Balance between IL-3 and type Iinterferons and their interrelationship with FasL dictates lifespan and effector functions of human basophils.

BACKGROUND: In contrast to eosinophils and neutrophils, the regulation of the lifespan of human basophils is poorly defined, with the exception of the potent anti-apoptotic effect of IL-3 that also promotes pro-inflammatory effector functions and phenotypic changes. Type I IFNs (IFN-α, IFN-ß), which are well known for their anti-viral activities, have the capacity to inhibit allergic inflammation. OBJECTIVE: To elucidate whether type I IFNs have the potential to abrogate the lifespan and/or effector functions of human basophils. METHODS: We cultured human basophils, and for comparison, eosinophils and neutrophils, with IL-3, interferons, FasL and TRAIL, alone or in combination, and studied cell survival, effector functions and signalling pathways involved. RESULTS: Despite an identical pattern of early signalling in basophils, eosinophils and neutrophils in response to different types of interferons, only basophils displayed enhanced apoptosis after type I IFN treatment. IFN-γ prolonged survival of eosinophils but did not affect the lifespan of basophils. IFN-α-mediated apoptosis required STAT1-STAT2 heterodimers and the contribution of constitutive p38 MAPK activity. Whereas the death ligands FasL and TRAIL-induced apoptosis in basophils per se, IFN-α-mediated apoptosis did neither involve autocrine TRAIL signalling nor did it sensitize basophils to FasL-induced apoptosis. However, IFN-α and FasL displayed an additive effect in killing basophils. Interestingly, IL-3, which protected basophils from IFN-α-, TRAIL- or FasL-mediated apoptosis, did not completely block the additive effect of combined IFN-α and FasL treatment. Moreover, we demonstrate that IFN-α suppressed IL-3-induced release of IL-8 and IL-13. In contrast to IFN-α-mediated apoptosis, these inhibitory effects of IFN-α were not dependent on p38 MAPK signalling. CONCLUSIONS AND CLINICAL RELEVANCE: Our study defines the unique and granulocyte-type-specific inhibitory and pro-apoptotic function of type I IFNs and their cooperation with death ligands in human blood basophils, which may be relevant for the anti-allergic properties of type I IFNs.

Gonadotrophin stimulation in IVF alters the immune cell profile in follicular fluid and the cytokine concentrations in follicular fluid and serum.

