Activation Patterns of Neutrophils in Familial Mediterranean Fever in Comparison to other acute and chronic inflammatory Diseases

Das Familiäre Mittelmeerfieber (FMF) zählt zu den häufigsten monogenen, autoinflammatorischen Erkrankungen und wird durch Gain-of-Function Mutationen im Familial Mediterranean Fever (MEFV)-Gen vererbt. Das MEFV Gen kodiert für das Protein Pyrin, das als Bestandteil des Inflammasoms in autoinflammatorischen Prozessen und in der Produktion von Interleukin-1β (IL-1β) involviert ist. Bislang galten autosomal rezessive Ergbänge als Auslöser der Erkrankung, mittlerweile gibt es Daten zu Krankheitssymptomen bei heterozygoten Mutationsträgern. Die Klinik der Erkrankung besteht in rezidivierenden Schüben, die in der Regel 24 - 72 Stunden anhalten und meist mit Fieberschüben einhergehen, häufig mit Peritonitis. Oft treten weitere Symptome wie akuter pleuritischer Schmerz, Arthritis im Bereich der Knie, Knöchel und Hüfte und erysipelähnliche Hautrötungen im Bereich des Unterschenkels auf. Die Diagnose wird häufig nach längerer, diagnostischer Vorgeschichte anhand der Klinik und des genetischen Screenings von Exon 1-10 des MEFV Gens gestellt. Allerdings ist das genetische Screening aufwändig und teuer und repräsentiert nicht die klinische Aktivität der Erkrankung. Ziel dieser Arbeit war es, ein spezifisches Aktivierungsmuster für das Familiäre Mittelmeerfieber zu beschreiben, welches sich auch für eine quantitative Evaluierung der Krankheitsaktivität des Patienten eignet. Ein weiteres Ziel war es zu zeigen, dass der Pathomechanismus im neutrophilen Granulozyten stattfindet, und dass es spezifische Aktivierungsmuster von Neutrophilen beim FMF in Abgrenzung zu anderen inflammatorischen Erkrankungen gibt. Zudem sollte untersucht werden, ob der dem Familiären Mittelmeerfieber zugrunde liegende Pathomechanismus durch Interleukin-1 gesteuert wird und vom NLRP3 Inflammasom abhängig ist. Aufbauend auf Vorarbeiten der Arbeitsgruppe PD Dr. Kallinich wurde hierzu die Sekretion von S100A12 Proteinen, Interleukin-18 und Caspase-1 in den aufgereinigten Neutrophilen von Patienten mit akuten und chronischen inflammatorischen Erkrankungen im zeitlichen Verlauf mit und ohne Stimulation sowie die Expression der Oberflächenmarker CD62L und CD11b untersucht. Die Ergebnisse wurden mit denen eines homozygoten und mit denen von heterozygot gesunden Mutationsträgern des Familiären Mittelmeerfiebers sowie mit Patienten mit anderen akuten und chronischen inflammatorischen Erkrankungen verglichen. Es zeigte sich ein krankheitsspezifisches Aktivierungsmuster. Mit den hier dargestellten Daten könnte zukünftig ein für die klinische Anwendung verfügbares diagnostisches Tool entwickelt werden. Darüber hinaus charakterisiert die vorliegende Arbeit die Rolle des angeborenen Immunsystems bei akuten und chronischen inflammatorischen Erkrankungen und beschreibt die phänotypischen Veränderungen von neutrophilen Granulozyten in sterilen und erregerbedingten Entzündungsprozessen.Familial Mediterranean Fever is one of the most common monogenic autoinflammatory diseases and is inherited by a gain of function mutation in the Familial Mediterranean Fever MEFV gene. The MEFV gene codes for the protein pyrin, which is a component of the inflammasome and is involved in autoinflammatory processes and in the production of interleukin-1β (IL-1β). Until recently, autosomal recessive findings were considered to be the trigger of the disease, but in the meantime data on disease symptoms in heterozygous mutation carriers are available. The clinical signs of the disease consist of recurrent attacks, which usually last 24-72 hours and are usually accompanied by fever attacks, often with peritonitis. Other symptoms often occur, such as acute pleuritic pain, arthritis in the knee, ankle and hip area, erysipelas-like reddening of the skin in the lower leg area. The diagnosis is often made after a long diagnostic history based on clinical and genetic screening of exons 1-10 of the MEFV gene. However, genetic screening is complex and expensive and does not represent the clinical activity of the disease. The aim of this work was to describe a specific activation pattern for Familial Mediterranean Fever which is also suitable for a quantitative evaluation of the disease activity of the patient. A further aim was to show that the pathomechanism takes place in neutrophil granulocytes, to present specific activation patterns of neutrophils in distinction to other inflammatory diseases and to investigate whether the pathomechanism underlying Familial Mediterranean Fever is controlled by interleukin-1 and is dependent on the NLRP3 inflammasome. Based on preliminary work of the research group PD Dr. Kallinich, the secretion of S100A12 proteins, interleukin-18 and caspase-1 in the purified neutrophils of patients with acute and chronic inflammatory diseases was investigated over time with and without stimulation as well as the shedding of the surface markers CD62L and CD11b. The results were compared with those of homozygous and heterozygous healthy mutation carriers of Familial Mediterranean Fever and with patients with other acute and chronic inflammatory diseases. A disease-specific activation pattern was found. With the data presented here, a diagnostic tool available for clinical application could be developed in the future. Furthermore, the present work characterizes the role of the innate immune system in acute and chronic inflammatory diseases and describes the phenotypic changes of neutrophil granulocytes in sterile and pathogen-induced inflammatory processes

