Beta adrenoceptor regulation of macrophage arginase activity.

BACKGROUND: Arginase, which metabolizes L-arginine within the urea cycle, is essential for production of polyamines and affects production of nitric oxide by depletion of L-arginine, the common substrate for both arginase and nitric oxide synthase. Having shown that trauma increases splenic macrophage arginase activity, we seek to define the mechanisms for this. RAW macrophage arginase activity and expression are increased by 8-bromo-cAMP in vitro. We hypothesize that since catecholamines increase cAMP, trauma-induced splenic arginase activity may be mediated by post-injury catecholamine release. METHODS: RAW 264.7 macrophage arginase activity was measured in vitro in response to 4 catecholamines with or without propranolol or lipopolysaccharide (LPS). C57BL/6 mice underwent laparotomy as a model of moderate trauma after propranolol treatment, with and without intraperitoneal Escherichia coli LPS administration as a simulated pro-inflammatory stimulus. RESULTS: Macrophage arginase activity increased in vitro in response to catecholamines or LPS (P < .05). Propranolol pretreatment blocked macrophage arginase activity induced by epinephrine (10 mumol/L) in vitro (P < .05). Trauma or LPS alone increased splenic arginase activity in vivo (P < .05). Propranolol did not alter LPS-induced splenic arginase activity but did significantly reduce trauma-induced splenic arginase activity (P < .05). CONCLUSIONS: Catecholamines alone increase macrophage arginase activity through beta-adrenoceptor activation. Increased splenic arginase activity induced by moderate trauma is decreased by beta-adrenoceptor blockade, suggesting that trauma-induced arginase activity is partly mediated by endogenous catecholamines.

Anxiety in parents of young febrile children in a pediatric emergency department: why is it elevated?

This study assessed anxiety levels of parents of young febrile children who presented to a pediatric emergency department (ED) with fever. One hundred and seventy parents completed a 90-item questionnaire. Anxiety was measured by use of the State Trait Anxiety Inventory. Parents were asked what they had previously thought about and how they felt about the ED process. Mean parental anxiety was 50.1 (95% CI 48.1, 52.2), significantly elevated from adult standards (p < 0.0001). A multivariate model comprising: (1) feeling "not at all" well rested, (2) having no other children, (3) having thought about a blood test, and (4) feeling worried about trusting the physician was associated with elevated anxiety. In conclusion, parents of febrile young children in the ED are very anxious.

Role of virus replication in a murine model of AIDS-associated interstitial pneumonitis.

One of the major complications of HIV infection is the development of interstitial pneumonitis (IP). IP is characterized by lymphocytic infiltration of the lung and may lead to respiratory failure in some cases. The etiology of IP is unknown although it is likely the result of an antiviral or autoimmune response occurring in the lung. To determine the role of viral replication in the development of IP, AZT was evaluated for the ability to inhibit development of lung inflammation in a murine model of retrovirus-associated IP. Mice were infected with LP-BM5 retrovirus, which induces murine AIDS. Infected mice develop IP by 4 weeks postinfection characterized by infiltration of the lung with activated T cells, B cells, and macrophages. Virus could be detected in the lungs of these mice by 2 weeks postinfection and persisted throughout the course of disease. To determine if reduction in viral load affected the disease process, infected mice were treated with AZT for varying periods postinfection and analyzed for the development of IP. Treatment with AZT resulted in a treatment time-dependent reduction of viral RNA in the lungs of infected mice compared to untreated infected mice. The reduction of viral burden in the lungs correlated with a reduction in the severity of IP and decreased production of the proinflammatory cytokines interleukin (IL)-1 beta and interferon (IFN)-gamma. These results suggest that continuous viral replication in the lung contributes to the pathogenesis of IP.

Role of T cell subsets in the development of AIDS-associated interstitial pneumonitis in mice.

