A workshop report on the FDNext project funded by the German Research Foundation

Nach zwei Jahren Projektlaufzeit lud der DFG-geförderte Projektverbund FDNext zu einem zweiten Community-Workshop ein. Unter dem Motto „Nachhaltiges Forschungsdatenmanagement gemeinsam umsetzen“ wurde eine projektweite Ergebnisbilanz gezogen und im Rahmen einer Online-Veranstaltung vorgestellt. Einzelne Formate ermöglichten den Austausch und die Diskussion zur Vision des Kulturwandels und eines ganzheitlichen FDMs durch Initiativen wie die Nationale Forschungsdateninfrastruktur (NFDI) sowie die Möglichkeiten der Zusammenarbeit zwischen einzelnen Konsortien und Hochschulen. Dabei wurden Aufgaben identifiziert, welche nur gemeinsam mit der FDM- bzw. Wissenschafts-Community bearbeitet werden können.Two years into the project duration, the collaborative project FDNext convened its second community workshop titled “Implementing Sustainable Research Data Management in a Joint Project”. Focusing on a review of achievements, the online event presented findings from all participating parties. Various formats fostered exchange and debates about perspectives of cultural change and a holistic research data management through initiatives such as the Nationale Forschungsdateninfrastruktur NFDI (national research data infrastructure), as well as collaboration opportunities between individual consortia and universities. Tasks and challenges that can only be dealt with in cooperation with RDM and scientific communities have been identified.Peer Reviewe

Two monoclonal antibodies against glycoprotein Gn protect mice from Rift Valley Fever challenge by cooperative effects.

Rift Valley fever virus (RVFV) is a zoonotic arbovirus that causes severe disease in humans and ruminants. The infection is characterized by abortions in pregnant animals, high mortality in neonates as well as febrile illness in humans that develop in 1% of cases encephalitis or hemorrhagic fever. There is presently no specific antiviral treatment for RVFV infection available. In this study, two monoclonal antibodies (mAbs), raised against glycoprotein Gn, were applied in a therapeutic study. Treatment of RVFV infected mice with neutralizing mAb Gn3 alone at two different time points (30 minutes before or 30 minutes after virus challenge) showed only moderate efficacy of about 58.3% survival in both applications. However, a combination therapy together with non-neutralizing mAb Gn32 demonstrated complete protection (100% survival) when applied 30 minutes after the lethal challenge dose. The increase of mAb efficacy is probably based on cooperative neutralization effects. These data suggest that a combination therapy with mAbs Gn3 and Gn32 could be an effective treatment option against RVFV infection

Gut Microbial Colonization Orchestrates TLR2 Expression, Signaling and Epithelial Proliferation in the Small Intestinal Mucosa

<div><p>The gut microbiota is an environmental factor that determines renewal of the intestinal epithelium and remodeling of the intestinal mucosa. At present, it is not resolved if components of the gut microbiota can augment innate immune sensing in the intestinal epithelium via the up-regulation of Toll-like receptors (TLRs). Here, we report that colonization of germ-free (GF) Swiss Webster mice with a complex gut microbiota augments expression of TLR2. The microbiota-dependent up-regulation of components of the TLR2 signaling complex could be reversed by a 7 day broad-spectrum antibiotic treatment. TLR2 downstream signaling via the mitogen-activated protein kinase (ERK1/2) and protein-kinase B (AKT) induced by bacterial TLR2 agonists resulted in increased proliferation of the small intestinal epithelial cell line MODE-K. Mice that were colonized from birth with a normal gut microbiota (conventionally-raised; CONV-R) showed signs of increased small intestinal renewal and apoptosis compared with GF controls as indicated by elevated mRNA levels of the proliferation markers Ki67 and Cyclin D1, elevated transcripts of the apoptosis marker Caspase-3 and increased numbers of TUNEL-positive cells per intestinal villus structure. In accordance, TLR2-deficient mice showed reduced proliferation and reduced apoptosis. Our findings suggest that a tuned proliferation response of epithelial cells following microbial colonization could aid to protect the host from its microbial colonizers and increase intestinal surface area.</p></div

Germ-free housing conditions do not affect aortic root and aortic arch lesion size of late atherosclerotic low-density lipoprotein receptor-deficient mice.

The microbiota has been linked to the development of atherosclerosis, but the functional impact of these resident bacteria on the lesion size and cellular composition of atherosclerotic plaques in the aorta has never been experimentally addressed with the germ-free low-density lipoprotein receptor-deficient (Ldlr-/- ) mouse atherosclerosis model. Here, we report that 16 weeks of high-fat diet (HFD) feeding of hypercholesterolemic Ldlr-/- mice at germ-free (GF) housing conditions did not impact relative aortic root plaque size, macrophage content, and necrotic core area. Likewise, we did not find changes in the relative aortic arch lesion size. However, late atherosclerotic GF Ldlr-/- mice had altered inflammatory plasma protein markers and reduced smooth muscle cell content in their atherosclerotic root plaques relative to CONV-R Ldlr-/- mice. Neither absolute nor relative aortic root or aortic arch plaque size correlated with age. Our analyses on GF Ldlr-/- mice did not reveal a significant contribution of the microbiota in late aortic atherosclerosis

TLR2 signals in the small intestinal epithelium evoke a proliferation response and increase signs of apoptosis in terminally differentiated epithelium.

