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Involvement of RelB, a member of the NF-κB family, in morphine-induced immunosuppression in mouse
BRAIN, BEHAVIOR, and IMMUNITY Brain, Behavior, and Immunity 20 (2006) e45–e57 www.elsevier.com/locate/ybrbi
PNIRS Meeting Abstracts
PNIRS 2006 Abstracts M to P Stimulated production of interleukin-8 covaries with psychosocial risk factors for inflammatory disease among middle-aged community volunteers Anna L. Marsland a, Ramasri Sathanoori a, Matthew F. Muldoon b, Stephen B. Manuck a a b
University of Pittsburgh, USA University of Pittsburgh, School of Medicine, USA
A growing literature suggests that psychosocial factors are associated with increased vulnerability to inflammatory disease; however, the mechanisms of this effect remain unclear. Recent attention has focused on the possibility that psychosocial characteristics are associated with activation of innate immune pathways implicated in the pathogenesis of inflammatory disease. In this regard, chronic stress and depression have been associated with immune changes that are consistent with activation of inflammatory pathways. The goal of the current study was to extend these findings by exploring a broader range of psychosocial risk factors for inflammatory disease and a measure of inflammatory potential, rather than of current immune activation. For this purpose, we examined the relation of symptoms of depression, trait negative affect, perceived stress, and social support to lipopolysaccharide-induced production of the monocyte-derived pro-inflammatory cytokines/chemokines interleukin (IL)- 1b, IL-6, TNF-a, and IL-8 among a community sample of 183 healthy adults aged 30–54 years (mean = 45). Participants (59% male; 92% Caucasian, 7% African-American) were enrolled in the Adult Health and Behavior (AHAB) project, an ongoing study of cardiovascular risk. After controlling for age, gender, race, years of education, body mass index, current smoking, alcohol use, blood pressure, and white blood cell count, higher stimulated production of IL-8 was positively associated with symptoms of depression (b = .16, p < .04), trait negative affect (b = .18, p < .01), and perceived stress (b = .14, p < .04). In contrast, there was an inverse relationship between stimulated levels of IL-8 and trait conscientiousness (b = .15, p < .04) and perceived social support (b = .17, p < .02). The relationship between negative affect measures and IL-8 was independent of social support. Although there were sig-
doi:10.1016/j.bbi.2006.04.085
nificant univariate associations between higher IL-6 production and symptoms of depression and less social support, these relationships did not withstand adjustment for demographic controls. There were no significant associations between IL-1b or TNF-a and any of the psychosocial parameters. Our findings suggest that individuals at greater psychosocial risk for the development of inflammatory diseases, including cardiovascular disease, also show greater stimulated production of the pro-inflammatory chemokine, IL-8. Further exploration of this potential psychophysiological pathway is warranted. doi:10.1016/j.bbi.2006.04.086
Involvement of RelB, a member of the NF-jB family, in morphine-induced immunosuppression in mouse Cataldo Martucci, Silvia Franchi, Alberto E. Panerai, Paola Sacerdote Department of Pharmacology, University of Milano, Milano, Italy Opioid drugs remain the mainstay treatment for acute and chronic pain conditions beside still being common drugs of abuse. In vivo administration of the opioid drug morphine induces a decrease of multiple immune parameters, affecting almost all cells of innate and acquired immunity. The mechanisms involved in immunosuppression have not been well defined yet and its intracellular molecular targets in immune cells are far from being elucidated. The transcription factor NF-jB is a central regulator of immunity and of the NF-jB family; RelB is particularly involved in the expression of genes important in immune responses. We investigated the involvement of RelB in morphine-induced immunosuppression in mice deficient for the RelB factor. RelB / mice and wild type controls were injected s.c. with morphine 20 mg/Kg, and 1 h later immune parameters evaluated. Peritoneal macrophages were collected and cultured in vitro in the presence and in the absence of LPS. Morphine significantly reduced both spontaneous and LPS-stimulated macrophage production of the proinflammatory cytokines IL1b, TNFa, and IL12
e46
Abstracts / Brain, Behavior, and Immunity 20 (2006) e45–e57
