Functional evolution of IGF2:IGF2R domain 11 binding generates novel structural interactions and a specific IGF2 antagonist

Among the 15 extracellular domains of the mannose 6-phosphate/insulin-like growth factor-2 receptor (M6P/IGF2R), domain 11 has evolved a binding site for IGF2 to negatively regulate ligand bioavailability and mammalian growth. Despite the highly evolved structural loops of the IGF2:domain 11 binding site, affinity-enhancing AB loop mutations suggest that binding is modifiable. Here we examine the extent to which IGF2:domain 11 affinity, and its specificity over IGF1, can be enhanced, and we examine the structural basis of the mechanistic and functional consequences. Domain 11 binding loop mutants were selected by yeast surface display combined with high-resolution structure-based predictions, and validated by surface plasmon resonance. We discovered previously unidentified mutations in the ligand-interacting surface binding loops (AB, CD, FG, and HI). Five combined mutations increased rigidity of the AB loop, as confirmed by NMR. When added to three independently identified CD and FG loop mutations that reduced the koff value by twofold, these mutations resulted in an overall selective 100-fold improvement in affinity. The structural basis of the evolved affinity was improved shape complementarity established by interloop (AB-CD) and intraloop (FG-FG) side chain interactions. The high affinity of the combinatorial domain 11 Fc fusion proteins functioned as ligand-soluble antagonists or traps that depleted pathological IGF2 isoforms from serum and abrogated IGF2-dependent signaling in vivo. An evolved and reengineered high-specificity M6P/IGF2R domain 11 binding site for IGF2 may improve therapeutic targeting of the frequent IGF2 gain of function observed in human cancer

The influence of confounders in the analysis of mid-regional pro-atrial natriuretic peptide in patients with chronic heart failure

Natriuretic peptides play an important role in the diagnosis and risk stratification of patients with acute and chronic heart failure. Multiple studies have shown that these peptides are liable to the influence of individual factors. For N-terminal-pro-B-type natriuretic peptide (NT-proBNP) some of these confounding factors have been evaluated over the years such as age, gender, New York Heart Association (NYHA) class and body mass index (BMI). The aim of this study was to establish confounding factors of mid-regional pro-atrial natriuretic peptide (MR-proANP) assessment

Mortality in Puerto Rico after Hurricane Maria

BACKGROUND Quantifying the effect of natural disasters on society is critical for recovery of public health services and infrastructure. The death toll can be difficult to assess in the aftermath of a major disaster. In September 2017, Hurricane Maria caused massive infrastructural damage to Puerto Rico, but its effect on mortality remains contentious. The official death count is 64. METHODS Using a representative, stratified sample, we surveyed 3299 randomly chosen households across Puerto Rico to produce an independent estimate of all-cause mortality after the hurricane. Respondents were asked about displacement, infrastructure loss, and causes of death. We calculated excess deaths by comparing our estimated post-hurricane mortality rate with official rates for the same period in 2016. RESULTS From the survey data, we estimated a mortality rate of 14.3 deaths (95% confidence interval [CI], 9.8 to 18.9) per 1000 persons from September 20 through December 31, 2017. This rate yielded a total of 4645 excess deaths during this period (95% CI, 793 to 8498), equivalent to a 62% increase in the mortality rate as compared with the same period in 2016. However, this number is likely to be an underestimate because of survivor bias. The mortality rate remained high through the end of December 2017, and one third of the deaths were attributed to delayed or interrupted health care. Hurricane-related migration was substantial. CONCLUSIONS This household-based survey suggests that the number of excess deaths related to Hurricane Maria in Puerto Rico is more than 70 times the official estimate. (Funded by the Harvard T.H. Chan School of Public Health and others.

Avian Influenza Viruses Infect Primary Human Bronchial Epithelial Cells Unconstrained by Sialic Acid α2,3 Residues

Avian influenza viruses (AIV) are an important emerging threat to public health. It is thought that sialic acid (sia) receptors are barriers in cross-species transmission where the binding preferences of AIV and human influenza viruses are sias α2,3 versus α2,6, respectively. In this study, we show that a normal fully differentiated, primary human bronchial epithelial cell model is readily infected by low pathogenic H5N1, H5N2 and H5N3 AIV, which primarily bind to sia α2,3 moieties, and replicate in these cells independent of specific sias on the cell surface. NHBE cells treated with neuraminidase prior to infection are infected by AIV despite removal of sia α2,3 moieties. Following AIV infection, higher levels of IP-10 and RANTES are secreted compared to human influenza virus infection, indicating differential chemokine expression patterns, a feature that may contribute to differences in disease pathogenesis between avian and human influenza virus infections in humans