Idiopathic interstitial pneumonitis (IP), characterized by lymphocytic infiltration of the lung and pulmonary dysfunction, is a major noninfectious complication of human immunodeficiency virus (HIV) infection. The role of the CD4+ and CD8+ T cell populations and INF-gamma in the development of IP were analyzed using a murine model of retroviral-associated IP. Infected mice depleted of CD8+ T cells developed IP similarly to untreated infected mice, suggesting that the CD8+ T cell population does not play a role in IP. Furthermore, depletion of CD8+ T cells did not alter the level of viral RNA in lungs, suggesting that cytotoxic T cells may not serve a role in controlling virus burden in lungs. In contrast, depletion of CD4+ T cells in infected mice prevented the development of IP and inhibited inflammatory cytokine expression, suggesting that CD4+ T cells are important for the development of IP. IFN-gamma -/- mice infected with virus for 10 weeks developed IP, although the severity of lymphocytic infiltration was substantially reduced compared to infected wild-type mice. The data suggest that persistent viral antigen in the lung may drive a CD4+ T cell-mediated immune response, resulting in the chronic production of IFN-gamma which amplifies a chronic inflammatory response in the lung resulting in tissue injury.

Pulmonary lymphoid cell activation and cytokine expression in murine AIDS-associated interstitial pneumonitis.

Limited information is available about the pathogenesis of acquired immune deficiency syndrome (AIDS)-associated idiopathic interstitial pneumonitis, a common noninfectious complication of human immunodeficiency virus (HIV) infection. Infection of C57B1/6 mice with LP-BM5 retrovirus, a murine model of AIDS, leads to development of a diffuse interstitial pneumonitis that displays many features of human AIDS-associated interstitial pneumonitis. To further characterize the cellular and molecular features of this lung disease, the temporal development of cellular infiltration, cytokine expression, and virus replication were evaluated in lung tissue of virus-infected mice. Persistent expression of viral RNA was detectable in lungs as early as 1 wk after infection. Infiltration of the lungs by CD4+ and CD8+ T cells, by IgG+ and IgA+ B cells, and by macrophages was observed by 4 wk after infection and continued through 8 wk of infection. Histologically, cellular infiltration was most pronounced in peribronchial and perivascular regions, whereas inflammation of alveolar septae and alveolar spaces was minimal. In contrast to normals, T cells from infected lungs were immunodeficient in that they failed to proliferate in response to the mitogen concanavalin A (ConA). However, evaluation of cytokine mRNA expression by interstitial lung lymphoid cells indicated that cells from infected lungs were chronically activated, in that elevated expression of interferon-gamma (IFN-gamma) and interleukin-10 (IL-10) was observed throughout the course of infection. Similarly, expression by interstitial lung lymphoid cells of mRNA for the proinflammatory cytokine IL-1 and the fibrogenic cytokine transforming growth factor-beta (TGF-beta) was also increased following infection. These results indicate that retrovirus-induced immunodeficiency in mice is associated with infiltration and chronic activation of lymphoid cells in the lungs. Furthermore, simultaneous expression of IL-10, IFN-gamma, and TGF-beta suggests that cytokine-expressing cells in infected lungs may be unresponsive to inhibitory and antiinflammatory effects of IL-10 and/or TGF-beta, thus contributing to chronicity of inflammation in this disorder.

Defective CD4+ T cell signaling in murine AIDS: uncoupling of the T cell receptor complex from PIP2 hydrolysis.

CD4+ T cells from mice with murine AIDS (MAIDS) have been shown to be unable to respond to TCR stimulation as measured by proliferation, IL-2 production, or IL-2R upregulation, although responsiveness was restored with PMA and ionomycin. In this report we have demonstrated that the inability of MAIDS CD4+ T cells to respond to CD3 stimulation was not associated with reduced surface expression of CD3, CD4, or CD28 and could not be overcome by costimulation with anti-CD28 antibody. However, MAIDS CD4+ T cells failed to activate the PIP2 hydrolysis pathway efficiently, resulting in diminished IP3 production and reduced Ca2+ mobilization compared to normal controls. Additionally, TCR signaling in MAIDS resulted in a reduction in the level of tyrosine phosphorylation of some proteins including deficient tyrosine phosphorylation of PLC-gamma 1, compared to normal CD4+ T cells. These studies suggest that stimulation through the TCR in CD4+ T cells from MAIDS-infected mice is uncoupled from the phosphotidylinositol hydrolysis pathway due to deficient activation of PLC-gamma 1.

Ethanol as a possible cofactor in the development of murine AIDS.

