Therapie des neonatalen ARDS mit 1,2 - Dioleoyl-sn-glyzero-3-phospho-rac-(1-glycerol) Natrium-Salz

Das Projekt am neugeborenen Schweinemodell geht der Frage der bestmöglichen Therapie von akutem hypoxämischen Lungenversagen bei Neugeborenen nach. Das als nARDS (neonatal acute respiratory distress syndrome) bekannte Krankheitsbild ist eine komplexe Funktionsstörung des Gasaustausches und der Lungenmechanik als Folge einer schweren pulmonalen Inflammation, die eine hohe Sterblichkeit aufweist. Die Entzündung ist durch eine immunologische Reaktion gekennzeichnet, bei der u.a. das körpereigene Surfactant der Lunge durch Immunzellen und durch Plasmaproteine abgebaut wird. In diesem Versuch wurde bei maschinell beatmeten neugeborenen Ferkeln durch wiederholte Lungenlavagen mit physiologischer Kochsalzlösung über den Endotrachealtubus ein akutes Lungenversagen induziert. Im Verlauf einer Versuchsreihe von 72 Stunden wurde in Abständen von jeweils 24 Stunden die Lungen der Tiere zusätzlich mit zwei Phasen Lungen schädigender Beatmung sowie endotrachealer Instillation von Lipopolysacchariden geschädigt. Zwei Stunden nach jeder dieser Schädigungen erfolgte die therapeutische Intervention. Eine Gruppe wurde mit exogem endotracheal substituiertem Surfactant (S) behandelt, die Behandlungsgruppe bekam zur Surfactant-Emulsion das 1,2-Dioleoyl-sn-glyzero-3-phospho-rac-(1-glyzerol) Natrium-Salz (S+DOPG) zugesetzt und der Kontrollgruppe (C) wurde ein Luftbolus verabreicht. In der Gruppe mit S+DOPG stellten wir einige signifikante Verbesserungen der Parameter sowohl für den Gasaustausch, das Lungenödem, die Lungenmechanik als auch für die Ausprägung der Entzündungsreaktion fest. Die Oxygenierung, Ventilation, der extravaskuläre Lungenwasserindex und die spezifische Compliance des respiratorischen Systems waren signifikant besser. Außerdem waren die Genexpressionen von Amphiregulin, TGF-β1 und Interleukin-6 sowie die Konzentration von Phospholipase A2 signifikant verringert. Die Zellzahl in der bronchoalveolären Lavage und die Apoptoserate in den Lungenalveolen in der mit S+DOPG behandelten Gruppe waren signifikant verringert

Case report: HLA-haploidentical HSCT rescued with donor lymphocytes infusions in a patient with X-linked chronic granulomatous disease

Chronic granulomatous disease is an inborn error of immunity due to disrupted function of the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase complex. This results in impaired respiratory burst of phagocytes and insufficient killing of bacteria and fungi. Patients with chronic granulomatous disease are at increased risk for infections, autoinflammation and autoimmunity. Allogeneic hematopoietic stem cell transplantation (HSCT) is the only widely available curative therapy. While HSCT from human leukocyte antigen (HLA) matched siblings or unrelated donors are standard of care, transplantation from HLA-haploidentical donors or gene therapy are considered alternative options. We describe a 14-month-old male with X-linked chronic granulomatous disease who underwent a paternal HLA-haploidentical HSCT using T-cell receptor (TCR) alpha/beta(+)/CD19(+) depleted peripheral blood stem cells followed by mycophenolate graft versus host disease prophylaxis. Decreasing donor fraction of CD3(+) T cells was overcome by repeated infusions of donor lymphocytes from the paternal HLA-haploidentical donor. The patient achieved normalized respiratory burst and full donor chimerism. He remained disease-free off any antibiotic prophylaxis for more than three years after HLA-haploidentical HSCT. In patients with x-linked chronic granulomatous disease without a matched donor paternal HLA-haploidentical HSCT is a treatment option worth to consider. Administration of donor lymphocytes can prevent imminent graft failure

Acute mental stress drives vascular inflammation and promotes plaque destabilization in mouse atherosclerosis