in WT animals, while the drug failed to diminish these cytokines’ production in the RelB / mice. In contrast, the anti-inflammatory cytokine IL10 was similarly affected in the two strains. We also tested the ability of morphine to inhibit a fundamental and basic macrophage function, chemotaxis, in relB / animals. Using the classical Boyden method, we observed that morphine similarly decreased macrophage chemotaxis in the presence or in the absence of RelB, thus suggesting that this transcriptional factor is not necessary for the precocious steps of activation of macrophages. We then evaluated splenocyte Th1 and Th2 cytokines production both spontaneous and stimulated with Con-A. We observed a clear morphine-induced reduction of IL2 and IFNc production by WT splenocytes, whereas no effect of the drug was observed in RelB / mice. On the contrary the production of the Th2 cytokines IL4 and IL10 was lessened to the same degree by morphine in wild type and RelB / mice. Finally, a FACS analysis of the peritoneal and splenocyte population showed that neither morphine nor the lack of RelB affected the number and percentage of cells. In conclusion, our data suggest that RelB is a necessary and sufficient target for the modulation by morphine of pro-inflammatory and Th1 cytokines, but not for anti-inflammatory and Th2 cytokines. They also indicate that morphine utilizes multiple intracellular pathways in order to exert its generalized immunosuppression. doi:10.1016/j.bbi.2006.04.087
Stress-impaired mucosal wound healing in humans involves an early overexpression of chemokines and E-selectin Phillip T. Marucha, Zong Juan Fong, Christopher G. Engeland Department of Periodontics, University of Illinois at Chicago, Chicago, IL, USA Psychosocial stress has been shown to impair wound healing in human and animal models. This is presumed to be due to dysregulation of inflammatory responses required for wound repair. Mucosal wounds heal rapidly with minimal inflammation and relatively little scarring. We have demonstrated that mucosal wounds under stress conditions heal slower with increased expression of proinflammatory cytokines. In this study, we sought to determine whether stress altered the expression of chemokines and adhesion molecules important in cell recruitment and activation during mucosal wound healing. 28 males and 15 females had experimental wounds placed during a stress period (i.e., academic examinations) and a nonstress period (summer vacation). Two wounds were placed on the hard palates at each time: a 3.5 mm circular wound and a 1 · 5 mm longitudinal wound. The 3.5 mm wound was videographed daily and measured
by photoplanimetry. The 1 · 5 mm wound was biopsied at 6 or 24 h later so that each person received their biopsy at the same interval during the stress and nonstress time. During the examination time, individuals reported significantly more perceived stress, state anxiety and tension (p < .001), than the same students during summer vacation. Students had significantly larger wounds 24 h after wounding during the stress period (p < .05). Previously unwounded tissue and biposied tissue was analyzed by real time PCR for IL-8, MIP-1a, MCP-1, E-selectin, and ICAM. In previously unwounded tissue, E-selectin (P < .001, >2-fold increase) and MIP-1a (P < .01, >2-fold increase) were significantly higher during the stress compared to the nonstress condition. Biopsies harvested 6 h after wounding showed significantly larger increases in E-selectin (P < .009, >3-fold increase), MIP-1a (P < .02, >2-fold increase), and IL-8 (P < .03, >50% increase), while ICAM approached significance (P < .09, >80% increase) during the stress condition as compared to the nonstress. There were no significant stress-alone differences in expression of the target genes in biopsies one day after wounding. Interestingly, females had higher levels of E-selectin (P < .05, 2-fold increase), while males did not, 24 h after wounding. Furthermore, previous data has also demonstrated that females close mucosal wounds slower than males. These data demonstrate that chemokines important in neutrophil (IL-8) and macrophage (MIP-1a) recruitment, and specific adhesion molecules are increased during stress. Previous studies have demonstrated that over-recruitment of neutrophils and macrophages can lead to enhanced production of pro-inflammatory cytokines, impaired healing, and scar formation in mucosa. Thus, along with cytokine expression, enhanced cell recruitment appears to be important in the modulation of mucosal wound healing during stress. Acknowledgments Supported by NIH/NIDCR RO1DE12792. doi:10.1016/j.bbi.2006.04.088
Epigenetic effects of psychosocial distress on peripheral blood leukocytes of women with breast cancer Herbert L. Mathews, Linda W. Janusek Loyola University of Chicago, USA Women diagnosed with breast cancer experience psychosocial distress that is characterized by elevated perceived stress, mood disturbance and anxiety. Accompanying these forms of psychosocial distress is immune dysregulation. The overall purpose of this study was to determine whether peripheral blood mononuclear cell (PBMC) epigenetic modifications, subsequent to psycho-