Study question: Are the immune cell profiles and the cytokine concentrations in follicular fluid (FF) and serum at the preovulatory stage different in conventional exogenous gonadotrophin stimulated IVF (c-IVF) compared with natural cycle IVF (NC-IVF)? Summary answer: The cell counts of CD45+ leucocytes and T cell subpopulations and the cytokine concentrations in FF and serum are different in c-IVF compared to NC-IVF. What is known already: FF-derived cells are heterogeneous. Immune cells are involved in intra-ovarian processes and cytokines are required for normal follicular development. Gonadotrophins stimulate the regulatory intrafollicular system and influence the local distribution of immune cells and the intrafollicular release of cytokines. Administration of exogenous gonadotrophins may have a significant effect on this local regulatory system, which then in turn could influence oocyte quality. Study design, size, duration: The study included 105 patients, 69 undergoing c-IVF and 36 undergoing NC-IVF. c-IVF was performed by exogenous ovarian stimulation with hMG and GnRH antagonists. Participants/materials, setting, methods: FF samples were collected from the first dominant follicle in c-IVF without pooling and from single leading preovulatory follicles in NC-IVF. Three different approaches were used to analyze FF samples: (i) microscopic investigation of CD45+ leucocytes, (ii) fluorescence-activated cell sorting to determine CD19+ B cells and CD3+ T cells including T cell subpopulations (CD4+, CD8+), and (iii) evaluation of tumour necrosis factor-alpha (TNF-α), interferon-gamma (INF-γ), interleukins (IL)-2, -6, -8, -10 and vascular endothelial growth factor (VEGF) levels in matched FF and serum samples using the Bio-Plex® platform. Main results and the role of chance: FF obtained from c-IVF contained proportionally more CD45+ leucocytes (P = 0.0384), but fewer CD8+ cytotoxic T cells than FF from NC-IVF. CD3+ T lymphocytes were the most common type of lymphocytes, and the number thereof was comparable in the two study groups. In c-IVF, serum VEGF levels were higher (P = 0.007) than in NC-IVF while FF contained marginally decreased concentrations of IL-8 in c-IVF in comparison to NC-IVF. The cytokine concentration gradient between FF and serum in c-IVF was 10-fold for IL-8 and 8-fold for VEGF and thereby markedly lower than in NC-IVF, where the differences were 32-fold and 30-fold, respectively. Strong positive correlations were determined between FF- IL-10 and FF- VEGF in c-IVF (r = 0.85, P < 0.0001) and in NC-IVF (r = 0.81, P < 0.0001). Large scale data: N/A. Limitations, reasons for caution: The ovulation of NC-IVF follicles was induced by the exogenous administration of hCG, which means that the environment did not fully correspond to the physiological situation. Wider implications of the findings: The differences in the immune profile and the cytokine concentrations in c-IVF and NC-IVF follicles support the hypothesis that conventional ovarian stimulation affects indirectly and heterogeneously the intrafollicular milieu, and thereby possibly affects the oocyte quality and the IVF outcome. However, further studies are needed to confirm our findings and to refine stimulation protocols in the context of optimizing the intrafollicular environment during oocyte maturation. Study funding/competing interest(s): The study was supported by a research grant from IBSA Institut Biochimique SA and MSD Merck Sharp & Dohme GmbH. The authors are clinically involved in low dose mono-follicular stimulation and IVF-therapies, using gonadotrophins from all gonadotrophins distributors on the Swiss market, including Institut Biochimique SA and MSD Merck Sharp & Dohme GmbH.

In human basophils, IL-3 selectively induces RANKL expression that is modulated by IgER-dependent and IgER-independent stimuli.

BACKGROUND: Receptor activator of NF-κB ligand (RANKL) is expressed as either surface (hRANKL1, hRANKL2) or soluble (hRANKL3) form. RANKL is involved in multifaceted processes of immunoregulation and bone resorption such as they occur in rheumatoid arthritis (RA). Interestingly, activated basophils, which are effector cells in allergic inflammation, contribute to the progress of collagen-induced arthritis (CIA), a mouse model for RA. Here, we investigate under which conditions human basophils express RANKL. METHODS: Among other stimuli, basophils were cultured with IL-3 alone. Alternatively, as a secondary stimulus, IgER-dependent or IgER-independent agents were added simultaneously either with IL-3 or after prolonged IL-3 culturing. Expression of RANKL protein and mRNA was analyzed by flow cytometry, ELISA, and real-time PCR. A coculture system was applied to investigate biological activity of basophil-derived RANKL. RESULTS: We show that in human basophils, IL-3 but no other stimulus induces de novo expression of soluble and surface RANKL, of which the latter enhances survival of MoDC. Upon simultaneous stimulation, IgER cross-linking reduces surface RANKL expression, while IgER-independent stimuli have no effect. This is in contrast to consecutive stimulation, as triggering with both IgER-dependent and IgER-independent stimuli enhances RANKL expression, particularly in its soluble form. Real-time PCR analysis shows that RANKL expression is mainly regulated at the mRNA level. CONCLUSION: This study identifies IL-3 as a potent inducer of RANKL expression in human basophils, suggesting them to interact with bone physiology and activation of immune cells. IgER-dependent and IgER-independent stimuli modulate the IL-3-mediated RANKL expression in a time- and stimulus-dependent fashion.

IL-33 is a mediator rather than a trigger of the acute allergic response in humans.

