Katharina Moritz

IL-32 is expressed by human primary keratinocytes and modulates keratinocyte apoptosis in atopic dermatitis

BACKGROUND Keratinocyte (KC) apoptosis is an important mechanism of eczema and spongiosis in patients with atopic dermatitis (AD) and is mediated by IFN-gamma, which is secreted by T(H)1 cells. IL-32 is a proinflammatory cytokine that is involved in the inflammatory processes of rheumatoid arthritis, chronic obstructive pulmonary disease, and Crohn disease. Recently, it was shown that upregulation of IL-32 induces apoptosis. OBJECTIVE The aim of the study was to investigate the expression and function of IL-32 in patients with AD. METHODS The expression of IL-32 in KCs was analyzed by means of RT-PCR, ELISA, and flow cytometry. Transfections of small interfering RNA were performed in primary KCs, and apoptosis was analyzed by means of terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling, annexin-V, and 7-amino actinomycin D stainings. Immunofluorescence stainings were used to detect IL-32 in skin biopsy specimens, and serum levels of IL-32 were analyzed by means of ELISA. RESULTS We report that IL-32 is expressed in human primary KCs on stimulation with IFN-gamma, TNF-alpha, and T(H)1 cells in contrast to T(H)2, regulatory T (Treg), or T(H)17 cells, which showed no effect. Transfection of primary KCs and artificial skin equivalents with small interfering RNA to IL-32, which resulted in a clear decrease in IL-32 expression, significantly reduced KC apoptosis. Immunofluorescence staining demonstrated that IL-32 was expressed in AD lesional skin, whereas it was present in neither skin biopsy specimens from healthy donors nor in lesional skin from patients with psoriasis. Serum levels of IL-32 from patients with AD correlated with disease severity, but increased serum levels of IL-32 were also detected in asthmatic patients. CONCLUSION The present study demonstrates KCs as a source of IL-32, which modulates KC apoptosis and contributes to the pathophysiology of AD.

TNF-like weak inducer of apoptosis (TWEAK) and TNF-α cooperate in the induction of keratinocyte apoptosis

BACKGROUND Activation of skin keratinocytes followed by their apoptotic death leads to eczema and spongiosis formations in patients with atopic dermatitis (AD). TNF-like weak inducer of apoptosis (TWEAK) binds to its receptor, fibroblast growth factor-inducible 14 (Fn14), and controls many cellular activities, including proliferation, migration, differentiation, apoptosis, angiogenesis, and inflammation. OBJECTIVE The aim of the study was to investigate the role of TWEAK and Fn14 in the formation of eczema in patients with AD. METHODS Primary keratinocytes were isolated from nonlesional skin from patients with AD and psoriasis and from normal skin of healthy donors. Apoptosis analysis was performed by using annexin V/7-aminoactinomycin D and terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling staining. The expression and regulation of TWEAK, TNF-α, Fn14, TNF receptor (TNFR) 1, and TNFR2 were measured by means of RT-PCR, flow cytometric analysis, and ELISA. TWEAK and Fn14 expression of lesional AD and psoriatic skin and normal control skin was analyzed by using immunohistochemistry and immunofluorescence. RESULTS TWEAK and TNF-α cooperate in the induction of apoptosis in primary keratinocytes obtained from patients with AD, patients with psoriasis, and healthy subjects and in artificial skin equivalents. TNFR1 and Fn14 were the main receptors involved. TWEAK upregulates TNF-α expression in primary keratinocytes, whereas TNF-α did not affect the expression of TWEAK and its receptors. High TWEAK expression was observed in AD lesions but not in psoriatic lesions or normal skin. Fn14 was highly expressed in the lesional skin of patients with AD and patients with psoriasis and in healthy control skin. CONCLUSION The high expression of TWEAK in lesional AD skin contributes to the difference in keratinocyte apoptosis and lesional formation between AD and psoriasis.

