Non-apoptotic TRAIL function modulates NK cell activity during viral infection.

The role of death receptor signaling for pathogen control and infection-associated pathogenesis is multifaceted and controversial. Here, we show that during viral infection, tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) modulates NK cell activity independently of its pro-apoptotic function. In mice infected with lymphocytic choriomeningitis virus (LCMV), Trail deficiency led to improved specific CD8+ T-cell responses, resulting in faster pathogen clearance and reduced liver pathology. Depletion experiments indicated that this effect was mediated by NK cells. Mechanistically, TRAIL expressed by immune cells positively and dose-dependently modulates IL-15 signaling-induced granzyme B production in NK cells, leading to enhanced NK cell-mediated T cell killing. TRAIL also regulates the signaling downstream of IL-15 receptor in human NK cells. In addition, TRAIL restricts NK1.1-triggered IFNγ production by NK cells. Our study reveals a hitherto unappreciated immunoregulatory role of TRAIL signaling on NK cells for the granzyme B-dependent elimination of antiviral T cells.

The proapoptotic Bcl-2 family member Bim plays a central role during the development of virus-induced hepatitis.

The proapoptotic Bcl-2 homolog Bim was shown to control the apoptosis of both T cells and hepatocytes. This dual role of Bim might be particularly relevant for the development of viral hepatitis, in which both the sensitivity of hepatocytes to apoptosis stimuli and the persistence of cytotoxic T cells are essential factors for the outcome of the disease. The relevance of Bim in regulating survival of cytotoxic T cells or induction of hepatocyte death has only been investigated in separate systems, and their relative contributions to the pathogenesis of T cell-mediated hepatitis remain unclear. Using the highly dynamic model system of lymphocytic choriomeningitis virus-mediated hepatitis and bone marrow chimeras, we found that Bim has a dual role in the development of lymphocytic choriomeningitis virus-induced, T cell-mediated hepatitis. Although the absence of Bim in parenchymal cells led to markedly attenuated liver damage, loss of Bim in the lymphoid compartment moderately enhanced hepatitis. However, when both effects were combined in Bim(-/-) mice, the effect of Bim deficiency in the lymphoid compartment was overcompensated for by the reduced sensitivity of Bim(-/-) hepatocytes to T cell-induced apoptosis, resulting in the protection of Bim(-/-) mice from hepatitis.

Why the structure but not the activity of the immunoproteasome subunit low molecular mass polypeptide 2 rescues antigen presentation.

The proteasome is responsible for the generation of most epitopes presented on MHC class I molecules. Treatment of cells with IFN-γ leads to the replacement of the constitutive catalytic subunits ß1, ß2, and ß5 by the inducible subunits low molecular mass polypeptide (LMP) 2 (ß1i), multicatalytic endopeptidase complex-like-1 (ß2i), and LMP7 (ß5i), respectively. The incorporation of these subunits is required for the production of numerous MHC class I-restricted T cell epitopes. The structural features rather than the proteolytic activity of an immunoproteasome subunit are needed for the generation of some epitopes, but the underlying mechanisms have remained elusive. Experiments with LMP2-deficient splenocytes revealed that the generation of the male HY-derived CTL-epitope UTY(246-254) was dependent on LMP2. Treatment of male splenocytes with an LMP2-selective inhibitor did not reduce UTY(246-254) presentation, whereas silencing of ß1 activity increased presentation of UTY(246-254). In vitro degradation experiments showed that the caspase-like activity of ß1 was responsible for the destruction of this CTL epitope, whereas it was preserved when LMP2 replaced ß1. Moreover, inhibition of the ß5 subunit rescued the presentation of the influenza matrix 58-66 epitope, thus suggesting that a similar mechanism can apply to the exchange of ß5 by LMP7. Taken together, our data provide a rationale why the structural property of an immunoproteasome subunit rather than its activity is required for the generation of a CTL epitope.

The proteasome inhibitor bortezomib enhances the susceptibility to viral infection.

