Parasites, nutrition, immune responses and biology of metabolic tissues.

Nutritional immunology, immunometabolism and identification of novel immunotherapeutic targets are areas of active investigation in parasitology. There is a well-documented crosstalk among immune cells and cells in metabolically active tissues that is important for homeostasis. The numbers and function of these cells are altered by obesity leading to inflammation. A variety of helminths spend some part of their life cycle in the gastrointestinal tract and even entirely enteral nematode infections exert beneficial effects on glucose and lipid metabolism. The foundation of this review is the ability of enteric nematode infections to improve obesity-induced type 2 diabetes and the metabolic syndrome, which are significant health issues in developed areas. It considers the impact of nutrition and specific nutritional deficiencies, which are occur in both undeveloped and developed areas, on the host's ability mount a protective immune response against parasitic nematodes. There are a number of proposed mechanisms by which parasitic nematodes can impact metabolism including effects gastrointestinal hormones, altering epithelial function and changing the number and/or phenotype of immune cells in metabolic tissues. Nematodes can also exert their beneficial effects through Th2 cytokines that activate the transcription factor STAT6, which upregulates genes that regulate glucose and lipid metabolism.

An unsuspected cause of diarrhoea and gastrointestinal bleeding during corticosteroid therapy.

We report on a fatal case of disseminated strongyloidiasis during corticosteroid treatment presenting with abdominal pain, diarrhoea and lower gastrointestinal bleeding. The patient emigrated from Thailand 16 years before the current hospitalisation. Complicated strongyloidiasis is a relatively unrecognized complication of corticosteroid therapy in non-endemic areas. In individuals who have resided in endemic areas, even decades before treatment, strongyloidiasis should be excluded before initiation of immunosuppressants.

Mechanisms of smooth muscle responses to inflammation.

BACKGROUND: Inflammation-induced changes in smooth muscle may be the consequence of changes in the properties of smooth muscle itself, in the control by nerves and hormones, in the microenvironment, or in the balance of constitutive or induced mediators. A general concept is that the specific characteristics and effects of inflammation can be linked to the nature of the infiltrate and the associated mediators, which are dictated predominantly by the immune environment. Inflammatory mediators may regulate smooth muscle function by directly acting on smooth muscle cells or, indirectly, through stimulation of the release of mediators from other cells. In addition, smooth muscle is not a passive bystander during inflammation and our knowledge of molecular signaling pathways that control smooth muscle function, and the contribution of the immune mechanisms to smooth muscle homeostasis, has expanded greatly in the last decade. Recent studies also demonstrated the relevance of extracellular proteases, of endogenous or exogenous origin, redox imbalance, or epigenetic mechanisms, to gastrointestinal dismotility and inflammation in the context of functional and organic disorders. PURPOSE: In this review we discuss the various types of inflammation and the established and emerging mechansims of inflammation-induced changes in smooth muscle morphology and function.

Role of enteric nerves in immune-mediated changes in protease-activated receptor 2 effects on gut function.

BACKGROUND: Protease-activated receptors (PARs) are expressed on structural and immune cells. Control of initiation, duration, and magnitude of PAR effects is linked to the level of receptor expression, availability of proteases, and the intracellular signal transduction machinery. We investigated nematode infection-induced changes in PAR(2) expression and the impact on smooth muscle and epithelial responses to PAR(2) agonists. METHODS: Smooth muscle and epithelial cell function were assessed in wild-type, and IL-4, IL-13 or STAT6 gene-deficient mice following treatment with vehicle, Nippostrongylus brasiliensis or Heligmosomoides polygyrus, or IL-13. The role of enteric nerves was determined using tetrodotoxin to block nerve conduction. Expression of PAR(2) was assessed by real-time PCR, western blot and immunohistochemistry. KEY RESULTS: Nematode infection induced a STAT6- and IL-13-dependent up-regulation of PAR(2) mRNA expression. The infection-induced hypercontractility to PAR(2) agonists required STAT6/IL-13 and was neurally mediated. In contrast, the infection-induced decrease in epithelial secretion to PAR(2) agonists was partly dependent on STAT6 and independent of enteric nerves. The hyposecretion was correlated with decreased PAR(2) immunofluorescent staining on the apical surface of epithelial cells, but enhanced lamina propria immunostaining for PAR(2). CONCLUSIONS & INFERENCES: This is the first study to demonstrate an immune regulation of PAR(2) expression that impacts both smooth muscle and epithelial cell responses to PAR(2) agonists. Differences in responses between smooth muscle and epithelial cells are related to the contribution of enteric nerves. These data provide a mechanism by which activation of PAR(2) in immune-based pathologies can induce both transient and long-lasting changes in gut function.