BACKGROUND: IL-33 enhances FcεRI-induced mediator release in human basophils without inducing degranulation itself. In contrast, studies in mice suggested that in the presence of high IgE levels, IL-33 triggers degranulation and anaphylaxis of similar severity as specific allergen. Consistent with this view, sera of atopic patients contain elevated levels of IL-33 after anaphylaxis. In this study, we determined whether IL-33 is potentially anaphylactogenic in humans with high IgE levels by regulating exocytosis independent of FcεRI cross-linking. Furthermore, we investigated whether IL-33 is released upon allergen provocation in vivo. METHODS: In subjects with high serum IgE levels, we measured IL-33-induced histamine/LTC4 in vitro, CD63 translocation ex vivo, and responsiveness of mast cells in vivo by skin prick test (SPT). In asthma patients, release of IL-33 and its correlation with early (tryptase)- and late-phase markers (IL-13 levels, eosinophil numbers) of the allergic response were assessed in bronchoalveolar lavage fluids (BALFs) after allergen challenge. RESULTS: IL-33 itself does not trigger basophil degranulation in vitro and ex vivo, even in subjects with high serum IgE levels, and negative SPTs demonstrate that skin mast cells do not degranulate in response to IL-33. However, in response to allergen challenge, IL-33 is rapidly released into BALFs at levels that do not correlate with other immediate- and late-phase parameters. CONCLUSION: IL-33 is unlikely an independent trigger of anaphylaxis even in subjects with high IgE levels. However, the rapid release of IL-33 upon allergen provocation in vivo supports its role as a mediator of immediate allergic responses.

Robust expression of CCR3 as a single basophil selection marker in flow cytometry.

BACKGROUND: Basophil activation tests (BAT) rely on different combinations of basophil selection and activation markers. Whereas activation markers, especially CD63, are widely validated, the most suitable and robust marker for basophil selection is still a matter of debate. AIMS: Comparison of cell surface expression of two commonly used basophil selection markers (IgE, CD123/HLA-DR) with CCR3 in an unselected group of atopic and nonatopic donors in resting and activated basophils. METHODS: EDTA blood of 94 healthy adults, about half of them atopic by history, was analyzed using two different staining strategies: anti-CD123-PE/anti-HLA-DR-PerCP/anti-lin1-FITC and anti-IgE-FITC/anti-CD3-PerCP/anti-CCR3-PE. Additionally 40 pollen-allergic patients were recruited for the assessment of CCR3 expression after basophil activation. RESULTS: In resting basophils, cell surface expression of the three basophil selection markers was most constant for CCR3. IgE gating strategy showed the highest variation and up to 80% of nonbasophils in the selected gate in certain donors. During basophil activation, a shift of the mean fluorescence intensity for CCR3 toward the lower third of the CCR3-positive population could be demonstrated, but neither were CCR3-positive cells significantly lost for further analysis nor was differentiation between CCR3-positive and CCR3-negative cell populations hampered by this shift. CONCLUSIONS: CCR3 is a stable and highly expressed basophil selection marker, independent of the atopic background or basophil activation state and allows an accurate identification of basophils without need of a second marker. The basophil markers CD123/HLA-DR and IgE showed significantly higher interindividual variability in cell surface expression and are therefore less suited as selection markers.

Efficacy and safety of secukinumab for the treatment of severe ABCA12 deficiency-related ichthyosis in a child.

Background: Patients with severe autosomal recessive congenital ichthyosis (ARCI) show a T helper 17/interleukin 17 (Th17/IL17) skewing in their skin and serum, resembling the inflammatory profile of psoriatic patients. Secukinumab, an IL-17A inhibitor, has shown clinical efficacy in patients with moderate-to-severe plaque psoriasis. Aims: To test the clinical efficacy and safety of secukinumab in a paediatric patient with ATP-binding cassette subfamily A member 12 deficiency-related severe erythrodermic ARCI. Materials & Methods: 6-months therapeutic trial. During the first 4-weeks induction period, the patient received weekly subcutaneous injections of 150 mg secukinumab (five injections in total). During the following 20-weeks maintenance period, the patient was given a subcutaneous injection of 150 mg secukinumab every 4 weeks. Result & Discussion: After the 6-months therapy period, there was a 48% reduction from the baseline Ichthyosis-Area-Severity-Index (-Erythema/-Scaling) score. The treatment was well tolerated. Moreover, cytokine analysis revealed a reduction of keratinocyte-derived proinflammatory cytokines and an abrogation of Th17-skewing during therapy. Conclusion: Further studies are needed to evaluate the effects of the use of IL-17A inhibition in ARCI patients.