The proteasome, a multicatalytic protease, is responsible for the generation of most MHC class I ligands. Bortezomib, a proteasome inhibitor, is clinically approved for treatment of multiple myeloma and mantle cell myeloma. In the present study, we investigated the effect of bortezomib on viral infection. Infection of bortezomib-treated mice with the lymphocytic choriomeningitis virus (LCMV) led to a decreased cytotoxic T cell response to several LCMV-derived CD8(+) T cell epitopes. Bortezomib treatment caused a reduced expansion of CD8(+) T lymphocytes and increased viral titers in LCMV-infected mice. Administration of bortezomib during expansion of CD8(+) T cells had no influence on the cytotoxic T cell response, suggesting that bortezomib interferes with priming of naive T cells. Indeed, determination of Ag load in spleen 4 days post infection, revealed a reduced presentation of LCMV-derived cytotoxic T cell epitopes on MHC class I molecules. In summary, we show that proteasome inhibition with bortezomib led to an increased susceptibility to viral infection, and demonstrate for the first time, that proteasome inhibitors can alter Ag processing in vivo.

From amino acid mixtures to peptides in liquid sulphur dioxide on early Earth.

The formation of peptide bonds is one of the most important biochemical reaction steps. Without the development of structurally and catalytically active polymers, there would be no life on our planet. However, the formation of large, complex oligomer systems is prevented by the high thermodynamic barrier of peptide condensation in aqueous solution. Liquid sulphur dioxide proves to be a superior alternative for copper-catalyzed peptide condensations. Compared to water, amino acids are activated in sulphur dioxide, leading to the incorporation of all 20 proteinogenic amino acids into proteins. Strikingly, even extremely low initial reactant concentrations of only 50 mM are sufficient for extensive peptide formation, yielding up to 2.9% of dialanine in 7 days. The reactions carried out at room temperature and the successful use of the Hadean mineral covellite (CuS) as a catalyst, suggest a volcanic environment for the formation of the peptide world on early Earth.

Acute cortisol administration increases sleep depth and growth hormone release in patients with major depression.

Acute administration of cortisol increases non-rapid-eye movement (non-REM) sleep, suppresses rapid-eye movement (REM) sleep and stimulates growth hormone (GH) release in healthy subjects. This study investigates whether cortisol has similar endocrine and electrophysiological effects in patients with depression who typically show a pathological overactivity of the hypothalamus-pituitary-adrenal (HPA) system. Fifteen depressed inpatients underwent the combined dexamethasone/corticotropin-releasing hormone test followed by three consecutive sleep EEG recordings in which the patients received placebo (saline) and hourly injections of cortisol (1mg/KG BW). Cortisol increased duration and intensity of non-REM sleep in particular in male patients and stimulated GH release. The activity of the HPA axis appeared to influence the cortisol-induced effects on non-REM sleep and GH levels. Stimulation of delta sleep was less pronounced in patients with dexamethasone nonsuppression. In contrast, REM sleep parameters were not affected by the treatment. These data demonstrate that the non-REM sleep-promoting effects of acute cortisol injections observed in healthy controls could be replicated in patients with depression. Our results suggest that non-REM and REM sleep abnormalities during the acute state of the disease are differentially linked to the activity of the HPA axis.

Dex/CRH-test response and sleep in depressed patients and healthy controls with and without vulnerability for affective disorders.

Sleep electroencephalographic (EEG) abnormalities and increased hypothalamo-pituitary-adrenal (HPA) axis activity are the most prominent neurobiological findings in depression and were suggested as potential biomarker for depression. In particular, increased rapid eye movement sleep (REM) density, deficit in slow wave sleep and excessive stress hormone response are associated with an unfavorable long-term outcome of depression. Recent studies indicate that the sleep and endocrine parameters are related to each other. This study investigated the association of sleep structure including a quantitative EEG analysis with the results of the combined dexamethasone (Dex)/corticotropin-releasing hormone (CRH)-test in 14 patients with a severe major depression, 21 healthy probands with a positive family history of depression (HRPs) and 12 healthy control subjects without personal and family history for psychiatric disorders. As expected patients with depression showed an overactivity of the HPA axis, disturbed sleep continuity and prolonged latency until slow wave sleep in the first sleep cycle. Differences in microarchitecture of sleep were less prominent and restricted to a higher NonREM sigma power in the HRP group. Dexamethasone suppressed cortisol levels were positively associated with higher NonREM sigma power after merging the three groups. We also observed an inverse association between the ACTH response to the Dex/CRH-test and rapid eye movement sleep (REM) density in HRPs, with suggestive evidence also in patients, but not in controls. This contra-intuitive finding might be a result of the subject selection (unaffected HRPs, severely depressed patients) and the complementarity of the two markers. HRPs and patients with high disease vulnerability, indicated by an elevated REM density, seem to have a lower threshold until an actual disease process affecting the HPA axis translates into depression, and vice versa. To summarize, our findings provide further evidence that the HPA axis is involved in the sleep regulation in depression. These associations, however, are not unidimensional, but dependent on the kind of sleep parameters as well as on the selection of the subjects.