Tumour necrosis factor alpha contributes to protection against Giardia lamblia infection in mice.

Giardia lamblia is a ubiquitous parasite that causes diarrhoea. Effective control of Giardia infections in mice has been shown to involve IgA, T cells, mast cells and IL-6. We now show that Tumour necrosis factor alpha (TNFalpha) also plays an important role in the early control of giardiasis. Mice treated with neutralizing anti-TNFalpha antibodies or genetically deficient in TNFalpha were infected with the G. lamblia clone GS/(M)-H7. In both cases, mice lacking TNFalpha had much higher parasite numbers than controls during the first 2 weeks of infections. However, anti-parasite IgA levels, mast cell responses, and IL-4 and IL-6 mRNA levels do not appear significantly altered in the absence of TNFalpha. In addition, we show that mice infected with G. lamblia exhibit increased intestinal permeability, similar to human Giardia infection, and that this increase occurs in both wild-type and TNFalpha deficient mice. We conclude that TNFalpha is essential for host resistance to G. lamblia infection, and that it does not exert its effects through mechanisms previously implicated in control of this parasite.

Changes in enteric neural circuitry and smooth muscle in the inflamed and infected gut.

Much of the morbidity associated with inflammatory bowel disease (IBD) and infection is caused by disordered gastrointestinal motor and secretory functions. Given that intestinal smooth muscle tone and epithelial cell secretion are regulated by the enteric nervous system (ENS), it is quite likely that inflammation-induced changes in the enteric neural circuitry contribute to intestinal dysmotility and diarrhoea. Indeed, discoveries over the past decades have demonstrated that gut inflammation and infections are associated with changes in key elements all along the enteric neural circuitry from the sensory transducers, the enterochromaffin (EC) cells, to the terminals of motor neurones.

AAVP Symposium: new approaches in the study of animal parasites.

The following three papers are a very small window onto the types of research being pursued by members of the American Association of Veterinary Parasitologists. They are related by the fact that newer areas in the biology of parasites and their hosts are discussed. The first paper by Dr. Tom Klei, gives a brief view of the interactions between host and parasite of the fascinating organism Wolbachia, a parasite of parasites. The second paper by Dr. Gloria Solano-Aguilar addresses the use of probiotics to alter the host­parasite interface and influence host resistance. The final paper by Dr. Lou Gasbarre outlines an example of integration of the genomics revolution into Veterinary Parasitology. While the subjects are diverse, they demonstrate the vitality of the AAVP.

Complement inhibitor, complement receptor 1-related gene/protein y-Ig attenuates intestinal damage after the onset of mesenteric ischemia/reperfusion injury in mice.

Complement receptor 1-related gene/protein y (Crry) is a murine membrane protein that regulates the activity of both classical and alternative complement pathways. We used a recombinant soluble form of Crry fused to the hinge, CH2, and CH3 domains of mouse IgG1 (Crry-Ig) to determine whether inhibition of complement activation prevents and/or reverses mesenteric ischemia/reperfusion-induced injury in mice. Mice were subjected to 30 min of ischemia, followed by 2 h of reperfusion. Crry-Ig was administered either 5 min before or 30 min after initiation of the reperfusion phase. Pretreatment with Crry-Ig reduced local intestinal mucosal injury and decreased generation of leukotriene B(4) (LTB(4)). When given 30 min after the beginning of the reperfusion phase, Crry-Ig resulted in a decrease in ischemia/reperfusion-induced intestinal mucosal injury comparable to that occurring when it was given 5 min before initiation of the reperfusion phase. The beneficial effect of Crry-Ig administered 30 min after the initiation of reperfusion coincided with a decrease in PGE(2) generation despite the fact that it did not prevent local infiltration of neutrophils and did not have a significant effect on LTB(4) production. These data suggest that complement inhibition protects animals from reperfusion-induced intestinal damage even if administered as late as 30 min into reperfusion and that the mechanism of protection is independent of neutrophil infiltration or LTB(4) inhibition.

Colitis-induced alterations in adrenergic control of circular smooth muscle in vitro in rats.

