An Expanded Polyproline Domain Maintains Mutant Huntingtin Soluble in vivo and During Aging.

Funder: Alzheimer’s Research UKFunder: Michael J. Fox Foundation for Parkinson’s ResearchFunder: Medical Research CouncilFunder: Deutsche ForschungsgemeinschaftHuntington's disease is a dominantly inherited neurodegenerative disorder caused by the expansion of a CAG repeat, encoding for the amino acid glutamine (Q), present in the first exon of the protein huntingtin. Over the threshold of Q39 HTT exon 1 (HTTEx1) tends to misfold and aggregate into large intracellular structures, but whether these end-stage aggregates or their on-pathway intermediates are responsible for cytotoxicity is still debated. HTTEx1 can be separated into three domains: an N-terminal 17 amino acid region, the polyglutamine (polyQ) expansion and a C-terminal proline rich domain (PRD). Alongside the expanded polyQ, these flanking domains influence the aggregation propensity of HTTEx1: with the N17 initiating and promoting aggregation, and the PRD modulating it. In this study we focus on the first 11 amino acids of the PRD, a stretch of pure prolines, which are an evolutionary recent addition to the expanding polyQ region. We hypothesize that this proline region is expanding alongside the polyQ to counteract its ability to misfold and cause toxicity, and that expanding this proline region would be overall beneficial. We generated HTTEx1 mutants lacking both flanking domains singularly, missing the first 11 prolines of the PRD, or with this stretch of prolines expanded. We then followed their aggregation landscape in vitro with a battery of biochemical assays, and in vivo in novel models of C. elegans expressing the HTTEx1 mutants pan-neuronally. Employing fluorescence lifetime imaging we could observe the aggregation propensity of all HTTEx1 mutants during aging and correlate this with toxicity via various phenotypic assays. We found that the presence of an expanded proline stretch is beneficial in maintaining HTTEx1 soluble over time, regardless of polyQ length. However, the expanded prolines were only advantageous in promoting the survival and fitness of an organism carrying a pathogenic stretch of Q48 but were extremely deleterious to the nematode expressing a physiological stretch of Q23. Our results reveal the unique importance of the prolines which have and still are evolving alongside expanding glutamines to promote the function of HTTEx1 and avoid pathology

Modulation of immune responses using adjuvants to facilitate therapeutic vaccination

Publsher's version (útgefin grein)Therapeutic vaccination offers great promise as an intervention for a diversity of infectious and non-infectious conditions. Given that most chronic health conditions are thought to have an immune component, vaccination can at least in principle be proposed as a therapeutic strategy. Understanding the nature of protective immunity is of vital importance, and the progress made in recent years in defining the nature of pathological and protective immunity for a range of diseases has provided an impetus to devise strategies to promote such responses in a targeted manner. However, in many cases, limited progress has been made in clinical adoption of such approaches. This in part results from a lack of safe and effective vaccine adjuvants that can be used to promote protective immunity and/or reduce deleterious immune responses. Although somewhat simplistic, it is possible to divide therapeutic vaccine approaches into those targeting conditions where antibody responses can mediate protection and those where the principal focus is the promotion of effector and memory cellular immunity or the reduction of damaging cellular immune responses as in the case of autoimmune diseases. Clearly, in all cases of antigen-specific immunotherapy, the identification of protective antigens is a vital first step. There are many challenges to developing therapeutic vaccines beyond those associated with prophylactic diseases including the ongoing immune responses in patients, patient heterogeneity, and diversity in the type and stage of disease. If reproducible biomarkers can be defined, these could allow earlier diagnosis and intervention and likely increase therapeutic vaccine efficacy. Current immunomodulatory approaches related to adoptive cell transfers or passive antibody therapy are showing great promise, but these are outside the scope of this review which will focus on the potential for adjuvanted therapeutic active vaccination strategies.This article/publication is based upon work from COST Action CA16231 ENOVA (European Network of Vaccine Adjuvants), supported by COST (European Cooperation in Science and Technology—www.cost.eu).Peer Reviewe