Molecularly defined diffuse leptomeningeal glioneuronal tumor (DLGNT) comprises two subgroups with distinct clinical and genetic features

Diffuse leptomeningeal glioneuronal tumors (DLGNT) represent rare CNS neoplasms which have been included in the 2016 update of the WHO classification. The wide spectrum of histopathological and radiological features can make this enigmatic tumor entity difficult to diagnose. In recent years, large-scale genomic and epigenomic analyses have afforded insight into key genetic alterations occurring in multiple types of brain tumors and provide unbiased, complementary tools to improve diagnostic accuracy. Through genome-wide DNA methylation screening of > 25,000 tumors, we discovered a molecularly distinct class comprising 30 tumors, mostly diagnosed histologically as DLGNTs. Copy-number profiles derived from the methylation arrays revealed unifying characteristics, including loss of chromosomal arm 1p in all cases. Furthermore, this molecular DLGNT class can be subdivided into two subgroups [DLGNT methylation class (MC)-1 and DLGNT methylation class (MC)-2], with all DLGNT-MC-2 additionally displaying a gain of chromosomal arm 1q. Co-deletion of 1p/19q, commonly seen in IDH-mutant oligodendroglioma, was frequently observed in DLGNT, especially in DLGNT-MC-1 cases. Both subgroups also had recurrent genetic alterations leading to an aberrant MAPK/ERK pathway, with KIAA1549:BRAF fusion being the most frequent event. Other alterations included fusions of NTRK1/2/3 and TRIM33:RAF1, adding up to an MAPK/ERK pathway activation identified in 80% of cases. In the DLGNT-MC-1 group, age at diagnosis was significantly lower (median 5 vs 14 years, p < 0.01) and clinical course less aggressive (5-year OS 100, vs 43% in DLGNT-MC-2). Our study proposes an additional molecular layer to the current histopathological classification of DLGNT, of particular use for cases without typical morphological or radiological characteristics, such as diffuse growth and radiologic leptomeningeal dissemination. Recurrent 1p deletion and MAPK/ERK pathway activation represent diagnostic biomarkers and therapeutic targets, respectively—laying the foundation for future clinical trials with, e.g., MEK inhibitors that may improve the clinical outcome of patients with DLGNT

Gene-Dose Effect of MEFV Gain-of-Function Mutations Determines ex vivo Neutrophil Activation in Familial Mediterranean Fever

