R-esp1, a rat homologue of Drosophila Groucho, is differentially expressed after optic nerve crush and mediates NGF-induced survival of PC12 cells

AbstractThe differential display reverse transcription polymerase chain reaction method was used to detect alterations in gene expression in the superior colliculus after optic nerve crush in adult rats. One of the most prominent changes observed was the selective induction of R-esp1, a homologue of the Drosophila enhancer of split locus (Groucho). Therefore, we studied the influence of R-esp1 on nerve growth factor (NGF)-induced cell survival of PC12 cells. Overexpression of R-esp1 promotes cell survival even in the absence of NGF and, conversely, it is reduced by antisense-mediated inhibition of R-esp1 expression. In conclusion, we propose a novel model in which R-esp1 protein mediates the NGF-signaling pathway

KLINISCHE UND PATHOPHYSIOLOGISCHE ASPEKTE ENTZÜNDLICHER NIERENGEFÄSSERKRANKUNGEN

Die primären Entzündungen der Blutgefäße (primäre systemische Vaskulitiden) gehören zu den Autoimmunerkrankungen. Hierunter werden Erkrankungen verstanden, bei denen Strukturen des eigenen Organismus durch immunkompetente Zellen oder Autoantikörper angegriffen werden. Innerhalb dieser komplexen Gruppe verschiedener Erkrankungen sind die Entzündungen der kleinen Gefäße, zu denen auch die Kapillarschlingen in den Glomeruli (Nierenkörperchen) gehören, für die Nierenfunktion von erheblicher Bedeutung, da die Wände der glomerulären Kapillaren die Filtrationsbarriere in der Niere darstellen. Vaskulitiden, welche die kleinen Gefäße der Niere betreffen, können daher zu einem raschen Nierenfunktionsverlust mit der Konsequenz einer Dialysepflichtigkeit führen. Darüber hinaus können Beteiligungen anderer Organsysteme, vor allem der Lunge, mit einem akut lebensbedrohlichen Krankheitsbild einhergehen. Die schnelle Diagnosestellung dieser Erkrankungen ist für die Prognose der betroffenen Patienten entscheidend, um durch eine frühzeitige Therapieeinleitung einerseits lebensbedrohliche Folgen und andererseits ein dialysepflichtiges Nierenversagen zu verhindern. Die vorliegende Arbeit soll verschiedene Formen der Vaskulitiden mit Nierenbeteiligung beschreiben, die aktuelle Diagnostik und Therapie aufzeigen und Teilaspekte zur Pathophysiologie, mit denen sich unsere Arbeitsgruppe in den letzten Jahren beschäftigt hat, diskutieren

Characterization of Maladaptive Processes in Acute, Chronic and Remission Phases of Experimental Colitis in C57BL/6 Mice

Inflammatory bowel disease (IBD) is a chronic recurrent inflammatory disease with unknown etiology. Dextran sulfate sodium (DSS) induced colitis is a widely used mouse model in IBD research. DSS colitis involves activation of the submucosal immune system and can be used to study IBD-like disease characteristics in acute, chronic, remission and transition phases. Insight into colon inflammatory parameters is needed to understand potentially irreversible adaptations to the chronification of colitis, determining the baseline and impact of further inflammatory episodes. We performed analyses of non-invasive and invasive colitis parameters in acute, chronic and remission phases of the DSS colitis in C57BL/6 mice. Non-invasive colitis parameters poorly reflected inflammatory aspects of colitis in chronic remission phase. We found invasive inflammatory parameters, positively linked to repeated DSS-episodes, such as specific colon weight, inflamed colon area, spleen weight, absolute cell numbers of CD4+ and CD8+ T cells as well as B cells, blood IFN-γ level, colonic chemokines BLC and MDC as well as the prevalence of Turicibacter species in feces. Moreover, microbial Lactobacillus species decreased with chronification of disease. Our data point out indicative parameters of recurrent gut inflammation in context of DSS colitis

Inhibitors of Dipeptidyl Peptidase IV and Aminopeptidase N Target Major Pathogenetic Steps in Acne Initiation