Frequent occurrence of T cell-mediated late reactions revealed by atopy patch testing with hypoallergenic rBet v 1 fragments.

Background Late allergic reactions are common in the course of allergen-specific immunotherapy and even occur with allergy vaccines with reduced IgE reactivity. Objective We sought to study atopy patch test (APT) reactions and T-cell responses to the recombinant birch pollen allergen Bet v 1 and recombinant hypoallergenic T-cell epitope–containing Bet v 1 fragments in patients with birch pollen allergy with and without atopic dermatitis (AD). Methods A clinical study was conducted in 15 patients with birch pollen allergy with AD (group 1), 5 patients with birch pollen allergy without AD (group 2), 5 allergic patients without birch pollen allergy (group 3), and 5 nonallergic subjects (group 4) by performing skin prick tests and APTs with rBet v 1 and hypoallergenic rBet v 1 fragments. T-cell, cutaneous lymphocyte antigen (CLA)+ and CCR4+ T-cell and cytokine responses were studied by thymidine uptake, carboxyfluorescein diacetate succinimidyl ester staining, and Luminex technology, respectively. Results rBet v 1 and hypoallergenic rBet v 1 fragments induced APT reactions in not only most of the patients with birch pollen allergy with AD (11/15) but also in most of those without AD (4/5). Patients with birch pollen allergy with AD had higher Bet v 1–specific proliferation of CLA+ and CCR4+ T cells compared with patients with birch pollen allergy without AD. There were no differences in Bet v 1–specific CLA+ and CCR4+ proliferation and cytokine secretion in patients with and without APT reactions. Conclusion Hypoallergenic rBet v 1 fragments induce T cell–dependent late reactions not only in patients with birch pollen allergy with AD but also in those without AD, which can be determined based on APT results but not based on in vitro parameters.

Malignancy and Specific Allergen Immunotherapy: The Results of a Case Series

Background: Specific immunotherapy with allergen is the only causative treatment for IgE-mediated allergies such as stinging insect allergy or hay fever and works by the induction of blocking antibodies and regulatory T lymphocytes. Objective: Does a hypothetical obstruction of tumor surveillance presupposing the induction of regulatory T cells really justify detaining immunotherapy to oncologic patients as suggested by recent guidelines? Methods: We report 6 patients (4 female, 2 male) suffering or having suffered from stage 1 cancer (4 melanomas, 1 lung cancer, 1 breast cancer) and concomitant IgE-mediated allergy. Four of them had a history of severe anaphylactic reactions to the insect yellow jacket, the 5th suffered from allergic rhinoconjunctivitis to dust mites, and the 6th to grass/rye pollen. Results: Between 2004 and 2010, subcutaneous immunotherapy was safely performed in 5 patients without signs of tumor reactivation. The cancer in 2 of them was diagnosed immediately after specific immunotherapy had been initiated and in another 2 the active cancer phase had already finished years before; the 5th suffered from a relapse around the time of the initiation of immunotherapy. At the time of the writing of the manuscript, 4 of them had already concluded 3 years of treatment, another one almost 1 year. The melanoma in the 6th patient was diagnosed 5 months after reaching the maintenance dose. Immunotherapy with grass/rye pollen was aborted in this patient based on current guidelines. Conclusions: Specific immunotherapy was safely administered in patients suffering concomitantly from IgE-mediated allergy and lower stage cancer.

Fixed drug eruption caused by mefenamic acid: a case series and diagnostic algorithms

Background: Fixed drug eruption is a fairly common drug‐induced hypersensitivity reaction of the skin and the mucous membranes, which is characterized by the re‐occurrence of the lesion(s) exactly on the previously involved sites after repeated administration. The pathogenetic mechanisms of this site‐specificity are not fully elucidated.