Publisher Copyright: © 2021 The Author(s). Published by Oxford University Press on behalf of the European Society of Cardiology.Aims: Mental stress substantially contributes to the initiation and progression of human disease, including cardiovascular conditions. We aim to investigate the underlying mechanisms of these contributions since they remain largely unclear. Methods and results: Here, we show in humans and mice that leucocytes deplete rapidly from the blood after a single episode of acute mental stress. Using cell-tracking experiments in animal models of acute mental stress, we found that stress exposure leads to prompt uptake of inflammatory leucocytes from the blood to distinct tissues including heart, lung, skin, and, if present, atherosclerotic plaques. Mechanistically, we found that acute stress enhances leucocyte influx into mouse atherosclerotic plaques by modulating endothelial cells. Specifically, acute stress increases adhesion molecule expression and chemokine release through locally derived norepinephrine. Either chemical or surgical disruption of norepinephrine signalling diminished stress-induced leucocyte migration into mouse atherosclerotic plaques. Conclusion: Our data show that acute mental stress rapidly amplifies inflammatory leucocyte expansion inside mouse atherosclerotic lesions and promotes plaque vulnerability.publishersversionPeer reviewe

Three distinct pneumotypes characterize the microbiome of the lung in BALB/cJ mice.

Bacteria can rarely be isolated from normal healthy lungs using conventional culture techniques, supporting the traditional belief that the lungs are sterile. Yet recent studies using next generation sequencing report that bacterial DNA commonly found in the upper respiratory tract (URT) is present at lower levels in the lungs. Interpretation of that finding is complicated by the technical limitations and potential for contamination introduced when dealing with low biomass samples. The current work sought to overcome those limitations to clarify the number, type and source of bacteria present in the lungs of normal mice. Results showed that the oral microbiome is diverse and highly conserved whereas murine lung samples fall into three distinct patterns. 33% of the samples were sterile, as they lacked culturable bacteria and their bacterial DNA content did not differ from background. 9% of samples contained comparatively higher amounts of bacterial DNA whose composition mimicked that detected in the URT. A final group (58%) contained smaller amounts of microbial DNA whose composition was correlating to that of rodent chow and cage bedding, likely acquired by inspiration of food and bedding fragments. By analyzing each sample independently rather than working with group averages, this work eliminated the bias introduced by aspiration-contaminated samples to establish that three distinct microbiome pneumotypes are present in normal murine lungs

Three distinct pneumotypes characterize the microbiome of the lung in BALB/cJ mice

<div><p>Bacteria can rarely be isolated from normal healthy lungs using conventional culture techniques, supporting the traditional belief that the lungs are sterile. Yet recent studies using next generation sequencing report that bacterial DNA commonly found in the upper respiratory tract (URT) is present at lower levels in the lungs. Interpretation of that finding is complicated by the technical limitations and potential for contamination introduced when dealing with low biomass samples. The current work sought to overcome those limitations to clarify the number, type and source of bacteria present in the lungs of normal mice. Results showed that the oral microbiome is diverse and highly conserved whereas murine lung samples fall into three distinct patterns. 33% of the samples were sterile, as they lacked culturable bacteria and their bacterial DNA content did not differ from background. 9% of samples contained comparatively higher amounts of bacterial DNA whose composition mimicked that detected in the URT. A final group (58%) contained smaller amounts of microbial DNA whose composition was correlating to that of rodent chow and cage bedding, likely acquired by inspiration of food and bedding fragments. By analyzing each sample independently rather than working with group averages, this work eliminated the bias introduced by aspiration-contaminated samples to establish that three distinct microbiome pneumotypes are present in normal murine lungs.</p></div

The relative abundance of common URT and lung^unique genera.

<p>A: Differences between groups were determined by significant ANOVA using data from <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0180561#pone.0180561.g003" target="_blank">Fig 3</a>. Genera are shown and characterized by their significantly higher relative abundances in URT samples at an abundance of > 1%. B: The genera are shown as characterized by their significantly higher abundances in lung^unique samples with an abundance of > 1%. **; p < 0.01, ***; p < 0.001, ****; p < 0.0001, ns; not significant, Wilcox test.</p

Microbiome overlap between groups.

<p>Microbiome overlap between groups.</p

Heatmap.

<p>A heatmap was generated using the log₂ normalized number of sequences for genera with sample abundances >1%. The bacterial genera are shown in the rows and the individual samples are shown in the columns labeled by mouse of origin. Density of gray coloration reflects the relative frequency of each genus to the mean for that genus.</p

Group correlation coefficients.

<p>Group correlation coefficients.</p