Chronic ethanol (EtOH) abuse in humans leads to a variety of immunomodulatory events that can alter resistance to a number of infectious agents. Whether alcohol abuse affects the susceptibility to human immunodeficiency virus infection or the subsequent development of acquired immune deficiency syndrome (AIDS) is a matter of extreme importance; however, available information in humans or animal models is limited. The goal of this study was to evaluate the effect of chronic EtOH feeding in mice on the development of immunodeficiency in the murine model of AIDS (MAIDS). C57BI/6 mice were placed on the Lieber-DeCarli liquid EtOH diet (25% or 31% total caloric intake) or a nutrient-matched isocaloric liquid control diet. Seven days later, mice were infected with the LP-BM5 murine leukemia virus mixture, and groups of infected and noninfected mice were assayed at defined time points postinfection for antigen-specific and nonspecific immune responses. In the absence of retroviral infection, chronic EtOH feeding (5-8 weeks) led to reductions in spleen weights, compared with isocaloric controls. In spite of reduced spleen size, mitogenic responses of spleen cells to concanavalin A (ConA) and lipopolysaccharide (LPS) were elevated in EtOH-fed mice, as compared with mice fed the control diet. Chronic EtOH feeding also enhanced the allogeneic mixed lymphocyte response and increased antigen-specific priming of both B-cells and CD4+ T-cells to the antigen, sheep red blood cells. In MAIDS-infected mice, chronic EtOH feeding delayed but did not prevent the onset of virus-induced immunodeficiency and MAIDS-induced autoantibody synthesis.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of chronic alcohol feeding and murine AIDS virus infection on liver antioxidant defense systems in mice.

Whether ethanol (ETOH) abuse could contribute to the development of acquired immunodeficiency syndrome (AIDS) among human immunodeficiency virus (HIV)-positive drug abusers is a critical question for which little experimental information is available. This study was designed to determine if chronic ETOH feeding and murine AIDS virus infection cooperatively affected liver antioxidant defense systems in C57B1/6 female mice. Mice were divided into two groups and fed the Lieber-DeCarli liquid ETOH diet containing ETOH at a concentration to provide 31% of total caloric intake or an isocaloric liquid control (control) diet in which dextrin-maltose replaced ETOH. One week after the initiation of ETOH feeding, half of the mice in each diet group (8 mice) were injected intraperitoneally with murine retrovirus (MAIDS) stock. After 3 and 5 weeks of ETOH feeding, half of the mice in each of the four treatment groups (4 mice) were killed, and livers were excised for biochemical analysis. Liver reduced glutathione (GSH) levels and activities of glutathione peroxidase (GP), glutathione reductase (GR), glutathione transferase (GT), catalase and superoxide dismutase (SOD), and serum ETOH concentrations were determined. The results demonstrated that serum ETOH concentrations were significantly elevated in ETOH-MAIDS group when compared with the ETOH group. Moreover, chronic ETOH feeding and MAIDS infection independently depressed liver antioxidant defense capability, and together led to an additive inhibition of GSH and SOD activities. In addition, MAIDS infection inhibited an ETOH-induced increase in catalase and GT activities. These results suggest that alcohol abuse could contribute to the development of AIDS by inhibiting the protective capability of an infected individual against oxidative stress.(ABSTRACT TRUNCATED AT 250 WORDS)

Activation-dependent apoptosis in CD4+ T cells during murine AIDS.