The present study investigated inflammation-induced changes in adrenergic regulation of smooth muscle. Colitis was induced in rats by intrarectal administration of trinitrobenzenesulfonic acid in ethanol. After 4 h (acute) or 7 days (chronic), in vitro isometric tension was measured in strips of circular smooth muscle taken from the distal colon. In controls, the major inhibitory control of smooth muscle responses to nerve stimulation was mediated by nitric oxide and beta adrenergic receptors. There was less evidence of alpha adrenergic control. Studies with the beta3 receptor antagonist cyanopindolol and the beta3 receptor agonist BRL37344 revealed that beta adrenergic regulation of spontaneous contractions and responses to nerve stimulation were mediated primarily by the beta3 adrenoreceptor. Both acute and chronic colitis significantly increased responses to electrical field stimulation. This effect was attributed to a loss of inhibitory nitrergic regulation as well as to selective changes in the beta adrenergic control of colonic circular smooth muscle. Inflammation did not alter alpha adrenergic control. Chronic colitis also decreased the sensitivity to nerve stimulation and pharmacological contractile agents. Acute and chronic inflammation reduced the ability of BRL37344 to inhibit contractions in response to nerve stimulation. In addition, in inflamed colon, BRL37344 was less effective in relaxing carbachol-induced precontractions. Finally, inflammation resulted in a loss of the ability of the cyanopindolol to increase the amplitude of both spontaneous contractions and contractions in response to nerve stimulation. These effects indicated that colitis induced a down-regulation of inhibitory beta3 adrenergic control of colonic smooth muscle function. This loss of adrenergic regulation may contribute to the diarrhea in inflammatory bowel disease.

The role of IL-4 in Heligmosomoides polygyrus-induced alterations in murine intestinal epithelial cell function.

IL-4 and IL-13 promote gastrointestinal worm expulsion, at least in part, through effects on nonlymphoid cells, such as intestinal epithelial cells. The role of IL-4/IL-13 in the regulation of intestinal epithelial function during Heligmosomoides polygyrus (Hp) infection was investigated in BALB/c mice infected with Hp or treated with a long-lasting formulation of recombinant mouse IL-4/alphaIL-4 complexes (IL-4C) for 7 days. Separate groups of BALB/c mice were drug-cured of initial infection and later reinfected and treated with anti-IL-4R mAb, an antagonist of IL-4 and IL-13 receptor binding, or with a control mAb. Segments of jejunum were mounted in Ussing chambers, and short circuit current responses to acetylcholine, histamine, serotonin, PGE2, and glucose were determined. Although only modest changes in epithelial cell function were observed during primary Hp infection, IL-4C or a secondary Hp infection each induced more dramatic changes, including increased mucosal permeability, reduced sodium-linked glucose absorption, and increased Cl- secretory response to PGE2. Some, but not all, effects of IL-4C and Hp infection were dependent on enteric nerves. Hp-induced changes in epithelial function were attenuated or prevented by anti-IL-4R mAb. Thus, IL-4/IL-13 mediate many of the effects of Hp infection on intestinal epithelial cell function and do so both through direct effects on epithelial cells and through indirect, enteric nerve-mediated prosecretory effects. These immune system-independent effector functions of IL-4/IL-13 may be important for host protection against gastrointestinal nematodes.

Pro- and anti-inflammatory gene expression in the murine small intestine and liver after chronic exposure to alcohol.

BACKGROUND: Endotoxin has been proposed to play a primary role in ALD, by initiating an inflammatory cascade within the liver. Although the source of these cytokines has been presumed to be circulating monocytes or tissue macrophages, ethanol-induced, nonhepatic sources of soluble mediators recently have been identified. One potential, but not clearly defined, extrahepatic source of cytokines in ALD is the intestine. In the current study, we hypothesized that alcohol would alter cytokine expression within the small intestine of mice exposed to ethanol and that LPS would alter levels of cytokine expression even more dramatically. METHODS: Mice were fed a modified Lieber-DeCarli liquid ethanol or control diet for up to 14 days prior to injecting either saline or LPS. Plasma alanine aminotransferase (ALT) and cytokine levels, histology, and RT-PCR of pro- and anti-inflammatory cytokine gene expression were determined from distal ileum and liver samples. Translocation of intestinal bacterial flora also was assessed. RESULTS: Ethanol exposure upregulated basal gene expression of IL-1 beta, TNF-alpha, IL-6, and iNOS in the distal ileum, but similar effects of ethanol on the liver were not observed. In contrast, LPS challenge of ethanol-exposed mice increased intestinal gene expression of some cytokines, but decreased expression of others. These effects were not associated with bacterial translocation. Also, ethanol alone induced a modest increase in both ICAM-1 and TLR4 mRNA expression in the intestine, but expression of both molecules was inhibited in mice that received both ethanol and LPS. Finally, whereas basal levels of hepatic IL-11 mRNA were not elevated by exposure to ethanol, intestinal IL-11 mRNA levels were increased more than 100-fold. CONCLUSIONS: These studies are the first to show that ethanol affects cytokine gene expression in the ileum and identifies the ileum as a potential target for ethanol effects. In addition, our results suggest that IL-11 expression may be enhanced in the intestine to help repair or protect this organ from alcohol-induced damage. Collectively, these studies suggest that both pro- and anti-inflammatory soluble mediators in the intestine maintain and exacerbate the local hepatic response to ethanol.

