrease in TNF-a in BALF at 750 nmol.kg1.
PC127
Cannabmoid L&A& and Pulmomuy Inflammation
Vol. 63, No. 8,1998
TABLE 1 Influence of WIN 55212-2, A9-TI-IC, anandamide and palmitoylethanolamide on LPS-induced changes in cell composition in BALF (cells x10 f SEM). * P < 0.05. n : number of mice. Dose
Drugs
n
Total cells
hmol.kg*)
Mononuclear cells
Neutrophils
Negative control
12
393k46
387k40
2kO.8
Positive control
18
1208+_137
128+30
1075+_129
WIN 55,212-2
750 7.5 0.075
4 4 5
835_+155 578f71* 1160-+172
184*88 130+49 306+85
648_+116 445+50* 842112
A9-THC
2650 7.50 130 7.5 0.075
3 4 3 5 5
485k20* 788k80 677f86 1346k242 1256zk382
78.5+15 151+34 150f35 212_+75 llOf12
400+37* 637k53 522151 1134+181 1119+350
Anandamide
750 7.5 0.075
5 5 5
1205X2-223 1412f189 507k118
119+33 108k22 97ti4
1071+193 1292+182 409k94
Palmitoylethanolamide
750 7.5
5 5
993k3.317 1096k252
73+16 106+_62
915k306 988+_192
500 c
T
none
0075
7.5
130
750
LMIU-STHC
2650
7.5 Pamn
750
0.075
75
anandsmide
750
0.075
75
WIN 55212
FIG. 1 and anandamide A9-TI-IC, 55,212-2, Influence of WIN palmitoylethanolamide on the level of TNF-a in the bronchoalveolar lavage fluid (pg,ml-I-+ SEM). * P < 0.05, ** P < 0.01 (n=3-18).
750
PL128
CannabiioidL.igandsand PulmonaryInflammation
Vol. 63, No. 8, 1998
Discussion Inhalation of LPS induced a clear inflammatory response in the lung of mice, which was characterized by massive neutrophil recruitment and release of TNF-o in BALF. Our results demonstrate that WIN 55,212-2 and A9-THC, significantly reduced LPS-induced neutrophil recruitment as well as the level of TNF-a. WIN 55212-2 seems to be a more powerful compound than A9-THC, at least in inhibiting TNF-a synthesis. This is in agreement with binding experiments which have demonstrated that the affinity of WIN 55,212-2, to the cannabinoid CBz receptor is lo-times higher than that of A9-THC (5) and supports the fact that mouse macrophages express predominantly the cannabinoid CBz receptor as human immune tissues and cells do (6). Previously, cannabinoids have been shown to inhibit in vitro the synthesis of TNFa by the mouse macrophage cell line RAW264.7 (7) and the production of TNFa, interleukin-6 and interleukin-8 by human peripheral blood mononuclear cells (8). At present, it is difficult to determine whether or not the observed effect of WIN 55,212-2 and A9-THC results from a specific activation of the cannabinoid CBz receptor, and which part of this effect is determined by a possible activation of the cannabinoid CBi receptor expressed in small quantities by immune cells (6). Hence, a recent report suggest that WIN 55,212-2 may also bind to a non-cannabinoid binding site (9). The development of specific CBz receptor antagonists or binding antibodies should clarify the question of CBz/CBi receptor- mediated modulation of immune response by cannabinoids. and anandamide also demonstrated a weak antiPalmitoylethanolamide inflammatory activity. However, their action was not as clear as that of cannabinoids : only the lowest dose of anandamide reduced the level of TNF-a in BALF and neutrophil recruitment, whereas, palmitoylethanolamide decreased the level of TNF-cx only at the highest micromolar dose but failed to influence neutrophil recruitment. We can suppose that a possible distinct affinity of the compounds for mouse macrophage cannabinoid CBp/CBi receptors, together with their differential hydrolysis by amidase, may be responsible for the observed discrepancy of the action of these two fatty acid ethanolamides. We cannot either dismiss a differential triggering by anandamide and palmitoylethanolamide of subsequent signaling mechanisms as was shown for rat basophilic leukaemia RBL-2H3 cells (10). Thus, the concrete background of anandamide and palmitoylethanolamide interference with bronchopulmonary inflammatory response in mice needs further clarification. In conclusion, we have demonstrated that different agonists of cannabinoid receptors can downmodulate bronchopulmonary inflammation in mice and suggest that this effect of cannabinoid receptor agonists could be mediated at least in part by cannabinoid CBz receptor. Acknowledgement manuscript.