Strength-size relationships in two porous biological materials.

According to the Weibull theory for brittle materials, the mean experimental strength decreases with test specimen size. For the brittle parts of an organism this would mean that becoming larger in size results automatically in reducing strength. This unfavorable relationship was investigated for two porous, biological materials that are promising concept generators for crack deflective and energy dissipative applications in compressive overloading: the quasi-brittle coconut endocarp and the brittle spines of the sea urchin Heterocentrotus mamillatus. Segments in different volumes were prepared and tested in uniaxial compression experiments. Failure of both materials is Weibull distributed underlining that it is caused by statistically distributed flaws in the structure. However, the coconut endocarp has a much higher Weibull modulus (m = 14.1-16.5) than the spines (m = 5). The more predictable failure of the endocarp is probably attributed to a rather homogeneous microstructural design and water bound in the structure. In terms of the spines it was found that the Weibull modulus is structure dependent: More homogeneous spines feature a higher Weibull modulus than spines with a heterogeneous structure. Whereas the nearly dense endocarp exhibited, although less pronounced, the expected decrease in strength with increase in size, the spines showed a failure independently of size. This remarkable behavior may be explained with their highly porous internal structure. Small and large spines consist of struts of similar size, which constitute the porous internal structure, potentially limiting the flaw size to the size of the strut regardless of the spine size. STATEMENT OF SIGNIFICANCE: Scaling is an important aspect of the biomimetic work process, since biological role models and structures have rarely the same size as their technical implementations. The algorithms of Weibull are a standard tool in material sciences to describe scaling effects in materials whose critical strength depends on statistically distributed flaws. The challenge is to apply this theory (developed for homogeneous, isotropic technical materials) to brittle and quasi-brittle biological materials with hierarchical structuring. This study is a first approach to verify whether the Weibull theory can be applied to the coconut endocarp and to sea urchin spines in order to model their size/volume/property-relations.

Strength, elasticity and the limits of energy dissipation in two related sea urchin spines with biomimetic potential.

The calcitic spines of the sea urchins Heterocentrotus mamillatus and H. trigonarius are promising role models for lightweight applications, bone tissue scaffolds and energy dissipating processes due to their highly porous and organized structure. Therefore, mechanical properties including Young's Modulus, strength, failure behaviour and energy dissipation efficiency have been investigated in depth with uniaxial compression experiments, 3-point bending tests and resonance frequency damping analysis. It was found that despite a very similar structure, H. trigonarius has a significantly lower porosity than H. mamillatus leading to a higher strength and Young's Moduli, but limited ability to dissipate energy. In order to show reliable energy dissipation during failure in uniaxial compression, a transition porosity of 0.55-0.6 needs to be exceeded. The most effective structure for this purpose is a homogeneous, foam-like structure confined by a thin and dense shell that increases initial strength and was found in numerous spines of H. mamillatus. Sharp porosity changes induced by dense growth layers or prominent wedges of the spines' radiating building principle act as structural weaknesses, along which large flakes can be spalled, reducing the energy dissipation efficiency considerably. The high strength and Young's Modulus at the biologically necessary high porosity levels of the spines is useful for Heterocentrotus and their construction therefore remains to be a good example of biomimetics. However, the energy dissipative failure behaviour may be regarded as a mere side effect of the structure.

Associations of nocturnal sleep with experimental pain and pain catastrophizing in healthy volunteers.