PNIRS Meeting Abstracts
PNIRS 2006 Abstracts M to P Stimulated production of interleukin-8 covaries with psychosocial risk factors for inflammatory disease among middle-aged community volunteers Anna L. Marsland a, Ramasri Sathanoori a, Matthew F. Muldoon b, Stephen B. Manuck a a b
University of Pittsburgh, USA University of Pittsburgh, School of Medicine, USA
A growing literature suggests that psychosocial factors are associated with increased vulnerability to inflammatory disease; however, the mechanisms of this effect remain unclear. Recent attention has focused on the possibility that psychosocial characteristics are associated with activation of innate immune pathways implicated in the pathogenesis of inflammatory disease. In this regard, chronic stress and depression have been associated with immune changes that are consistent with activation of inflammatory pathways. The goal of the current study was to extend these findings by exploring a broader range of psychosocial risk factors for inflammatory disease and a measure of inflammatory potential, rather than of current immune activation. For this purpose, we examined the relation of symptoms of depression, trait negative affect, perceived stress, and social support to lipopolysaccharide-induced production of the monocyte-derived pro-inflammatory cytokines/chemokines interleukin (IL)- 1b, IL-6, TNF-a, and IL-8 among a community sample of 183 healthy adults aged 30–54 years (mean = 45). Participants (59% male; 92% Caucasian, 7% African-American) were enrolled in the Adult Health and Behavior (AHAB) project, an ongoing study of cardiovascular risk. After controlling for age, gender, race, years of education, body mass index, current smoking, alcohol use, blood pressure, and white blood cell count, higher stimulated production of IL-8 was positively associated with symptoms of depression (b = .16, p < .04), trait negative affect (b = .18, p < .01), and perceived stress (b = .14, p < .04). In contrast, there was an inverse relationship between stimulated levels of IL-8 and trait conscientiousness (b = .15, p < .04) and perceived social support (b = .17, p < .02). The relationship between negative affect measures and IL-8 was independent of social support. Although there were sig-
doi:10.1016/j.bbi.2006.04.085
nificant univariate associations between higher IL-6 production and symptoms of depression and less social support, these relationships did not withstand adjustment for demographic controls. There were no significant associations between IL-1b or TNF-a and any of the psychosocial parameters. Our findings suggest that individuals at greater psychosocial risk for the development of inflammatory diseases, including cardiovascular disease, also show greater stimulated production of the pro-inflammatory chemokine, IL-8. Further exploration of this potential psychophysiological pathway is warranted. doi:10.1016/j.bbi.2006.04.086
Involvement of RelB, a member of the NF-jB family, in morphine-induced immunosuppression in mouse Cataldo Martucci, Silvia Franchi, Alberto E. Panerai, Paola Sacerdote Department of Pharmacology, University of Milano, Milano, Italy Opioid drugs remain the mainstay treatment for acute and chronic pain conditions beside still being common drugs of abuse. In vivo administration of the opioid drug morphine induces a decrease of multiple immune parameters, affecting almost all cells of innate and acquired immunity. The mechanisms involved in immunosuppression have not been well defined yet and its intracellular molecular targets in immune cells are far from being elucidated. The transcription factor NF-jB is a central regulator of immunity and of the NF-jB family; RelB is particularly involved in the expression of genes important in immune responses. We investigated the involvement of RelB in morphine-induced immunosuppression in mice deficient for the RelB factor. RelB / mice and wild type controls were injected s.c. with morphine 20 mg/Kg, and 1 h later immune parameters evaluated. Peritoneal macrophages were collected and cultured in vitro in the presence and in the absence of LPS. Morphine significantly reduced both spontaneous and LPS-stimulated macrophage production of the proinflammatory cytokines IL1b, TNFa, and IL12
e46
Abstracts / Brain, Behavior, and Immunity 20 (2006) e45–e57