Acne is a chronic disease hallmarked by sebaceous hyperplasia, follicular hyperkeratosis, and inflammation. Parallel targeting of these factors is required to treat acne effectively. Inhibitors of dipeptidyl peptidase IV (DP IV) and aminopeptidase N (APN) show strong anti-inflammatory effects on immune cells and therapeutic efficacy in autoimmune disorders. Our investigation focused on the expression and functional relevance of these ectopeptidases in three cell types which exhibit an altered phenotype in early acne lesions. We showed for the first time expression of DP IV and APN on human sebocytes. In the SZ95 sebocyte cell line, the DP IV inhibitors Lys[Z(NO2)]-thiazolidide and Lys[Z(NO2)]-pyrrolidide and the APN inhibitors actinonin and bestatin suppressed proliferation, enhanced terminal differentiation, and slightly decreased total neutral lipid production. The anti-inflammatory and differentiation-restoring cytokine IL-1 receptor antagonist was significantly upregulated in SZ95 sebocytes and the HaCaT keratinocyte cell line in the presence of inhibitors. Furthermore, the inhibitors suppressed proliferation and IL-2 production of Propionibacterium acnes-stimulated T cells ex vivo and enhanced the expression of the immunosuppressive cytokine transforming growth factor-β1. Our data provide first evidence for a functional role of DP IV and APN in the sebaceous gland apparatus and for their inhibitors, used alone or in combination, as completely new substances possibly affecting acne pathogenesis in a therapeutic manner

The Changing Landscape for Stroke\ua0Prevention in AF: Findings From the GLORIA-AF Registry Phase 2

Background GLORIA-AF (Global Registry on Long-Term Oral Antithrombotic Treatment in Patients with Atrial Fibrillation) is a prospective, global registry program describing antithrombotic treatment patterns in patients with newly diagnosed nonvalvular atrial fibrillation at risk of stroke. Phase 2 began when dabigatran, the first non\u2013vitamin K antagonist oral anticoagulant (NOAC), became available. Objectives This study sought to describe phase 2 baseline data and compare these with the pre-NOAC era collected during phase 1. Methods During phase 2, 15,641 consenting patients were enrolled (November 2011 to December 2014); 15,092 were eligible. This pre-specified cross-sectional analysis describes eligible patients\u2019 baseline characteristics. Atrial fibrillation disease characteristics, medical outcomes, and concomitant diseases and medications were collected. Data were analyzed using descriptive statistics. Results Of the total patients, 45.5% were female; median age was 71 (interquartile range: 64, 78) years. Patients were from Europe (47.1%), North America (22.5%), Asia (20.3%), Latin America (6.0%), and the Middle East/Africa (4.0%). Most had high stroke risk (CHA2DS2-VASc [Congestive heart failure, Hypertension, Age  6575 years, Diabetes mellitus, previous Stroke, Vascular disease, Age 65 to 74 years, Sex category] score  652; 86.1%); 13.9% had moderate risk (CHA2DS2-VASc = 1). Overall, 79.9% received oral anticoagulants, of whom 47.6% received NOAC and 32.3% vitamin K antagonists (VKA); 12.1% received antiplatelet agents; 7.8% received no antithrombotic treatment. For comparison, the proportion of phase 1 patients (of N = 1,063 all eligible) prescribed VKA was 32.8%, acetylsalicylic acid 41.7%, and no therapy 20.2%. In Europe in phase 2, treatment with NOAC was more common than VKA (52.3% and 37.8%, respectively); 6.0% of patients received antiplatelet treatment; and 3.8% received no antithrombotic treatment. In North America, 52.1%, 26.2%, and 14.0% of patients received NOAC, VKA, and antiplatelet drugs, respectively; 7.5% received no antithrombotic treatment. NOAC use was less common in Asia (27.7%), where 27.5% of patients received VKA, 25.0% antiplatelet drugs, and 19.8% no antithrombotic treatment. Conclusions The baseline data from GLORIA-AF phase 2 demonstrate that in newly diagnosed nonvalvular atrial fibrillation patients, NOAC have been highly adopted into practice, becoming more frequently prescribed than VKA in Europe and North America. Worldwide, however, a large proportion of patients remain undertreated, particularly in Asia and North America. (Global Registry on Long-Term Oral Antithrombotic Treatment in Patients With Atrial Fibrillation [GLORIA-AF]; NCT01468701

Cutting Edge: Innate Lymphoid Cells Suppress Homeostatic T Cell Expansion in Neonatal Mice

In adult mice, lymphopenia-induced proliferation (LIP) leads to T cell activation, memory differentiation, tissue destruction, and a loss of TCR diversity. Neonatal mice are lymphopenic within the first week of life. This enables some recent thymic emigrants to undergo LIP and convert into long-lived memory T cells. Surprisingly, however, most neonatal T cells do not undergo LIP. We therefore asked whether neonate-specific mechanisms prevent lymphopenia-driven T cell activation. In this study, we show that IL-7R-dependent innate lymphoid cells (ILCs) block LIP of CD8(+) T cells in neonatal but not adult mice. Importantly, CD8(+) T cell responses against a foreign Ag are not inhibited by neonatal ILCs. This ILC-based inhibition of LIP ensures the generation of a diverse naive T cell pool in lymphopenic neonates that is mandatory for the maintenance of T cell homeostasis and immunological self-tolerance later in life

IL-7R signaling in non-hematopoietic cells regulates CD8⁺ T cell differentiation in response to peptide vaccination and IL-7 therapy.