<p><b>a–c</b>, Relative Ki67, Cyclin D1 or Caspase-3 mRNA levels in small intestine from GF and CONV-R mice (n = 7 Swiss Webster mice per group). <b>d–f</b>, Relative Ki67, Cyclin D1 or Caspase-3 mRNA levels in small intestine from Tlr2^−/− mice compared with WT controls (n = 7 C57BL/6J mice per group). <b>g</b>, Number of Caspase-3 positive cells per 10 villi in small intestinal samples from WT or Tlr2^−/− mice. <b>h–j</b>, Relative Ki67, Cyclin D1 and Caspase-3 mRNA levels in small intestine from mice colonized for 14 days with <i>E. coli</i> JP313 (n = 7 C57BL/6 mice per group). <b>k</b>, Relative Ki67 mRNA levels in MODE-K cells stimulated with PG (50 µg/ml) for 2, 4, or 8 hours (n = 4). <b>l</b>, Relative proliferation of PG treated MODE-K cells measured by incorporation of BrdU compared to untreated control cells (n = 3). <b>m</b>, Relative proliferation of MODE-K cells transfected with siRNA against TLR2 or scrambled control RNA measured by incorporation of BrdU (n = 4). <b>n</b>, TLR2 immunoblot of siRNA transfected MODE-K cells. Results are shown as means ±s.e.m. One asterisk, P<0.05; two asterisks, P<0.01; three asterisks, P<0.005; four asterisks, P<0.001.</p

Adapter molecules MyD88 and TRIF alter expression of TLRs in the ileum – indications for TLR receptor cross-talk in the small intestine.

<p><b>a+b</b>, Relative MyD88 and TRIF mRNA levels in small intestine from GF, CONV-D and CONV-R mice (n = 6–7 Swiss Webster mice per group). <b>c+d</b>, Relative MyD88 and TRIF mRNA levels in small intestine from CONV-R mice treated for 7 days with an antibiotic cocktail (ABX) (n = 6–8 Swiss Webster mice per group). <b>e–h</b>, Relative TLR2, 1, 6 and 4 mRNA levels in small intestine from MyD88^−/− mice compared to wildtype (WT) controls (n = 6–7 C57BL/6J mice per group). <b>i–l</b>, Relative TLR2, 1, 6 and 4 mRNA levels in small intestine from TRIF^−/− mice compared to WT controls (n = 6–7 mice per group). <b>m–o</b>, Relative TLR1, 6 and 4 mRNA levels in small intestine from TLR2^−/− mice compared to WT controls (n = 6–7 mice per group). <b>p</b>, Relative TLR2 mRNA levels in small intestine from TLR4^−/− mice compared to WT controls (n = 6–7 mice per group). Female mice were analyzed. Results are shown as means ± s.e.m. One asterisk, P<0.05; two asterisks, P<0.01; three asterisks, P<0.005; four asterisks, P<0.001.</p

Monocolonization with <i>E. coli</i> JP313 decreases TLR6 transcript levels.

<p><b>a–d,</b> Relative TLR2, 1, 6, and 4 mRNA levels in small intestine from mice colonized for 14 days with <i>E. coli</i> JP313 (n = 7 male C57BL/6 mice per group). Results are shown as means ± s.e.m. Two asterisks, P<0.01.</p

Microbial colonization leads to induction of TLR2 receptor expression in the small intestine.

<p><b>a,</b> Relative TLR2 mRNA levels in small intestinal tissues from GF, CONV-D and CONV-R mice (n = 7 Swiss Webster mice per group). <b>b,</b> TLR2 immunoblot of isolated small-intestinal enterocyte lysates from GF and CONV-R mice (n = 6 mice per group; shown is one representative blot). <b>c–e,</b> Relative TLR1, 6 and 4 mRNA levels in small intestinal tissues from GF, CONV-D and CONV-R mice (n = 6–7 mice per group). <b>f,</b> qPCR analyses of feces samples of control mice and mice treated with antibiotics for 7 days. qPCR was performed using universal primers for 16S bacterial sequences and normed to <i>E. coli</i> bacterial counts (cfu/µl). <b>g–j,</b> Relative mRNA levels of TLR2, 1, 6 and 4 in small intestinal tissues from CONV-R mice treated with a cocktail (ABX) of ampicillin (1 g/L) and neomycin (0.5 g/L) for 7 days (n = 6–7 mice per group). Female Swiss Webster mice or cells isolated from these mice were analyzed. Results are shown as means ± s.e.m. One asterisk, P<0.05; two asterisks, P<0.01; three asterisks, P<0.005.</p

Agonist-specific orchestration of the TLR expression profile in a mouse small intestinal epithelial cell line.

<p><b>a+b,</b> Relative mRNA expression of TLR2 in MODE-K cells stimulated with PG (50 µg/ml) or Pam3CSK4 (0.5 µg/ml) for 2, 4 or, 8 hours (n = 4). <b>c,</b> TLR2 immunoblot of PG or Pam3CSK4 stimulated MODE-K cells. Cells were treated with or without (CTR) PG or Pam3CSK4 for 2, 4, or 8 hours (n = 3, representative blot). <b>d–f,</b> Relative TLR1, 6 and 4 mRNA expression in MODE-K cells stimulated with Pam3CSK4 or LPS for 2, 4, or 8 hours (n = 4). Results are shown as means ± s.e.m. One asterisk, P<0.05; two asterisks, P<0.01; three asterisks, P<0.005; four asterisks, P<0.001.</p

Primer Table.

<p>Primer Table.</p