The mechanism by which CD4+ T cells are depleted during HIV infection remains a matter of controversy. Recent reports have suggested that activation-induced apoptosis of antigen-specific CD4+ T cells may lead ultimately to depletion of this T cell subset during HIV infection. The murine retroviral model of AIDS (MAIDS) also displays progressive immunodeficiency, but depletion of the CD4+ T cell subset is not characteristic of the disease. We report that a fraction of splenic CD4+ T cells from 8- to 14-week MAIDS-infected C57B1/6 mice, but not normal mice, was undergoing apoptosis at the time of cell isolation. Typical apoptotic morphology and internucleosomal DNA fragmentation was seen in CD4+ T cells only from infected mice. Moreover, injection of anti-CD3 mAb enhanced DNA fragmentation in CD4+ T cells from infected but not normal mice, suggesting that the apoptosis in vivo in CD4+ T cells during MAIDS may be dependent on cell activation. Induction of apoptosis was associated with defective signaling through the TcR complex, since anti-CD3 stimulation in vitro of CD4+ T cells from infected mice caused a diminished calcium response, yet no cellular proliferation. Despite the occurrence of apoptosis in vivo in CD4+ T cells from MAIDS-infected mice, CD4+ T cells were not depleted during the course of disease. Thus, while apoptosis in CD4+ T cells is a characteristic of MAIDS immunodeficiency disease as well as HIV infections in humans, CD4+ T cell depletion is only observed in HIV infections. In view of the extensive lymphocyte expansion which occurs in vivo in MAIDS, the balance between activation-induced apoptosis and chronic cell proliferation may determine whether cell depletion is a characteristic feature of retrovirus-induced immunodeficiencies.

T-deficient transmembrane signaling in CD4+ T cells of retroviral-induced immune-deficient mice.

The defective virus found in the LP-BM5 mixture of murine leukemia viruses induces a severe immune deficiency disease in C57BL/6 mice that is characterized by the activation and expansion of T and B cells that become unresponsive to normal immune stimuli. The nature of the biochemical lesion in these defective lymphocyte populations remains unknown. Flow cytometric analysis of the T cell population in infected animals has demonstrated expansion of both CD4+ and CD8+ subsets. Despite chronic expansion in vivo, CD4+ T cells by wk 4 postinfection failed to up-regulate cell surface IL-2R expression, produced IL-2, or proliferate in vitro in response to either Con A, Staphylococcal enterotoxin super-antigens, or anti-CD3 stimulation. Exogenous IL-2 did not restore the proliferative response and also failed to up-regulate IL-R expression on CD4+ T cells from infected mice, even though basal IL-2R expression was initially elevated compared to normals. In contrast, CD4+ T cells from infected mice could be induced to proliferate by stimulation with PMA and ionomycin resulting in IL-2R up-regulation, IL-2 production, and proliferation. Moreover, proliferation could also be induced by anti-CD3 plus PMA, although anti-CD3 plus ionomycin was without effect. These studies suggest that chronic expansion of CD4+ T cells in infected mice is probably not maintained by normal TCR signaling, which appears defective in these cells. In addition, the lesion in biochemical signaling appears to result in defective activation of protein kinase C, which can be overcome by direct activation with PMA.

Changes in the carbon and nitrogen status of forest soil organic horizons between 1949/50 and 1987.

Forest soil organic horizons from 15 profiles in NE Scotland originally sampled in 1949/50, were resampled in 1987. Analyses of both sets of soils for organic C and N show that although concentrations of the two elements have decreased with time, there has been a large increase in storage due to an increase in O horizon thickness. In most cases surface organic horizons have become more acid between 1949/50 and 1987. Calculated mean accumulation rates for C and N are 353.4 kg ha(-1) year(-1) and 21.2 kg ha(-1) year(-1) respectively. Changes in the C/N ratio with time give no indication of progressive N saturation and suggest sudden breakthrough of N in drainage water is not imminent.

Dietary self-selection and the Zucker rat.

Impaired protein deposition has been suggested to be a critical factor promoting the hyperphagia of the obese Zucker rat. The selection patterns of adult, male, genetically obese (fa/fa) and lean (Fa/-) Zucker rats were studied to determine if obese rats would select a diet that was higher in protein than the diet selected by lean littermates. Rats were provided ad libitum access to three macronutrient sources and were allowed to compose their own diets for 9 days. The three dietary items were: a vitamin + mineral--supplemented carbohydrate source (cornstarch), a vitamin + mineral--supplemented protein source (casein) and commercially available corn oil. Obese rats ate 43% more calories than lean littermates. Further, obese rats selected a diet that provided 12% of their total caloric intake as protein, 24% as carbohydrate and 64% as fat. Lean rats selected a diet that provided 33% of their total caloric intake as protein, 37% as carbohydrate and 30% as fat. These selection data are not consistent with the hypothesized importance of the role of dietary protein and its incorporation into lean body mass as a stimulus promoting the hyperphagia demonstrated by the genetically obese Zucker rats.