1. 2. 3.
: The authors
than G. Bouer for his assistance
in preparing
the
References BERDYCHEV, EV., E. BOICHOT and V. LAGENTE J. Lipid Mediators Cell Signalling. 15 49-67 (1996). KAMINSKI NE. Biochem. Pharmacol. 52 1133-l 140 (1996). KLEIN TW., C. NEWTON, W. ZHU, Y. DAAKA and H. FRIEDMAN. Proc. Sot. Exp. Biol. Med. 209 205-212 (1995).
Vol. 63,No. 8,1!998
4. 5. 6.
7. 8. 9. 10.
CamubmoidLigandsand PulmonaryInflammation
PL129
MORAHAN, PS., PC. KLYKKEN, SH. SMITH, LS. HARRIS and AE. MUNSON. Infect. Immun. 23 670-674 (1979). FELDER CC., KE. JOYCE, EM. BRILEY, J. MANSOURI, K. MACKIE, 0. BLOND, Y. LAI, AL. MA and RL. MITCHELL. Mol. Pharmacol. 48 443-450 (1995). GALIEGUE S., S. MARY, J. MARCHAND, D. DUSSOSSOY, D. CARRIERE, P. CARAYON, M. BOUABALA, D. Shire, G. LEFUR and P. CASSELLAS. Eur. J. Biochem. 232 54-61 (1995). FISCHER-STENGER K., DAD. PETTIT and GA. CABRAL. J. Pharmacol. Exp. Ther. 267 1558-1565 (1993). BERDYCHEV EV., E. BOICHOT, N. GERMAIN, N. ALLAIN, JP. ANGER and V. LAGENTE Eur. J. Pharmacol. 330 231-240 (1997). STARK, S., MA. PACHECO and SR. CHILDERS. Cell. Mol. Neurobiol. 17 483-493 (1997). FACCI L., R. DAL TOSO, S. ROMANELLO, A. BURIANI, SD. SKAPER and A. LEON. Proc. Natl. Acad. Sci. USA 92 3376-3380 (1995).
PI1 SOO24-3205(98)00324-S
ELSEVIER
PH4Riu4coLoGY LEYtTERs Accelerated Communication EFFECTS OF CANNABINOID PULMONARY E. Berdyshev*,
RECEPTOR LIGANDS ON LPS-INDUCED INFLAMMATION IN MICE
E. Boichot, M. Corbel, N. Germain and V Lagente
INSERM U456, Laboratoire de Pharmacodynamie et de Pharmacologic Moleculaire, Faculte des Sciences Pharmaceutiques et Biologiques, Universite de Rennes 1, Rennes, France and (*) Institute of Marine Biology, 690041 Vladivostok, Russia. (Submitted March 31,199& accepted April 20,1998; received in fmal form June 4,1998)
Abstract : The effects of cannabinoid receptor agonists WIN 55,212-2, A9(anandamide) and (A9-THC), arachidonoylethanolamide tetrahydrocannabinol on lipopolysaccharide (LPS) -induced bronchopulmonary palmitoylethanolamide inflammation in mice were investigated. WIN 55,212-2 and A9-THC induced a concentration-dependent decrease in TNF-ar. level in the bronchoalveolar lavage fluid (BALF) (maximum inhibition 52.7% and 36.9% for intranasal doses of 750 nmol.kg-1 and 2.65 mmol.kg-1, respectively). This effect was accompanied by moderately reduced neutrophil recruitment. Palmitoylethanolamide (750 nmol.kg-l) diminished the level of TNF-a in BALF by 31.5% but had no effect on neutrophil recruitment. Anandamide (7.5750 nmol.kg-1) did not influence the inflammatory process but TNF-a level and neutrophil recruitment were decreased by 28.0% and 62.0%, respectively, with 0.075 nmol.kg-1. These results demonstrate that the cannabinoid receptor ligands inhibited LPSinduced pulmonary inflammation and suggest that this effect could be at least in part mediated bv the cannabinoid CBz receptor.0 1998 Elsevier ScienceInc. Key Words: camxhmoid receptor, inflammation, TNP-a, neutrophil, lung, lipolysaccharide, mouse