Allergen Uptake, Activation, and IL-23 Production by Pulmonary Myeloid DCs Drives Airway Hyperresponsiveness in Asthma-Susceptible Mice

Maladaptive, Th2-polarized inflammatory responses are integral to the pathogenesis of allergic asthma. As regulators of T cell activation, dendritic cells (DCs) are important mediators of allergic asthma, yet the precise signals which render endogenous DCs “pro-asthmatic”, and the extent to which these signals are regulated by the pulmonary environment and host genetics, remains unclear. Comparative phenotypic and functional analysis of pulmonary DC populations in mice susceptible (A/J), or resistant (C3H) to experimental asthma, revealed that susceptibility to airway hyperresponsiveness is associated with preferential myeloid DC (mDC) allergen uptake, and production of Th17-skewing cytokines (IL-6, IL-23), whereas resistance is associated with increased allergen uptake by plasmacytoid DCs. Surprisingly, adoptive transfer of syngeneic HDM-pulsed bone marrow derived mDCs (BMDCs) to the lungs of C3H mice markedly enhanced lung IL-17A production, and rendered them susceptible to allergen-driven airway hyperresponsiveness. Characterization of these BMDCs revealed levels of antigen uptake, and Th17 promoting cytokine production similar to that observed in pulmonary mDCs from susceptible A/J mice. Collectively these data demonstrate that the lung environment present in asthma-resistant mice promotes robust pDC allergen uptake, activation, and limits Th17-skewing cytokine production responsible for driving pathologic T cell responses central to the development of allergen-induced airway hyperresponsiveness

31st Annual Meeting and Associated Programs of the Society for Immunotherapy of Cancer (SITC 2016) : part two

Background The immunological escape of tumors represents one of the main ob- stacles to the treatment of malignancies. The blockade of PD-1 or CTLA-4 receptors represented a milestone in the history of immunotherapy. However, immune checkpoint inhibitors seem to be effective in specific cohorts of patients. It has been proposed that their efficacy relies on the presence of an immunological response. Thus, we hypothesized that disruption of the PD-L1/PD-1 axis would synergize with our oncolytic vaccine platform PeptiCRAd. Methods We used murine B16OVA in vivo tumor models and flow cytometry analysis to investigate the immunological background. Results First, we found that high-burden B16OVA tumors were refractory to combination immunotherapy. However, with a more aggressive schedule, tumors with a lower burden were more susceptible to the combination of PeptiCRAd and PD-L1 blockade. The therapy signifi- cantly increased the median survival of mice (Fig. 7). Interestingly, the reduced growth of contralaterally injected B16F10 cells sug- gested the presence of a long lasting immunological memory also against non-targeted antigens. Concerning the functional state of tumor infiltrating lymphocytes (TILs), we found that all the immune therapies would enhance the percentage of activated (PD-1pos TIM- 3neg) T lymphocytes and reduce the amount of exhausted (PD-1pos TIM-3pos) cells compared to placebo. As expected, we found that PeptiCRAd monotherapy could increase the number of antigen spe- cific CD8+ T cells compared to other treatments. However, only the combination with PD-L1 blockade could significantly increase the ra- tio between activated and exhausted pentamer positive cells (p= 0.0058), suggesting that by disrupting the PD-1/PD-L1 axis we could decrease the amount of dysfunctional antigen specific T cells. We ob- served that the anatomical location deeply influenced the state of CD4+ and CD8+ T lymphocytes. In fact, TIM-3 expression was in- creased by 2 fold on TILs compared to splenic and lymphoid T cells. In the CD8+ compartment, the expression of PD-1 on the surface seemed to be restricted to the tumor micro-environment, while CD4 + T cells had a high expression of PD-1 also in lymphoid organs. Interestingly, we found that the levels of PD-1 were significantly higher on CD8+ T cells than on CD4+ T cells into the tumor micro- environment (p < 0.0001). Conclusions In conclusion, we demonstrated that the efficacy of immune check- point inhibitors might be strongly enhanced by their combination with cancer vaccines. PeptiCRAd was able to increase the number of antigen-specific T cells and PD-L1 blockade prevented their exhaus- tion, resulting in long-lasting immunological memory and increased median survival