Inositol treatment of obsessive-compulsive disorder.

OBJECTIVE: Earlier studies reported that inositol, a simple polyol second messenger precursor, was effective in controlled trials for patients with depression and panic. In this study its effectiveness in obsessive-compulsive disorder was investigated. METHOD: Thirteen patients with obsessive-compulsive disorder completed a double-blind, controlled crossover trial of 18 g/day of inositol or placebo for 6 weeks each. RESULTS: The subjects had significantly lower scores on the Yale-Brown Obsessive Compulsive Scale when taking inositol than when taking placebo. CONCLUSIONS: The authors conclude that inositol is effective in depression, panic, and obsessive-compulsive disorder, a spectrum of disorders responsive to selective serotonin reuptake inhibitors.

Double-blind, placebo-controlled, crossover trial of inositol treatment for panic disorder.

OBJECTIVE: Because they found in an earlier study that inositol, an important intracellular second-messenger precursor, was effective against depression in open and double-blind trials, the authors studied its effectiveness against panic disorder. METHOD: Twenty-one patients with panic disorder with or without agoraphobia completed a double-blind, placebo-controlled, 4-week, random-assignment crossover treatment trial of 12 g/day of inositol. RESULTS: The frequency and severity of panic attacks and the severity of agoraphobia declined significantly more after inositol than after placebo administration. Side effects were minimal. CONCLUSIONS: The authors conclude that inositol's efficacy, the absence of significant side effects, and the fact that inositol is a natural component of the human diet make it a potentially attractive therapeutic for panic disorder.

Response to missile attacks on civilian targets in patients with panic disorder.

BACKGROUND: The complex interaction that exists between biological and cognitive factors determines the reaction of panic-disorder patients to stressors. The current study was conducted to systematically assess the behavioral effects of a real, life-threatening event on panic-disorder patients. METHOD: Sixty-five panic-disorder patients completed structured telephone interviews during the first 4 weeks of the Persian Gulf War. Evaluation included frequency of panic attacks, anxiety levels, and function levels both during and between air raid alarms. RESULTS: The findings indicate that panic-disorder patients, despite high levels of anxiety, did not demonstrate an increased frequency of panic attacks during the Persian Gulf War. In addition, the majority of patients reported good-to-high levels of functioning during the crisis in both everyday and alarm-related functioning. Grouping of subjects according to proximity to risk or current antipanic treatment did not produce significant differences in the frequency of panic attacks or levels of anxiety. CONCLUSION: The findings suggest that vulnerability of patients with panic disorder to a "panic-stricken" response does not increase during real-life stressors. The lack of increased frequency of panic attacks observed under these circumstances provides additional support for the opinion that panic and fear are two distinct entities.

A placebo-controlled cross-over trial of adjunctive EPA in OCD.