Familial Mediterranean fever (FMF) is caused by mutations within the Mediterranean fever (MEFV) gene. Disease severity depends on genotype and gene dose with most serious clinical courses observed in patients with M694V homozygosity. Neutrophils are thought to play an important role in the initiation and perpetuation of inflammatory processes in FMF, but little is known about the specific characteristics of these cells in FMF patients. To further characterize neutrophilic inflammatory responses in FMF and to delineate gene-dose effects on a cellular level, we analyzed cytokine production and activation levels of isolated neutrophils derived from patients and subjects with distinct MEFV genotypes, as well as healthy and disease controls. Serum levels of interleukin-18 (IL-18) (median 11,485 pg/ml), S100A12 (median 9,726 ng/ml), and caspase-1 (median 394 pg/ml) were significantly increased in patients with homozygous M694V mutations. Spontaneous release of S100A12, caspase-1, proteinase 3, and myeloperoxidase (MPO) was restricted to ex vivo cultured neutrophils derived from patients with two pathogenic MEFV mutations. IL-18 secretion was highest in patients with two mutations but also increased in neutrophils from healthy heterozygous MEFV mutation carriers, exhibiting an ex vivo gene-dose effect, which was formerly described by us in patients' serum. CD62L (l-selectin) was spontaneously shed from the surface of ex vivo cultured neutrophils [median of geometric mean fluorescence intensity (gMFI) after 5 h: 28.8% of the initial level]. While neutrophils derived from healthy heterozygous mutation carriers again showed a gene-dose effect (median gMFI: 67.1%), healthy and disease controls had significant lower shedding rates (median gMFI: 83.6 and 82.9%, respectively). Co-culture with colchicine and/or stimulation with adenosine triphosphate (ATP) and lipopolysaccharide (LPS) led to a significant increase in receptor shedding. Neutrophils were not prevented from spontaneous shedding by blocking IL-1 or the NLRP3 inflammasome. In summary, the data demonstrate that ex vivo cultured neutrophils derived from FMF patients display a unique phenotype with spontaneous release of high amounts of IL-18, S100A12, MPO, caspase-1, and proteinase 3 and spontaneous activation as demonstrated by the loss of CD62L. Neutrophilic activation seems to be independent from IL-1 activation and displays a gene-dose effect that may be responsible for genotype-dependent phenotypes

Encountering the archive in Katja Petrowskaja’s <i>Vielleicht Esther</i>

In Vielleicht Esther (2014), the literary debut by Ukrainian-born Katja Petrowskaja, the narrator attempts to trace her family history. She realizes that she can no longer rely on the memories of her relatives, but rather, as part of what Marianne Hirsch calls the "generation of postmemory," is dependent on the material that remains. She encounters various archive spaces and resources, but these fail to provide easy access to her family's past. This article argues that Vielleicht Esther is thus a pivotal example of an archival turn in memory culture, which signals not only the central position of the archive in retracing the past, but also the increasing critical scrutiny of the status and role of archive in this endeavour. Petrowskaja's narrator comes to see how the archive is implicated in the control of history and memory, and that what remains is also an indicator of what is missing - specifically the European Jewish tradition that once defined her ancestors. Moreover, the archive confronts her not only with what remains (and what doesn't), but also with questions about who remains (and why), that is, with questions about the circumstances of survival. On the one hand, her encounters with the archive allow her to address its gaps through narrative, but on the other, they confront her with unpalatable truths that force her to rethink her family narrative

A Pesquisa sobre Identidade e Cidadania nos EUA: da Nova História Social à Nova História Cultural

O presente artigo analisa as novas tendências da historiografia norte-americana a respeito da cidadania e da identidade, evidenciando os deslocamentos ocorridos tanto na escolha dos objetos de pesquisa quanto na abordagem. A passagem da história social para a história cultural implicou em rupturas significativas e abriu novas direções de pesquisa, perceptível na produção mais recente.<br>The present article deals with the new tendencies of north american historiography about citizenship and identity, emphasizing the birth of different choices on research subjects and approaches. The move from social history to cultural history meant important ruptures and opened new directions of research which are visible in the most recent literature