Alterations in spontaneous contractions in vitro after repeated inflammation of rat distal colon.

In inflammatory bowel disease, smooth muscle function reportedly varies with disease duration. The aim of these studies was to determine changes in the control of spontaneous contractions in a model of experimental colitis that included reinflammation of the healed area. The amplitude and frequency of spontaneous contractions in circular smooth muscle were determined after intrarectal administration of trinitrobenzenesulfonic acid in rat distal colon. With the use of a novel paradigm, rats were studied 4 h (acute) or 28 days (healed) after the initial inflammation. At 28 days, rats were studied 4 h after a second inflammation (reinflamed) of the colon. Colitis induced transient increases in the amplitude of spontaneous contractions coincident with a loss of nitric oxide synthase activity. The frequency of contractions was controlled by constitutive nitric oxide in controls. Frequency was increased in healed and reinflamed colon and was associated with a shift in the dominance of neural constitutive nitric oxide synthase control to that of inducible nitric oxide synthase (iNOS). The initial colitis induced a remodeling of the neural control of spontaneous contractions reflecting changes in their regulation by constitutive nitric oxide synthase and iNOS.

Sustained nitric oxide production via l-arginine administration ameliorates effects of intestinal ischemia-reperfusion.

BACKGROUND: The role of nitric oxide in intestinal ischemia-reperfusion is unclear-some studies link it to the harmful effects of ischemia-reperfusion, while others report it to be protective. We propose that nitric oxide levels diminish in the reperfusion period in conjunction with the onset of increased capillary permeability. The aim of this study is to determine the effect of supplementing nitric oxide synthase with its substrate, l-arginine, on development of local mucosal injury and systemic capillary leak. MATERIALS AND METHODS: Rats underwent 30 min of superior mesenteric artery occlusion followed by 4 h of reperfusion. The vehicle groups received l-arginine either intravenously (4 mg/kg/min) or into the intestinal lumen. The intravenous groups received l-arginine either before the ischemic event or after 30 min of reperfusion. Capillary leak in the gut and lung were measured, as were degree of mucosal injury and number of infiltrating neutrophils. Appropriate controls were performed. RESULTS: Thirty minutes of mesenteric ischemia followed by 4 h of reperfusion significantly increased gut and lung leak, neutrophil infiltration, and the severity of mucosal injury. l-Arginine given iv prior to ischemia inhibited lung leak, mucosal injury, and neutrophil infiltration. When arginine was given during the reperfusion period, lung leak and neutrophil infiltration but not mucosal injury were reduced. Intraluminal l-arginine reduced mucosa injury, but had no effect on capillary leak. CONCLUSIONS: Supplementation with l-arginine enhances NO production, resulting in reduced systemic endothelial dysfunction. This may act as a useful clinical adjunct in the management of trauma patients in preventing the development of ARDS and multiple organ failure.

Progressive alterations in circular smooth muscle contractility in TNBS-induced colitis in rats.

The present study was designed to investigate inflammation-induced changes in smooth muscle responses to acetylcholine and the tachykinins that may contribute to the abnormal motility associated with inflammatory bowel disease. Colitis was induced in male Sprague-Dawley rats by intrarectal administration of trinitrobenzenesulphonic acid in ethanol. After either 4 h (acute) or 7 days (chronic) the distal colon was taken for in vitro measurement of smooth muscle tension and histological assessment. Acute colitis featured injury and neutrophilic infiltration confined to the mucosa while chronic inflammation showed marked injury, lymphocytic infiltration and muscle thickening. Acute inflammation increased responses to substance P and acetylcholine but decreased responses to neurokinin A. The enhanced response to substance P was dependent on nerves, while the decreased response to neurokinin A reflected a reduction in activity at the level of the smooth muscle. In the saline group, there was evidence of cholinergic interaction with substance P, but not neurokinin A. Substance P modulation of cholinergic nerves was absent in acute inflammation. Responses to all neurotransmitters were decreased in the chronic stage. These data demonstrate progressive changes in the smooth muscle function during acute and chronic colitis that may contribute to the abnormal motility associated with inflammatory bowel disease.