in WT animals, while the drug failed to diminish these cytokines’ production in the RelB / mice. In contrast, the anti-inflammatory cytokine IL10 was similarly affected in the two strains. We also tested the ability of morphine to inhibit a fundamental and basic macrophage function, chemotaxis, in relB / animals. Using the classical Boyden method, we observed that morphine similarly decreased macrophage chemotaxis in the presence or in the absence of RelB, thus suggesting that this transcriptional factor is not necessary for the precocious steps of activation of macrophages. We then evaluated splenocyte Th1 and Th2 cytokines production both spontaneous and stimulated with Con-A. We observed a clear morphine-induced reduction of IL2 and IFNc production by WT splenocytes, whereas no effect of the drug was observed in RelB / mice. On the contrary the production of the Th2 cytokines IL4 and IL10 was lessened to the same degree by morphine in wild type and RelB / mice. Finally, a FACS analysis of the peritoneal and splenocyte population showed that neither morphine nor the lack of RelB affected the number and percentage of cells. In conclusion, our data suggest that RelB is a necessary and sufficient target for the modulation by morphine of pro-inflammatory and Th1 cytokines, but not for anti-inflammatory and Th2 cytokines. They also indicate that morphine utilizes multiple intracellular pathways in order to exert its generalized immunosuppression. doi:10.1016/j.bbi.2006.04.087
Stress-impaired mucosal wound healing in humans involves an early overexpression of chemokines and E-selectin Phillip T. Marucha, Zong Juan Fong, Christopher G. Engeland Department of Periodontics, University of Illinois at Chicago, Chicago, IL, USA Psychosocial stress has been shown to impair wound healing in human and animal models. This is presumed to be due to dysregulation of inflammatory responses required for wound repair. Mucosal wounds heal rapidly with minimal inflammation and relatively little scarring. We have demonstrated that mucosal wounds under stress conditions heal slower with increased expression of proinflammatory cytokines. In this study, we sought to determine whether stress altered the expression of chemokines and adhesion molecules important in cell recruitment and activation during mucosal wound healing. 28 males and 15 females had experimental wounds placed during a stress period (i.e., academic examinations) and a nonstress period (summer vacation). Two wounds were placed on the hard palates at each time: a 3.5 mm circular wound and a 1 · 5 mm longitudinal wound. The 3.5 mm wound was videographed daily and measured
by photoplanimetry. The 1 · 5 mm wound was biopsied at 6 or 24 h later so that each person received their biopsy at the same interval during the stress and nonstress time. During the examination time, individuals reported significantly more perceived stress, state anxiety and tension (p < .001), than the same students during summer vacation. Students had significantly larger wounds 24 h after wounding during the stress period (p < .05). Previously unwounded tissue and biposied tissue was analyzed by real time PCR for IL-8, MIP-1a, MCP-1, E-selectin, and ICAM. In previously unwounded tissue, E-selectin (P < .001, >2-fold increase) and MIP-1a (P < .01, >2-fold increase) were significantly higher during the stress compared to the nonstress condition. Biopsies harvested 6 h after wounding showed significantly larger increases in E-selectin (P < .009, >3-fold increase), MIP-1a (P < .02, >2-fold increase), and IL-8 (P < .03, >50% increase), while ICAM approached significance (P < .09, >80% increase) during the stress condition as compared to the nonstress. There were no significant stress-alone differences in expression of the target genes in biopsies one day after wounding. Interestingly, females had higher levels of E-selectin (P < .05, 2-fold increase), while males did not, 24 h after wounding. Furthermore, previous data has also demonstrated that females close mucosal wounds slower than males. These data demonstrate that chemokines important in neutrophil (IL-8) and macrophage (MIP-1a) recruitment, and specific adhesion molecules are increased during stress. Previous studies have demonstrated that over-recruitment of neutrophils and macrophages can lead to enhanced production of pro-inflammatory cytokines, impaired healing, and scar formation in mucosa. Thus, along with cytokine expression, enhanced cell recruitment appears to be important in the modulation of mucosal wound healing during stress. Acknowledgments Supported by NIH/NIDCR RO1DE12792. doi:10.1016/j.bbi.2006.04.088
Epigenetic effects of psychosocial distress on peripheral blood leukocytes of women with breast cancer Herbert L. Mathews, Linda W. Janusek Loyola University of Chicago, USA Women diagnosed with breast cancer experience psychosocial distress that is characterized by elevated perceived stress, mood disturbance and anxiety. Accompanying these forms of psychosocial distress is immune dysregulation. The overall purpose of this study was to determine whether peripheral blood mononuclear cell (PBMC) epigenetic modifications, subsequent to psycho-