Epicutaneous allergen application preferentially boosts specific T cell responses in sensitized patients

The effects of epicutaneous allergen administration on systemic immune responses in allergic and non-allergic individuals has not been investigated with defined allergen molecules. We studied the effects of epicutaneous administration of rBet v 1 and rBet v 1 fragments on systemic immune responses in allergic and non-allergic subjects. We conducted a clinical trial in which rBet v 1 and two hypoallergenic rBet v 1 fragments were applied epicutaneously by atopy patch testing (APT) to 15 birch pollen (bp) allergic patients suffering from atopic dermatitis, 5 bp-allergic patients suffering from rhinoconjunctivitis only, 5 patients with respiratory allergy without bp allergy and 5 non-allergic individuals. Epicutaneous administration of rBet v 1 and rBet v 1 fragments led to strong and significant increases of allergen-specific T cell proliferation (CLA+ and CCR4+T cell responses) only in bp-allergic patients with a positive APT reaction. There were no relevant changes of Bet v 1-specific IgE and IgG responses. No changes were noted in allergic subjects without bp allergy and in non-allergic subjects. Epicutaneous allergen application boosts specific T cell but not antibody responses mainly in allergic, APT-positive patients suggesting IgE-facilitated allergen presentation as mechanism for its effects on systemic allergen-specific immune responses.

GA2LEN (Global Allergy and Asthma European Network), die Perspektive der deutschsprachigen Zentren

SummaryAllergic diseases represent a major health problem in Europe. They are increasing in prevalence, severity and costs. GA2LEN (Global Allergy and Asthma European Network), an FP6 Network of Excellence, was created in 2005 as a vehicle to ensure excellence in research bringing together research and clinical institutions to combat fragmentation in the European research area and to tackle allergy as a whole. GA2LEN benefited greatly from the voluntary efforts of researchers who are strongly committed to this model of pan-European collaboration. The network was organized in order to increase networking for scientific projects in allergy and asthma around Europe and to make GA2LEN the world leader in the field. Besides these activities, research has been jointly made and the first papers are being published. GA2LEN achievements in general can be grouped as those for a durable infrastructure built up during the project phase those which are project-related work based on these novel infrastructures, and the development and implementations of guidelines. The major achievements of GA2LEN are reported in this paper.ZusammenfassungAllergische Erkrankungen stellen eines der wesentlichen Gesundheitsprobleme in Europa dar. Häufigkeit, Schweregrad und Kosten dieser Erkrankungen weisen eine steigende Tendenz auf. 2005 wurde das GA2LEN Netzwerk (Global Allergy and Asthma European Network) gegründet, ein Exzellenz-Netzwerk des 6. EU Rahmenprogramms, das es sich zur Aufgabe gemacht hatte, Spitzenforschung sicherzustellen, indem es wissenschaftliche und klinische Einrichtungen vereinen sollte und dadurch die Aufsplitterung der europäischen Forschung überwinden könne. Allergien sollten als globales Problem in Angriff genommen werden. GA2LEN profitierte immens von den freiwilligen Anstrengungen der Wissenschaftler, die sich der Idee einer pan-europäischen Zusammenbarbeit verbunden fühlten. Ursprüngliches Ziel von GA2LEN war es gewesen, die Vernetzung wissenschaftlicher Projekte, die sich mit Allergien und Asthma in Europa beschäftigen, zu verstärken und das Netzwerk so an die weltweite Spitze der Allergie-Forschung zu setzen. Daneben wurden neue, gemeinsame Forschungsprojekte initiiert und gemeinsame Veröffentlichungen publiziert. Die Errungenschaften von GA2LEN lassen sich drei Untergruppen zuordnen: 1. den Aufbau einer tragfähigen Infrastruktur, 2. Projektbezogene Arbeit und 3. die Entwicklung und Implementierung von Leitlinien. In dieser Übersichtsarbeit werden die wichtigsten Errungenschaften des GA2LEN Netzwerks dargestellt.