Strong alterations of night sleep (e.g., sleep deprivation, insomnia) have appeared to affect pain in inducing hyperalgesic changes. However, it has remained unclear whether everyday variations of night sleep in healthy individuals have any influence on pain processing. Forty healthy subjects were studied by portable polysomnography (PSG) and sleep questionnaire during two non-consecutive nights at home. Experimental pain parameters (pressure pain threshold, temporal summation = TS, conditioned pain modulation = CPM) and situational pain catastrophizing (Situational Catastrophizing Questionnaire = SCQ) were always assessed the evening before and the morning after sleep recording in a pain laboratory. Linear regression analyses were computed to test the prediction of overnight changes in pain by different sleep parameters. Significant prediction of changes in pain parameters by sleep parameters was limited (2 out of 12 analyses), indicating that everyday variations in sleep under non-pathological and low stress conditions are only weakly associated with pain.

Immune-endocrine host response to endotoxin in major depression.

OBJECTIVE: An impaired hypothalamic-pituitary-adrenocortical (HPA) function is a well-established finding in major depression (MD), but it is still unclear how this dysfunction affects immune responses in this disorder. METHOD: To further examine the relationship between immune and endocrine responses in MD, 0.4ng/kg body weight endotoxin [LPS] or 100mug hCRH were sequentially applied to 12 patients with MD and to 12 age- and gender-matched healthy controls after pre-treatment with 1.5mg dexamethasone (DEX). Immune (TNF-alpha, IL-6, rectal temperature) and endocrine (ACTH, cortisol) parameters were examined as area under the curve (AUC) levels. RESULTS: After pre-treatment with DEX, LPS evoked an immune response in all participants of the study with most immune parameters significantly related to the endotoxin challenge. However, only a marked immune response resulted in an additional endocrine reaction. Subsequently, the quantitative extent of the endocrine reaction was related to the extent of the immune response after DEX/LPS challenge. Pre-LPS AUC levels of cortisol, ACTH and post-LPS levels of IL-6 as well as the post-CRH AUC levels of cortisol and ACTH were related to the depressive symptomatology as measured by the Beck depression inventory (BDI). In depressive patients who showed increased cortisol plasma levels before LPS, the later increase in IL-6 was reduced. CONCLUSIONS: The challenge with DEX/LPS did not reveal major impairments of evoked immune functions in MD. Only the endocrine parameters and the IL-6 response were related to the depressive symptomatology, suggesting a limited interaction between immune and endocrine dysfunctions in MD.

No Impaired Glucose Tolerance in Primary Insomnia Patients with Normal Results of Polysomnography.

BACKGROUND: According to recent studies, sleep restriction and disruption both have a prominent negative influence on glucose metabolism. This could also be shown in sleep disorders, such as sleep apnea and the restless legs syndrome. However, similar studies regarding insomnia have not been that consistent, yet. Moreover, most previous studies did not include objective polysomnography (PSG) data. METHODS: Patients with primary insomnia (N = 17) and healthy controls (N = 15) were investigated using psychometric tests such as the Epworth Sleepiness Scale and the Pittsburgh sleep quality index (PSQI). Two nights of full PSG were performed in all subjects, and after the first PSG night subjects underwent a standard oral glucose tolerance test (OGTT). PSG-, arousal-, and glucose metabolism-parameters were compared between groups. RESULTS: Patients with insomnia were, as expected, sleepier than healthy controls and showed higher PSQI values. All PSG parameters, however, including parameters related to nocturnal arousals, did not differ between groups. Moreover, OGGT results and all other parameters of glucose tolerance were not different between insomniac patients and healthy controls. CONCLUSION: Our findings suggest that glucose tolerance is not impaired in patients with chronic insomnia and normal PSG-findings. Therefore, impaired glucose metabolism and diabetes related to insomnia in earlier studies might be restricted to those patients who have objectively disturbed sleep.

The Munich vulnerability study on affective disorders: premorbid polysomnographic profile of affected high-risk probands.