Several clinical studies showed beneficial effects of omega-3 fatty acids in major affective disorders, including resistant depression. Some antidepressants are also effective, albeit less so, in obsessive-compulsive disorder (OCD). We therefore undertook a preliminary placebo-controlled cross-over trial of adjunctive eicosapentaenoic acid (EPA) in OCD. Eleven patients with current obsessive-compulsive disorder, who were on a stable maximally tolerated dose of SSRI with no further improvement over at least the last two months, were recruited. Subjects were randomly allocated to begin 6 weeks of placebo (2 g liquid paraffin per day) followed by 6 weeks of 2 g of EPA or EPA followed by placebo. Patients continued their prior SSRIs at the same dose. Assessments were performed with the Yale-Brown Obsessive-Compulsive Scale (YBOCS), and the Hamilton Rating Scales for depression (HAM-D) and anxiety (HAM-A). There were no effects of order of treatment. Time had a main effect of YBOCS scores; mean scores declined from 26.0 (+/-5) to 17.6 (+/-6) by week 6 on placebo and to 18.5 (+/-4) on EPA. There were no effects on HAM-D and HAM-A. No clinically relevant side effects were reported. The results of this study suggest that adjunctive EPA is ineffective against OCD.

Acute inositol does not attenuate m-CPP-induced anxiety, mydriasis and endocrine effects in panic disorder.

Many anti-panic drugs, administered chronically, can block pharmacologically-induced "panic attacks"; acutely they often exacerbate panic disorder. Theories of action need to account for this biphasic effect. Chronic inositol had previously shown efficacy against panic disorder. The authors investigated the effect of a single dose of 20 g inositol on an m-CPP challenge in a double-blind placebo-controlled crossover trial in panic-disorder patients. Seven patients had robust psychological, physiological and endocrine responses to 0.08 mg m-CPP i.v.; inositol had virtually no effect on these responses, although it had some acute effects during the evening before the challenge. A similar trial involving chronic inositol would be of interest.

A double-blind crossover comparison of clomipramine and desipramine in the treatment of panic disorder.

OBJECTIVE: To compare the efficacy of clomipramine hydrochloride (CMI), a serotonin reuptake inhibitor with the noradrenergic tricyclic antidepressant agent, and desipramine hydrochloride (DMI) for patients with panic disorder (PD). METHOD: Following a 2-week, single-blind placebo washout phase, 17 PD outpatients completed a 16-week, double-blind, crossover comparison of CMI and DMI. Key outcome measures included panic attacks frequency, the NIMH Global Scales for Anxiety, Depression and Impairment, Hamilton Anxiety Scale (Psychic and Somatic Subscales), Zung Anxiety Inventory (Raw and Index Subscales) and the Spielberger State Anxiety Scale. RESULTS: Both CMI and DMI led to significant improvement from baseline placebo state in panic attacks frequency and behavioral ratings (p<0.001). CMI led to a greater reduction in the frequency of panic attacks (p=0.028) and was superior to DMI on ratings of anxiety: NIMH Global Anxiety, Zung Anxiety Scale (Raw and Index) and the Spielberger Anxiety Scale. No difference was found between the drugs on the NIMH Global Impairment Scale and the Hamilton Somatic and Psychic Scales. CONCLUSION: Both drugs appeared to have significant therapeutic effects in patients with PD, but CMI appeared to be more effective. The effectiveness of the serotonergic drug suggests that the role of the serotonergic system in the pathogenesis of PD should be further explored.

Realism of confidence in obsessive-compulsive checkers.

The study examined whether obsessive-compulsive (OC) checkers have reduced confidence in their knowledge. OC checkers were compared with panic disorder (PD) patients and nonpatient controls using a calibration-of-knowledge procedure. Participants completed a general knowledge questionnaire, rated their confidence in each answer, and estimated the total number of correct answers. These responses were converted to 2 measures of confidence relative to performance--over/underconfidence and over/underestimation. OC checkers had lower scores than nonpatients did on both measures, whereas the PD patients did not differ from either group. For the OC checkers, relative confidence was inversely related to the severity of obsessions. The authors speculate that confidence may depend on a confirmation bias in testing hypotheses and that the reduced confidence in OC checkers may reflect a disconfirmation bias in this population.

Pilot project for registering acute myocardial infarctions in Belgium.