Anti-inflammatory effects of U74389F in a rat model of intestinal ischemia/reperfusion injury.

OBJECTIVE: To investigate the role of eicosanoid generation and neutrophilic infiltration in the protective effects of U74389F against ischemia/reperfusion injury in the small intestines of rats. DESIGN: Prospective, randomized, controlled study. SETTING: University research laboratory. SUBJECTS: Adult, male Sprague-Dawley rats weighing between 200 and 300 g. INTERVENTIONS: Groups (5-8) of rats treated with U74389F or vehicle were subjected to a sham operation and 30 mins of ischemia by occlusion of the superior mesenteric artery or 30 mins of ischemia followed by 60 or 120 mins of reperfusion. U74389F (2.5 mg/kg i.v.) or vehicle (citrate buffer) were slowly injected 2 mins before ischemia. MEASUREMENTS AND MAIN RESULTS: Ischemia significantly (p < .05) increased mucosal injury (0 [normal] to 5) in both U74389F and untreated rats. In contrast, U74389F significantly (p < .05) attenuated the severity of injury after reperfusion. In vehicle-treated rats, ischemia/reperfusion significantly reduced villus height in both U74389F and untreated groups. However, the surface epithelial layer was intact in the U74389F but not in the vehicle-treated group. In addition, compared with the vehicle-treated group, U74389F significantly reduced neutrophil infiltration and prevented the increase in leukotriene B4 and prostaglandin E2 in response to ischemia and reperfusion. CONCLUSIONS: This study demonstrates that the mechanism of U74389F against mesenteric ischemia/reperfusion includes a delay and reduction of neutrophilic infiltrate, an inhibition of leukotriene B4 production, and a facilitation of mucosal restitution.

Neutrophil priming state predicts capillary leak after gut ischemia in rats.

BACKGROUND: Multiple organ failure after serious injury or illness is a major determinant of mortality. An initial insult is believed to "prime" circulating neutrophils and induce systemic inflammation. Thereafter, a second insult will precipitate distant organ injury. The aim of these studies was to evaluate circulating neutrophil function after mesenteric ischemia-reperfusion to determine the neutrophil "priming state," a quantitative and clinically useful predictor of multiple organ failure. MATERIALS AND METHODS: Anesthetized Sprague-Dawley rats underwent superior mesenteric artery occlusion for 30 min or sham operation and were euthanized after 2, 6, or 24 h of reperfusion. Control animals had blood taken without any intervention. To determine changes in lung capillary permeability, another group of rats received Evan's blue, a dye that binds albumin, 1 h before sacrifice. Flow cytometric analysis was performed on 5 million white blood cells after removal of red cells by lysis and centrifugation. Neutrophil number, oxidative burst, and CD18 expression were measured. RESULTS: The number of circulating neutrophils was elevated similarly in rats subjected to sham operation or ischemia-reperfusion. Oxidative burst potential was increased at 2 h, maximum at 6 h, and normal at 24 h after reperfusion, but not in sham rats. CD18 expression was similar in all groups. There was a significant temporal correlation between the "priming state" of the circulating neutrophil, defined as the product of the neutrophil number times oxidative burst, and lung leak. CONCLUSIONS: The neutrophil "priming state" may allow the clinician to better predict those patients at greatest risk for multiple organ failure.

Hyperthermia prevents functional, histological and biochemical abnormalities induced during ileitis.

Inflammatory bowel disease is associated with altered intestinal motility and epithelial damage. Hyperthermia induces heat shock protein expression, components of a basic cellular defence mechanism, and consequently prevents ischaemic damage. Here we investigate whether hyperthermia may prevent altered smooth muscle function as well as underlying inflammation in a model of inflammatory bowel disease. Ileal heat shock protein expression was induced in rats by hyperthermic shock (41.5 degrees C; 5 min). Two hours after heating or sham treatment, ileitis was evoked by TNBS. Ileal samples were taken 4 h later to determine the contractile response of circular muscle strips, and to measure heat shock protein expression, LTB4 generation and damage/inflammation. Ileitis was associated with an increase in the contractile response of circular muscle to substance P but not neurokinin A or nerve stimulation. Hyperthermia induced heat shock protein expression and also prevented this functional change as well as TNBS-induced LTB4 production, subsequent infiltration of neutrophils and epithelial damage. Thus, intestinal inflammation is associated with alterations in tachykinergic control of smooth muscle as well as inflammatory changes. Hyperthermia prevents these changes and induces heat shock protein expression. Pharmacological induction of these proteins may offer a novel clinical strategy in treating both of these aspects of disease.