BACKGROUND: The most characteristic alterations in the sleep electroencephalogram (EEG) during major depression are a shortened latency to rapid eye movement (REM) sleep and an elevated REM density. Because these changes persist in remission, they might represent vulnerability markers. To identify vulnerability markers, we investigated premorbid sleep EEG parameters in healthy high-risk probands (HRPs) with a positive family history of affective disorders. METHODS: We identified 136 depressed inpatients from referrals to our hospital who had first-degree relatives with an affective disorder as well as first-degree relatives with no current or lifetime history of psychiatric disorders. The latter (the HRP group) were investigated by polysomnography. During the follow-up period, 20 HRPs developed an affective disorder. Their premorbid sleep data were analyzed. RESULTS: Premorbid sleep EEG of affected HRPs showed an increased REM density (total night and first REM period) compared with the control group without personal or family history of a psychiatric disorder. CONCLUSIONS: Increased REM density can be observed not only in patients with depression, but also in their healthy relatives. Moreover, it is predictive for the onset of a psychiatric disorder. Therefore, it can be recommended as a possible endophenotype of these diseases.

The Munich Vulnerability Study on Affective Disorders: stability of polysomnographic findings over time.

BACKGROUND: Some of the sleep abnormalities found in depression also persist in remission, suggesting that these parameters could represent trait or vulnerability markers. In a previous study, we found that about one third of a group of high-risk probands (HRPs) showed sleep patterns that were comparable to those of depressed patients. In the present study, we re-investigated a subsample of these HRPs to evaluate the stability of these findings over time. METHODS: We investigated the sleep-electroencephalograms of 82 healthy subjects with a high genetic load of affective disorders. We were able to re-investigate 26 of these HRPs after a mean interval of 3.5 years. Thirty-five unrelated control probands and 33 unrelated depressed inpatients that were recruited at the first investigation served as reference groups. RESULTS: At index investigation, we found that the HRPs showed a significantly increased rapid eye movement (REM) sleep density compared to control subjects. At the second examination, no changes of the polysomnographic observations over time could be observed; in particular, the REM density remained elevated. CONCLUSIONS: The increased REM density in high-risk subjects for an affective disorder at index investigation was stable over time, so that one of the requirements for a true vulnerability marker is fulfilled.

Sleep in eating disorders.

Sleep research on eating disorders has addressed two major questions: (1) the effects of chronic starvation in anorexia nervosa and of rapidly fluctuating eating patterns in bulimia nervosa on the sleep regulating processes and (2) the search for a significant neurobiological relationship between eating disorders and major depression. At present, the latter question appears to be resolved, since most of the available evidences clearly underline the notion that eating disorders (such as anorexia and bulimia nervosa) and affective disorders are two distinct entities. Regarding the effects of starvation on sleep regulation, recent research in healthy humans and in animals demonstrates that such a condition results in a fragmentation of sleep and a reduction of slow wave sleep. Although several peptides are supposed to be involved in these regulatory processes (i.e. CCK, orexin, leptin), their mode of action is still poorly understood. In opposite to these experimentally induced sleep disturbances are the findings that the sleep patterns in eating disorder patients per se do not markedly differ from those in healthy subjects. However, when focusing on the so-called restricting anorexics, who maintain their chronic underweight by strictly dieting, the expected effects of malnutrition on sleep can be ascertained. Furthermore, at least partial weight restoration results in a 'deepening' of nocturnal sleep in the anorexic patients. However, our knowledge about the neurobiological systems (as well as their circadian pattern of activity) that transmit the effects of starvation and of weight restoration on sleep is still limited and should be extended to metabolic signals mediating sleep.

The Munich Vulnerability Study on Affective Disorders: risk factors for unipolarity versus bipolarity.

BACKGROUND: An individual with a high genetic load for psychiatric disorders is subject to a considerable risk factor for an affective illness. Family studies usually try to distinguish between bipolar and unipolar disorders since it was suggested that they might show different modes of inheritance. The aim of this study was to differentiate between healthy members of unipolar and bipolar families without a previous history of any psychiatric disorder according to the neurobiological and psychometric findings. METHODS: We first analysed the results obtained from neurobiological and psychometric measurements taken from 75 healthy subjects who had at least two close relatives with a unipolar and a bipolar disorder. In a second step we examined the subjects with a parental affective disorder; finally, we compared the members of 'pure' unipolar, bipolar and of mixed families to each other. RESULTS: The first-degree relatives of unipolar patients showed a significantly higher REM density and scored higher on scales of 'neuroticism' and 'vegetative lability' than the controls. No significant differences could be noticed between the relatives of unipolar and bipolar patients, either when considering the degree of relationship, or the parental type of affective disorder and the 'purity' of the respective families. CONCLUSIONS: We found some distinct neurobiological and psychometric differences between the relatives of unipolar patients and the control probands. No obvious differences, however, were ascertained between relatives of unipolar and bipolar patients. Therefore, we consider it to be possible that these findings represent potential vulnerability markers for affective disorders in general.