Introduction Cannabinoids are known to modulate immune cell activity both in vitro and in viva. The action of cannabinoids is mainly immunosuppressive and is described as being mediated by the peripheral cannabinoid CBz receptor. In vitro, the action of cannabinoids is characterized by inhibition of cytotoxic T-cell activity, macrophage activation, lymphocyte proliferation and alteration of the cytokine network (for review see l-3). In viva, cannabinoids reduce general resistance to bacterial or viral infections (4). In spite of the increasing interest in the immunomodulatory properties of cannabinoids, little is known about the modulation of inflammatory response by cannabinoids in viuo. In this study, we investigated the effects of various cannabinoid receptor agonists on the pulmonary inflammatory response induced by inhalation of endotoxin (LPS) in mice. Corresponding author : Prof. Vincent Lagente, INSERM U456, Laboratoire de Pharmacodynamie et de Pharmacologic Moleculaire, Faculte des Sciences Pharmaceutiques et Biologiques, Universite de Rennes 1,2 avenue du Pr. Leon Bernard, 35043 Rennes Cedex, France, tel: 33 (0) 2 99 33 68 50, fax: 33 (0) 2 99 33 62 42.
PL126
CannaKmoid Ligaods and Pulmonary Inflammation
Vol. 63, No. 8, 1998
Methods Male Balb/c mice aged 8 weeks were slightly anaesthetized with ether. The appropriate concentrations of compounds or vehicle were administered intranasally in a total volume of 50 ul. Thirty min later, the animals were exposed for 1 h to an aerosol of 100 ugml-i of endotoxin (lipolysaccharide of E. coli) (LPS, Sigma, St Louis, MO, USA) in saline (NaCI, 0.9%). A group of mice were only exposed to a saline solution (negative control group). Twenty-four h after LPS aerosol exposure, the animals were anaesthetized using sodium pentobarbitone (60 mg.kg-I), their tracheae were cannulated and their lungs washed 10 times with 0.5 ml saline-EDTA (0.9% NaCl in 2.6 M EDTA). The first ml of the bronchoalveolar lavage fluid (BALF) was collected separately, centrifuged at 600 g for 8 min at 4”C, and the supematant was collected for TNF-a determination by specific ELISA (Genzyme diagnostics, Cambridge, MA, USA). Precipitated cells were combined with the second part (4 ml) of the BALF and centrifuged at 600 g for 8 min at 4°C. After lysis of the erythrocytes, the cells were resuspended in 0.5 ml RPMI-1640 supplemented with 1% penicillin, 1% streptomycin, and 1% L-glutamine and the number of total cells was determined using a haemacytometer. BAL cell composition was determined microscopically after cytocentrifugation (Microm, France) May Gri.inwald-Giemsa. Delta-9(600 g for 10 min) and stained using tethahydrocannabinol (A9-THC, Sigma), anandamide (Sigma), WIN 55,212-2 (RBI, Natick, MA, USA) and Palmitoylethanolamide (Institute of Marine Biology, Vladivostok, Russia) were dissolved in ethanol and then diluted in saline just before intranasal administration. Control mice were treated by the vehicle only. Statistical analysis was performed by the ANOVA test and Student’s t-test for independent samples. Significance was assigned when P < 0.05. Results Compared with negative control mice, LPS aerosol induced a marked recruitment of neutrophils in BALF. WIN 55,212-2 (7.5 nmol.kg-1) significantly reduced neutrophil recruitment by 59% but 75 nmol.kg-1 and 0.075 nmolkg-i WIN 55,212-2 did not elicit any significant effect (table 1). A9-THC also inhibited neutrophil recruitment but only at a dose of 2650 nmol.kg-1. Anandamide elicited a 62% inhibitory effect at the lowest dose of 0.075 nmol.kg1 but did not have any effect at higher concentrations. No significant alterations of the number of cells was observed under the treatment of the mice with palmitoylethanolamide LPS aerosol also induced an important enhancement in TNF-cr level in BALF (438.8t-37.4 in comparison to 60.8 ti.7 in negative control mice). Cannabinoid agonists and fatty acid ethanolamides diminished the level of TNF-a in BALF (Figure 1). WIN 55,212-2 displayed the most pronounced and concentration-dependent inhibition of LPS induced TNF-a production (by 52.7%, 42.6% and 33.8% with 750 nmol.kg*,7.5 nmol.kg-* and 0.075 nmol.kg-l, respectively). A9-THC also induced a dose-dependent decrease in TNF-a in BALF with maximum inhibition at the dose of 2650 nmolkg-*. At 750 and 7.5 nmol.kg-i anandamide had no effect on TNF-a in BALF, but at the lowest dose of 0.075 nmol.kg-i reduced it by 28% (P < 0.05). Palmitoylethanolamide also induced a decrease in TNF-a in BALF at 750 nmol.kg1.
PC127
Cannabmoid L&A& and Pulmomuy Inflammation
Vol. 63, No. 8,1998
TABLE 1 Influence of WIN 55212-2, A9-TI-IC, anandamide and palmitoylethanolamide on LPS-induced changes in cell composition in BALF (cells x10 f SEM). * P < 0.05. n : number of mice. Dose
Drugs
n
Total cells
hmol.kg*)
Mononuclear cells
Neutrophils
Negative control
12
393k46
387k40
2kO.8
Positive control
18
1208+_137
128+30
1075+_129
WIN 55,212-2
750 7.5 0.075
4 4 5
835_+155 578f71* 1160-+172
184*88 130+49 306+85
648_+116 445+50* 842112
A9-THC
2650 7.50 130 7.5 0.075
3 4 3 5 5
485k20* 788k80 677f86 1346k242 1256zk382
78.5+15 151+34 150f35 212_+75 llOf12
400+37* 637k53 522151 1134+181 1119+350
Anandamide
750 7.5 0.075
5 5 5
1205X2-223 1412f189 507k118
119+33 108k22 97ti4
1071+193 1292+182 409k94
Palmitoylethanolamide
750 7.5
5 5
993k3.317 1096k252
73+16 106+_62
915k306 988+_192
500 c
T
none
0075
7.5
130
750
LMIU-STHC
2650
7.5 Pamn
750
0.075
75
anandsmide
750
0.075
75
WIN 55212
FIG. 1 and anandamide A9-TI-IC, 55,212-2, Influence of WIN palmitoylethanolamide on the level of TNF-a in the bronchoalveolar lavage fluid (pg,ml-I-+ SEM). * P < 0.05, ** P < 0.01 (n=3-18).
