Studien zur Aufklärung der immunosuppressiven Wirkweise von mesenchymalen Stammzellen in Kombinationstherapie mit Mycophenolat-Mofetil

Mesenchymale Stammzellen haben sich in den letzten Jahren immer mehr zu einer vielversprechenden Möglichkeit bei der Therapie verschiedenster Krankheitsbilder entwickelt. In der Transplantationsmedizin bieten sie sich als komplementäre Therapieform zu den etablierten, immunsuppressiven Therapien an und eröffnen neue Möglichkeiten. Am erfolgreichsten werden MSCs heute schon in der Therapie der Graft-versus-Host-Disease eingesetzt. Bei unseren Untersuchungen stellte sich heraus, dass durch die Verabreichung mesenchymaler Stammzellen in Kombinationstherapie mit MMF bei der Transplantation solider Organe, ein Vorteil, bezogen auf das Transplantatüberleben im Tiermodell zu erreichen ist. Ein entscheidender Mechanismus für diese Beobachtung könnte sein, dass durch die am Tag -4 vor der Transplantation in die Schwanzvene injizierten MSC, T-Zellen zwar vorsensibilisiert werden, diese jedoch durch die 7-tägige Gabe von 20mg/kgKG Mycophenolat-Mofetil wieder eliminiert werden. Die frühe Aktivierung der APCs wird durch einen hohen Anteil aktivierter APCs (Ox6+/CD86+) an Tag 1 in abdominalen Lymphknoten als auch in der Milz beschrieben. Dies konnte in den Untersuchungen zum einen der CD4+/CD28+ T-Zellen in abdominellen Lymphknoten sowie den CD8+/CD28+ T-Zellen der Milz, in denen eine deutliche Expression von Aktivierungsmarkern durchflusszytometrisch vorlag, als auch durch die ausbleibende Migration der APCs in das Transplantat der Versuchsgruppe, immunhistochemisch gezeigt werden. Zudem spricht die verminderte Anzahl CD8+/CD28+ T-Zellen an Tag 10 im Vergleich von MMF und MSC behandelten Tieren zu den Tieren der Kontrollgruppe für These der Elimination durch MMF. Auch der prozentuale Anteil der Ox6+/CD86+ Zellen im peripheren Blut und der Milz an Tag 1 der Untersuchungen unterstützt dies. Nicht zuletzt benötigen mesenchymale Stammzellen eine Umgebung der Zytokinstimulation um effektiv immunsuppressiv wirken zu können. In den im Versuchsaufbau gemessene INF-γ Level im Serum der Versuchstiere aller Gruppen konnten keine Unterschiede festgestellt werden, sodass davon ausgegangen werden kann, dass eine effektive Aktivierung der MSC möglich ist und keine Beeinflussung durch MMF vorliegt. In der Zusammenschau der Ergebnisse ergibt sich das Bild, dass der Erfolg der Kombinationstherapie von MSC und MMF auf mehreren Standbeinen fußt. Einerseits erfolgt eine Modulation des Endothels der transplantierten Organe. Damit wird eine verringerte Transmigation der APCs und Leukozyten in das Transplantat erreicht. Andererseits werden aktivierte Lymphozyten durch die Verabreichung von MMF eliminiert. Gleichzeitig werden Ausreifung und Differenzierung der antigenpräsentierenden Zellen verzögert, aber nicht verhindert. Auch unter diesem Aspekt ist eine mögliche Rolle regulatorischer APCs denkbar

A cohort of 17 patients with kyphoscoliotic Ehlers-Danlos syndrome caused by biallelic mutations in FKBP14: expansion of the clinical and mutational spectrum and description of the natural history.

PurposeIn 2012 we reported in six individuals a clinical condition almost indistinguishable from PLOD1-kyphoscoliotic Ehlers-Danlos syndrome (PLOD1-kEDS), caused by biallelic mutations in FKBP14, and characterized by progressive kyphoscoliosis, myopathy, and hearing loss in addition to connective tissue abnormalities such as joint hypermobility and hyperelastic skin. FKBP14 is an ER-resident protein belonging to the family of FK506-binding peptidyl-prolyl cis-trans isomerases (PPIases); it catalyzes the folding of type III collagen and interacts with type III, type VI, and type X collagens. Only nine affected individuals have been reported to date.MethodsWe report on a cohort of 17 individuals with FKBP14-kEDS and the follow-up of three previously reported patients, and provide an extensive overview of the disorder and its natural history based on clinical, biochemical, and molecular genetics data.ResultsBased on the frequency of the clinical features of 23 patients from the present and previous cohorts, we define major and minor features of FKBP14-kEDS. We show that myopathy is confirmed by histology and muscle imaging only in some patients, and that hearing impairment is predominantly sensorineural and may not be present in all individuals.ConclusionOur data further support the extensive clinical overlap with PLOD1-kEDS and show that vascular complications are rare manifestations of FKBP14-kEDS

The German National Registry of Primary Immunodeficiencies (2012-2017)