In order to prepare Belgian participation in the International Study "Monitoring of trends and determinants in cardiovascular disease (MONICA)", a pilot project for registration of acute myocardial infarctions was organized in the Flemish city of Ghent and the Walloon city of Charleroi. Thanks to the collaboration of 449 general practitioners and 17 hospitals, 790 acute coronary events (fatal and non-fatal) were registrated during one year among the 251 000, 25 to 69 year old, inhabitants of Ghent and Charleroi. The registration procedures, strictly standardized between the two cities, allowed comparison of the incidence data. In males, the total incidence of acute myocardial infarction is 10% higher in Charleroi than in Ghent. In the female population, the incidence is 37% higher in the Walloon city compared to the Flemish city.

A controlled comparative study of chlorprothixene vs. haloperidol in chronic schizophrenia.

Neurochemical distinctions have been made between neuroleptic drugs that affect D-1 receptors as well as D-2 receptors, compared with those neuroleptic drugs that affect only D-2 receptors. However, a controlled double-blind study of haloperidol vs. chlorprothixene in schizophrenic patients found no significant differences.

Herpes simplex virus and cytomegalovirus in the serum of schizophrenic patients versus other psychosis and normal controls.

One hundred and thirty eight subjects were examined for presence of antibodies by immunoperidose assay (IPA). The frequency of Cytomegalovirus (CMV) and Herpes Simplex Virus (HSV) antibody of 48 schizophrenic inpatients was compared to 48 inpatients with other psychosis and to 48 staff controls. No differences were found between schizophrenic and control groups.

Respiratory and other symptoms in panic disorder versus other anxiety disorders.

Respiratory abnormalities have been proposed as a central feature of panic disorder, but the literature is not unanimous. Symptoms of anxiety were quantitatively recorded with the Hamilton Rating Scale for Anxiety (HAM-A) in 72 psychiatric out-patients in two anxiety disorders clinics in Israel; 44 patients had panic disorder with or without agoraphobia and 28 patients had other anxiety disorders. Panic patients had more cardiovascular symptoms, but not more respiratory symptoms, than other patients. The relative importance of respiratory symptoms in panic disorder is not yet settled.

Aggressiveness in subjects referred for examination by court order: a retrospective study (1988-90) in the southern district of Israel.

Two hundred and eighty cases examined by court order by the district psychiatry office in the southern district of Israel in the past three years (1988-90) were critically reviewed. Analysis of the monthly distribution of criminal acts throughout the year shows a specific pattern of monthly occurrence of aggressive criminal acts (annual rhythm). This rhythm shows peaks in February and the summer months (June, July, August) and nadirs in March, October, November and December. The monthly occurrence of non-aggressive criminal acts showed a different pattern as compared with the annual rhythm of aggressive criminal acts. A correlation was found between the annual rhythm of aggressive criminal acts and the annual photoperiod (day length) rhythm. There was a significant positive correlation between these rhythms. We also analysed the age dependent distribution of aggressive criminal acts. A continuous age dependent increase in the percentage rate of aggressiveness was found.

Aggressive behaviour as a cause of psychiatric admission: a comparison between schizophrenic and affective disorder patients.

One hundred and fifty-six subjects were randomly chosen from 1,560 patients admitted to two psychiatric hospitals in Israel during 1991. The subjects' files were retrospectively studied to determine whether aggressive behaviour was the reason for psychiatric admission. We examined the relationship between aggressive behaviour, major psychiatric disorders (schizophrenia and affective disorder) and demographic variables (sex, age, family status and education). We also studied the monthly distribution of aggression throughout the year and the correlation to daily photoperiod duration. Forty-six per cent of the admissions related to aggressive behaviour. Schizophrenic patients were overly represented in the aggressive group. The monthly distribution of aggressive behaviour differed between schizophrenic and affective disorder patients. While the latter showed a seasonal pattern of aggressiveness, with a statistically significant peak during spring and winter, schizophrenic aggressive patients were distributed equally throughout the year. No statistically significant correlation was found between the incidence of aggression and the photoperiod duration.