750
PL128
CannabiioidL.igandsand PulmonaryInflammation
Vol. 63, No. 8, 1998
Discussion Inhalation of LPS induced a clear inflammatory response in the lung of mice, which was characterized by massive neutrophil recruitment and release of TNF-o in BALF. Our results demonstrate that WIN 55,212-2 and A9-THC, significantly reduced LPS-induced neutrophil recruitment as well as the level of TNF-a. WIN 55212-2 seems to be a more powerful compound than A9-THC, at least in inhibiting TNF-a synthesis. This is in agreement with binding experiments which have demonstrated that the affinity of WIN 55,212-2, to the cannabinoid CBz receptor is lo-times higher than that of A9-THC (5) and supports the fact that mouse macrophages express predominantly the cannabinoid CBz receptor as human immune tissues and cells do (6). Previously, cannabinoids have been shown to inhibit in vitro the synthesis of TNFa by the mouse macrophage cell line RAW264.7 (7) and the production of TNFa, interleukin-6 and interleukin-8 by human peripheral blood mononuclear cells (8). At present, it is difficult to determine whether or not the observed effect of WIN 55,212-2 and A9-THC results from a specific activation of the cannabinoid CBz receptor, and which part of this effect is determined by a possible activation of the cannabinoid CBi receptor expressed in small quantities by immune cells (6). Hence, a recent report suggest that WIN 55,212-2 may also bind to a non-cannabinoid binding site (9). The development of specific CBz receptor antagonists or binding antibodies should clarify the question of CBz/CBi receptor- mediated modulation of immune response by cannabinoids. and anandamide also demonstrated a weak antiPalmitoylethanolamide inflammatory activity. However, their action was not as clear as that of cannabinoids : only the lowest dose of anandamide reduced the level of TNF-a in BALF and neutrophil recruitment, whereas, palmitoylethanolamide decreased the level of TNF-cx only at the highest micromolar dose but failed to influence neutrophil recruitment. We can suppose that a possible distinct affinity of the compounds for mouse macrophage cannabinoid CBp/CBi receptors, together with their differential hydrolysis by amidase, may be responsible for the observed discrepancy of the action of these two fatty acid ethanolamides. We cannot either dismiss a differential triggering by anandamide and palmitoylethanolamide of subsequent signaling mechanisms as was shown for rat basophilic leukaemia RBL-2H3 cells (10). Thus, the concrete background of anandamide and palmitoylethanolamide interference with bronchopulmonary inflammatory response in mice needs further clarification. In conclusion, we have demonstrated that different agonists of cannabinoid receptors can downmodulate bronchopulmonary inflammation in mice and suggest that this effect of cannabinoid receptor agonists could be mediated at least in part by cannabinoid CBz receptor. Acknowledgement manuscript.
1. 2. 3.
: The authors
than G. Bouer for his assistance
in preparing
the
References BERDYCHEV, EV., E. BOICHOT and V. LAGENTE J. Lipid Mediators Cell Signalling. 15 49-67 (1996). KAMINSKI NE. Biochem. Pharmacol. 52 1133-l 140 (1996). KLEIN TW., C. NEWTON, W. ZHU, Y. DAAKA and H. FRIEDMAN. Proc. Sot. Exp. Biol. Med. 209 205-212 (1995).
Vol. 63,No. 8,1!998
4. 5. 6.
7. 8. 9. 10.
CamubmoidLigandsand PulmonaryInflammation
PL129
MORAHAN, PS., PC. KLYKKEN, SH. SMITH, LS. HARRIS and AE. MUNSON. Infect. Immun. 23 670-674 (1979). FELDER CC., KE. JOYCE, EM. BRILEY, J. MANSOURI, K. MACKIE, 0. BLOND, Y. LAI, AL. MA and RL. MITCHELL. Mol. Pharmacol. 48 443-450 (1995). GALIEGUE S., S. MARY, J. MARCHAND, D. DUSSOSSOY, D. CARRIERE, P. CARAYON, M. BOUABALA, D. Shire, G. LEFUR and P. CASSELLAS. Eur. J. Biochem. 232 54-61 (1995). FISCHER-STENGER K., DAD. PETTIT and GA. CABRAL. J. Pharmacol. Exp. Ther. 267 1558-1565 (1993). BERDYCHEV EV., E. BOICHOT, N. GERMAIN, N. ALLAIN, JP. ANGER and V. LAGENTE Eur. J. Pharmacol. 330 231-240 (1997). STARK, S., MA. PACHECO and SR. CHILDERS. Cell. Mol. Neurobiol. 17 483-493 (1997). FACCI L., R. DAL TOSO, S. ROMANELLO, A. BURIANI, SD. SKAPER and A. LEON. Proc. Natl. Acad. Sci. USA 92 3376-3380 (1995).