<p>(A-G) The indicated BM chimeras received CD8⁺ OT-I T cells and were treated with SIINFEKL and rIL-7 as described above. Three weeks after T cell transfer, splenic CD8⁺ OT-I T cells were analyzed by flow cytometry. Bar diagrams show pooled results (±SEM) of 2–3 independent experiments with a total of (A, B, C, E, G) 10–17 or (D) 4–10 mice/group. Histogram overlays show representative results for individual mice. Grey-lined histograms represent FMO staining controls.</p

The combination of rIL-7 therapy and peptide vaccination impairs T cell-dependent tumor rejection in Rag^-/- mice.

<p>(A) Rag^-/- and (B) Rag^-/-IL-7R^-/- mice were reconstituted with 1 x 10⁶ CD8⁺CD90.1⁺ OT-I T cells or were left untreated (+/- OT-I). One day later, OT-I-reconstituted mice were either vaccinated with 50 μg SIINFEKL or received PBS (+/- Pep). rIL-7 or PBS (+/- IL-7) were injected every 3–4 days for 19 days starting one day before T cell transfer. Mice were challenged s.c. with 1 x 10⁶ EG7 tumor cells three weeks after T cell transfer. Mice with tumors larger than 250 mm³ were scored as tumor positive. Shown are pooled data from 2 independent experiments with a total of 12–13 T cell reconstituted mice. Primary tumor growth was analyzed in untreated Rag^-/- and Rag^-/-IL-7R^-/- mice (n = 3). Statistical significance was calculated using the log-rank test. (C) The numbers of splenic DCs were determined in tumor-bearing mice 28–37 days after tumor challenge. Pooled data (±SEM) from 2 independent experiments with a total of 6–11 mice/group are shown. Statistical significance was calculated using the Mann-Whitney test.</p

Host IL-7R signaling is required for granulocyte and DC expansion in response to rIL-7.

<p>Rag^-/- and Rag^-/-IL-7R^-/- mice were treated with rIL-7 or PBS (+/- IL-7) every 3–4 days and spleens were analyzed by flow cytometry after 10–24 days. (A-E) Shown are numbers of (A) splenocytes (Splen.), (B) CD11b⁺Gr1⁺ granulocytes (Gran.), (C) CD11c⁺MHC-II⁺ dendritic cells (DCs), (D) CD8⁺ and (E) CD8^- DCs. Shown are pooled data (±SEM) from 2 independent experiments with a total of 7–8 mice/group.</p

Host IL-7R signaling modulates CD8⁺ T cell expansion and differentiation in response to peptide vaccination and IL-7 therapy.

<p>(A-J) Rag^-/- and Rag^-/-IL-7R^-/- mice were reconstituted with CD8⁺CD90.1⁺ OT-I T cells and treated with SIINFEKL +/- rIL-7 as described in Fig 4. Three weeks after T cell transfer (A, B) spleens and (C-J) peripheral blood were analyzed by flow cytometry. Shown are numbers of (A) splenocytes and (B) splenic CD8⁺CD90.1⁺ OT-I T cells. After gating on OT-I T cells, relative frequencies (frequ.) of (C) CD62L^hi, (E) KLRG-1^hi, (F) Ki67^hi, (H) IFN-γ^hi, (I) TNF-α^hi OT-I cells and relative MFIs for (D) CD127, (Γ) Bcl-2 and (J) PD-1 were determined. (H, I) Cytokine production was measured after <i>in vitro</i> stimulation of PBMCs with 1 μM SIINFEKL peptide in the presence of Rag^-/- splenocytes, brefeldin A and monensin for 6 hours. Relative (rel.) values were calculated as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0159690#pone.0159690.g002" target="_blank">Fig 2</a>. Bar diagrams show pooled data (±SEM) from 3 independent experiments with a total of 15–18 mice/group. Histogram overlays show representative results for individual mice. Dashed lines in overlays represent (C-G and J) FMO control samples or (H, I) stained cells without prior SIINFEKL stimulation.</p