Impaired glucose tolerance in sleep disorders.

BACKGROUND: Recent epidemiological and experimental data suggest a negative influence of shortened or disturbed night sleep on glucose tolerance. Due to the high prevalence of sleep disorders this might be a major health issue. However, no comparative studies of carbohydrate metabolism have been conducted in clinical sleep disorders. METHODOLOGY/PRINCIPAL FINDINGS: We performed oral glucose tolerance tests (OGTT) and assessed additional parameters of carbohydrate metabolism in patients suffering from obstructive sleep apnea syndrome (OSAS, N = 25), restless legs syndrome (RLS, N = 18) or primary insomnia (N = 21), and in healthy controls (N = 33). Compared to controls, increased rates of impaired glucose tolerance were found in OSAS (OR: 4.9) and RLS (OR: 4.7) patients, but not in primary insomnia patients (OR: 1.6). In addition, HbA1c values were significantly increased in the same two patient groups. Significant positive correlations were found between 2-h plasma glucose values measured during the OGTT and the apnea-arousal-index in OSAS (r = 0.56; p<0.05) and the periodic leg movement-arousal-index in RLS (r = 0.56, p<0.05), respectively. Sleep duration and other quantitative aspects of sleep were similar between patient groups. CONCLUSIONS/SIGNIFICANCE: Our findings suggest that some, but not all sleep disorders considerably compromise glucose metabolism. Repeated arousals during sleep might be a pivotal causative factor deserving further experimental investigations to reveal potential novel targets for the prevention of metabolic diseases.

Immune cell-mediated liver injury.

Liver diseases represent an important cause of morbidity and mortality in the world. Death of hepatocytes and other hepatic cell types is a characteristic feature of several forms of liver injury such as cholestasis, viral hepatitis, drug- or toxin-induced injury, and alcohol-induced liver damage. Moreover, irrespectively of the reason, liver injury seems to be facilitated by similar immune effector mechanisms common to these various liver diseases. Indeed, common immune effector mechanisms may explain the high prevalence of cirrhosis and cancer development in most forms of liver disease. Improved understanding of the immune cell-mediated mechanisms involved in hepatocyte cell death could be beneficial for the development of common therapeutic strategies against different forms of liver diseases. In this review, we will discuss novel findings on the role of different immune cells in liver disease and immune cell-induced death executioner mechanisms involved in hepatocyte cell death.

A selective inhibitor of the immunoproteasome subunit LMP7 blocks cytokine production and attenuates progression of experimental arthritis.

The immunoproteasome, a distinct class of proteasome found predominantly in monocytes and lymphocytes, is known to shape the antigenic repertoire presented on class I major histocompatibility complexes (MHC-I). However, a specific role for the immunoproteasome in regulating other facets of immune responses has not been established. We describe here the characterization of PR-957, a selective inhibitor of low-molecular mass polypeptide-7 (LMP7, encoded by Psmb8), the chymotrypsin-like subunit of the immunoproteasome. PR-957 blocked presentation of LMP7-specific, MHC-I-restricted antigens in vitro and in vivo. Selective inhibition of LMP7 by PR-957 blocked production of interleukin-23 (IL-23) by activated monocytes and interferon-gamma and IL-2 by T cells. In mouse models of rheumatoid arthritis, PR-957 treatment reversed signs of disease and resulted in reductions in cellular infiltration, cytokine production and autoantibody levels. These studies reveal a unique role for LMP7 in controlling pathogenic immune responses and provide a therapeutic rationale for targeting LMP7 in autoimmune disorders.