Einleitung - zum Lehrbuch und dem etwas anderen Lehrbuchprojekt

L3T ist überarbeitet und steht nun in der zweiten Auflage zur Verfügung. Wie es dazu kam und wie es im Rahmen des Projekts "L3T 2.0" umgesetzt wurde, ist hier festgehalten. Da sich die Entstehung dieses Lehrbuchs deutlich von anderen Buchprojekten unterscheidet, wollen wir auch Einblick in den Entstehungsprozess geben. Ebenso, quasi als Vorwort in eigener Sache, wird beschrieben, worum es im Lehrbuch geht und wie man damit umgehen soll. So hat jedes Kapitel ein eigenes Schlagwort, einen Hashtag (#), auf den sich einerseits die Kapitel untereinander beziehen und mit denen andererseits im World Wide Web weitere Materialien bei verschiedenen Serviceangeboten zugänglich sind. Schließlich sprechen wir in dieser Einleitung noch jede Menge Danksagungen aus, denn das Buch ist nicht das Werk von wenigen Personen, sondern ein Projekt, bei dem immerhin rund 250 Personen mit- und zusammengewirkt haben! (DIPF/Orig.

Universit\ue4ten in sozialen Netzwerken. Wie Hochschulen die Chancen und Herausforderungen der sozialen Medien nutzen k\uf6nnen

Soziale Medien haben l\ue4ngst die Grenze zur Allgegenw\ue4rtigkeit erreicht und sind heutzutage nicht nur in unserem Privatleben allt\ue4glicher Begleiter, sondern werden auch von Unternehmen, Organisationen und anderen nicht reellen Personen f\ufcr die unterschiedlichsten Zwecke eingesetzt. Zunehmend versuchen auch Bildungs- und Ausbildungseinrichtungen diese Medien f\ufcr sich zu erschlie fen. Neue Handlungsfelder und vor allem auch neue Wege zur Selbstdarstellung werden dabei erprobt und auf einen potenziellen Mehrwert untersucht. Kernpunkt ist die Frage, wie Social-Media-Aktivit\ue4ten aussehen sollen, um als erfolgreich und wertvoll angesehen zu werden, welche Einflussfaktoren sich auf die Wahrnehmung der Nutzer auswirken und welche internen Mechanismen den sozialen Medien zu Grunde liegen und so in weiterem Sinne auch deren Inhalte bedingen. Um diesen Fragen nachzugehen, wurden f\ufcr den Bereich der 6ffentlichkeitsarbeit mittels sozialer Netzwerke an Hochschulen Untersuchungen angestellt, die die Wirksamkeit unterschiedlicher Herangehensweisen analysierten und versuchten, daraus Handlungsfelder f\ufcr zuk\ufcnftige Social-Media-Aktivit\ue4ten an Universit\ue4ten und Fachhochschulen zu identifizieren. Dabei stellte sich heraus, dass die Beteiligung der Nutzer durchaus mit unterschiedlichen Eigenschaften einzelner Social-Media-Aktivit\ue4ten zusammenh\ue4ngt und dass zum Beispiel der Ver\uf6ffentlichungszeitpunkt, die verwendeten Elemente eines Posts und vor allem der Inhalt besonders hervortreten. Die Anzahl der Postings pro Woche und die Textl\ue4nge sind jedoch weniger relevant. Allgemein l\ue4sst sich sagen, dass un\ufcbliche Posting-Zeitpunkte, eine Kombination aus Bild, Text und Hyperlink sowie soziale, teilweise emotionale, teilweise auch stark originalit\ue4tsentbundene, konventionalisierte Inhalte, die die Gemeinschaft hervorheben und eine Kontaktpflege innerhalb ihrer erm\uf6glichen, die meistgelesenen und involvierendsten Beitr\ue4ge in sozialen Netzwerken darstellen

Das Projekt L3T 2.0 im cberblick \u2013 Organisation und Abl\ue4ufe

In diesem Beitrag wird das Projekt L3T 2.0, der Ablauf des gesamten Projekts sowie die Planungen f\ufcr die sieben Tage vorgestellt. Dazu werden Aufgaben, die vergeben wurden, gelistet, Abl\ue4ufe skizziert sowie eingesetzte Werkzeuge beschrieben