Introduction: The German PID-NET registry was founded in 2009, serving as the first national registry of patients with primary immunodeficiencies (PID) in Germany. It is part of the European Society for Immunodeficiencies (ESID) registry. The primary purpose of the registry is to gather data on the epidemiology, diagnostic delay, diagnosis, and treatment of PIDs. Methods: Clinical and laboratory data was collected from 2,453 patients from 36 German PID centres in an online registry. Data was analysed with the software Stata® and Excel. Results: The minimum prevalence of PID in Germany is 2.72 per 100,000 inhabitants. Among patients aged 1–25, there was a clear predominance of males. The median age of living patients ranged between 7 and 40 years, depending on the respective PID. Predominantly antibody disorders were the most prevalent group with 57% of all 2,453 PID patients (including 728 CVID patients). A gene defect was identified in 36% of patients. Familial cases were observed in 21% of patients. The age of onset for presenting symptoms ranged from birth to late adulthood (range 0–88 years). Presenting symptoms comprised infections (74%) and immune dysregulation (22%). Ninety-three patients were diagnosed without prior clinical symptoms. Regarding the general and clinical diagnostic delay, no PID had undergone a slight decrease within the last decade. However, both, SCID and hyper IgE- syndrome showed a substantial improvement in shortening the time between onset of symptoms and genetic diagnosis. Regarding treatment, 49% of all patients received immunoglobulin G (IgG) substitution (70%—subcutaneous; 29%—intravenous; 1%—unknown). Three-hundred patients underwent at least one hematopoietic stem cell transplantation (HSCT). Five patients had gene therapy. Conclusion: The German PID-NET registry is a precious tool for physicians, researchers, the pharmaceutical industry, politicians, and ultimately the patients, for whom the outcomes will eventually lead to a more timely diagnosis and better treatment

Suitable Disinfectants with Proven Efficacy for Genetically Modified Viruses and Viral Vectors

Viral disinfection is important for medical facilities, the food industry, and the veterinary field, especially in terms of controlling virus outbreaks. Therefore, standardized methods and activity levels are available for these areas. Usually, disinfectants used in these areas are characterized by their activity against test organisms (i.e., viruses, bacteria, and/or yeasts). This activity is usually determined using a suspension test in which the test organism is incubated with the respective disinfectant in solution to assess its bactericidal, yeasticidal, or virucidal activity. In addition, carrier methods that more closely reflect real-world applications have been developed, in which microorganisms are applied to the surface of a carrier (e.g., stainless steel frosted glass, or polyvinyl chloride (PVC)) and then dried. However, to date, no standardized methods have become available for addressing genetically modified vectors or disinfection-resistant oncolytic viruses such as the H1-parvovirus. Particularly, such non-enveloped viruses, which are highly resistant to disinfectants, are not taken into account in European standards. This article proposes a new activity claim known as “virucidal activity PLUS”, summarizes the available methods for evaluating the virucidal activity of chemical disinfectants against genetically modified organisms (GMOs) using current European standards, including the activity against highly resistant parvoviridae such as the adeno-associated virus (AAV), and provides guidance on the selection of disinfectants for pharmaceutical manufacturers, laboratories, and clinical users

Volume-Assisted Estimation of Remnant Liver Function Based on Gd-EOB-DTPA Enhanced MR Relaxometry: A Prospective Observational Trial

In the context of liver surgery, predicting postoperative liver dysfunction is essential. This study explored the potential of preoperative liver function assessment by MRI for predicting postoperative liver dysfunction and compared these results with the established indocyanine green (ICG) clearance test. This prospective study included patients undergoing liver resection with preoperative MRI planning. Liver function was quantified using T1 relaxometry and correlated with established liver function scores. The analysis revealed an improved model for predicting postoperative liver dysfunction, exhibiting an accuracy (ACC) of 0.79, surpassing the 0.70 of the preoperative ICG test, alongside a higher area under the curve (0.75). Notably, the proposed model also successfully predicted all cases of liver failure and showed potential in predicting liver synthesis dysfunction (ACC 0.78). This model showed promise in patient survival rates with a Hazard ratio of 0.87, underscoring its potential as a valuable tool for preoperative evaluation. The findings imply that MRI-based assessment of liver function can provide significant benefits in the early identification and management of patients at risk for postoperative liver dysfunction

Gerodermia osteodysplastica is caused by mutations in SCYL1BP1, a Rab-6 interacting golgin

Gerodermia osteodysplastica is an autosomal recessive disorder characterized by wrinkly skin and osteoporosis. Here we demonstrate that gerodermia osteodysplastica is caused by loss-of-function mutations in SCYL1BP1, which is highly expressed in skin and osteoblasts. The protein localizes to the Golgi apparatus and interacts with Rab6, identifying SCYL1BP1 as a golgin. These results associate abnormalities of the secretory pathway with age-related changes in connective tissues

Clinical pattern, mutations and in vitro residual activity in 33 patients with severe 5, 10 methylenetetrahydrofolate reductase (MTHFR) deficiency

Background Severe methylenetetrahydrofolate reductase (MTHFR) deficiency is a rare inborn defect disturbing the remethylation of homocysteine to methionine ( Methods Clinical, biochemical and treatment data was obtained from physicians by using a questionnaire. MTHFR activity was measured in primary fibroblasts; genomic DNA was extracted from cultured fibroblasts. Results Thirty-three patients (mean age at follow-up 11.4 years; four deceased; median age at first presentation 5 weeks; 17 females) were included. Patients with very low (1.7-34.8 %) residual enzyme activity had mainly psychiatric symptoms, mental retardation, myelopathy, ataxia and spasticity. Treatment with various combinations of betaine, methionine, folate and cobalamin improved the biochemical and clinical phenotype. During the disease course, patients with very low enzyme activity showed a progression of feeding problems, neurological symptoms, mental retardation, and psychiatric disease while in patients with higher residual enzyme activity, myelopathy, ataxia and spasticity increased. All other symptoms remained stable or improved in both groups upon treatment as did brain imaging in some cases. No clear genotype-phenotype correlation was obvious. Discussion MTHFR deficiency is a severe disease primarily affecting the central nervous system. Age at presentation and clinical pattern are correlated with residual enzyme activity. Treatment alleviates biochemical abnormalities and clinical symptoms partially