Anaphylaxis to Malaysian meat loaf

e present the case of a 65-year-old woman, who presented erself at the allergy outpatient clinic because on her last oliday trip to Malaysia she had developed acute angioedema of the eyes, dysphagia and dyspnoea ten minutes fter the intake of a large breakfast, which consisted of crambled eggs and roasted potatoes, which had been tolrated before and after, and — for the first time — meat oaf. The hotel’s kitchen revealed the ingredients consistng in beef, potato flower, chicken stock and isolated soy rotein-soya bean paste but no peanut. The meat loaf had non-allergic subjects were used as negative controls. OD values were considered positive if they exceeded the mean OD of the negative controls by more than three standard deviations. For ELISA inhibitions experiments, individual patients’ sera were pre-incubated with birch pollen or soy protein extracts, rBet v 1 or rGly m 4 at final concentrations of 1, 10, 20 and 100 g/ml were added to the soy or birch pollen protein extract and to the rBet v 1 or rGly m 4-coated wells. IgE-binding was detected as described above. The skin prick test with commercial extracts showed positive results to birch pollen and the cross-reactive hazelnut, Candida albicans, dog, peanut, tomato, celery, but remained negative to soy extract. The total serum IgE was i ( w

GA2LEN (Global Allergy and Asthma European Network), die Perspektive der deutschsprachigen Zentren@@@GA2LEN (Global Allergy and Asthma European Network), the perspective of the German speaking centers

SummaryAllergic diseases represent a major health problem in Europe. They are increasing in prevalence, severity and costs. GA2LEN (Global Allergy and Asthma European Network), an FP6 Network of Excellence, was created in 2005 as a vehicle to ensure excellence in research bringing together research and clinical institutions to combat fragmentation in the European research area and to tackle allergy as a whole. GA2LEN benefited greatly from the voluntary efforts of researchers who are strongly committed to this model of pan-European collaboration. The network was organized in order to increase networking for scientific projects in allergy and asthma around Europe and to make GA2LEN the world leader in the field. Besides these activities, research has been jointly made and the first papers are being published. GA2LEN achievements in general can be grouped as those for a durable infrastructure built up during the project phase those which are project-related work based on these novel infrastructures, and the development and implementations of guidelines. The major achievements of GA2LEN are reported in this paper.ZusammenfassungAllergische Erkrankungen stellen eines der wesentlichen Gesundheitsprobleme in Europa dar. Häufigkeit, Schweregrad und Kosten dieser Erkrankungen weisen eine steigende Tendenz auf. 2005 wurde das GA2LEN Netzwerk (Global Allergy and Asthma European Network) gegründet, ein Exzellenz-Netzwerk des 6. EU Rahmenprogramms, das es sich zur Aufgabe gemacht hatte, Spitzenforschung sicherzustellen, indem es wissenschaftliche und klinische Einrichtungen vereinen sollte und dadurch die Aufsplitterung der europäischen Forschung überwinden könne. Allergien sollten als globales Problem in Angriff genommen werden. GA2LEN profitierte immens von den freiwilligen Anstrengungen der Wissenschaftler, die sich der Idee einer pan-europäischen Zusammenbarbeit verbunden fühlten. Ursprüngliches Ziel von GA2LEN war es gewesen, die Vernetzung wissenschaftlicher Projekte, die sich mit Allergien und Asthma in Europa beschäftigen, zu verstärken und das Netzwerk so an die weltweite Spitze der Allergie-Forschung zu setzen. Daneben wurden neue, gemeinsame Forschungsprojekte initiiert und gemeinsame Veröffentlichungen publiziert. Die Errungenschaften von GA2LEN lassen sich drei Untergruppen zuordnen: 1. den Aufbau einer tragfähigen Infrastruktur, 2. Projektbezogene Arbeit und 3. die Entwicklung und Implementierung von Leitlinien. In dieser Übersichtsarbeit werden die wichtigsten Errungenschaften des GA2LEN Netzwerks dargestellt.