Induction of potent neutralizing antibody responses by a designed protein nanoparticle accine for respiratory syncytial virus

Respiratory syncytial virus (RSV) is a worldwide public health concern for which no vaccine is available. Elucidation of the prefusion structure of the RSV F glycoprotein and its identification as the main target of neutralizing antibodies have provided new opportunities for development of an effective vaccine. Here, we describe the structure-based design of a self-assembling protein nanoparticle presenting a prefusion-stabilized variant of the F glycoprotein trimer (DS-Cav1) in a repetitive array on the nanoparticle exterior. The two-component nature of the nanoparticle scaffold enabled the production of highly ordered, monodisperse immunogens that display DS-Cav1 at controllable density. In mice and nonhuman primates, the full-valency nanoparticle immunogen displaying 20 DS-Cav1 trimers induced neutralizing antibody responses ∼10-fold higher than trimeric DS-Cav1. These results motivate continued development of this promising nanoparticle RSV vaccine candidate and establish computationally designed two-component nanoparticles as a robust and customizable platform for structure-based vaccine design

Alternativen zu Säuglingsnahrungen auf Kuhmilchproteinbasis

Die natürliche Ernährung eines gesunden Säuglings ist das Stillen. Ist Stillen oder die Fütterung von Muttermilch nicht möglich, kann Frauenmilch für die Ernährung des Säuglings erwogen werden, sofern diese aus einer qualifizierten Humanmilchbank stammt. Vom Kauf von Frauenmilch aus dem Internet wird wegen Risiken einer Infektionsübertragung und einer unzureichenden Milchqualität strikt abgeraten. Aufgrund der geringen Verfügbarkeit, der hohen Kosten sowie möglicher Nachteile der Ernährung mit Spendermilch im Vergleich zum Stillen sind industriell gefertigte Säuglingsnahrungen das Mittel der Wahl zur Ernährung des gesunden, reifgeborenen Säuglings, wenn Stillen nicht oder nur partiell möglich ist. Kuhmilchprotein ist die am häufigsten verwendete Eiweißkomponente. Die Nachfrage nach anderen auf Tiermilchen basierten sowie vegetarischen bzw. veganen Alternativen steigt. Im Folgenden werden verschiedene Alternativen bezüglich ihrer Eignung betrachtet. Säuglingsnahrungen auf Basis von Ziegenmilchprotein stellen für gesunde, reifgeborene Säuglinge eine zugelassene und geeignete Alternative zu kuhmilchproteinbasierten Säuglingsnahrungen dar. Für Säuglingsnahrungen aus anderen Tiermilchen (z. B. Kamel‑, Schaf‑, Pferde- oder Büffelmilch) sind keine belastbaren Daten zu Eignung und Sicherheit bekannt, und sie sind in der Europäischen Union nicht zugelassen. Säuglingsnahrungen auf der Grundlage von Sojaproteinisolaten sind in der Europäischen Union zugelassen. Sie werden für die allgemeine Verwendung im 1. Lebenshalbjahr durch die Ernährungskommission nicht empfohlen, insbesondere weil potenziell nachteilige Effekte von enthaltenen Isoflavonen nicht ausgeschlossen werden können. Ab der Geburt und in den ersten Lebensmonaten sollte die Gabe von Sojanahrungen auf Indikationen wie eine bestehende Galaktosämie, die sehr seltene kongenitale Laktoseintoleranz sowie bei familiärem Wunsch nach veganer Ernährungsweise und aus anderen weltanschaulichen Gründen begrenzt werden. Im 2. Lebenshalbjahr ist die Zufuhrmenge pro Kilogramm Körpergewicht deutlich niedriger, sodass das Risiko unerwünschter Wirkungen als wesentlich geringer eingeschätzt wird. Zu neuerdings angebotenen Nahrungen auf der Grundlage einer Mischung aus Soja- und Kuhmilchprotein sind keine Daten zur Prüfung von Sicherheit und Eignung bekannt, sodass hierzu keine Empfehlung ausgesprochen werden kann. Von der Verwendung von Säuglingsnahrung auf der Grundlage von hydrolysiertem Reisprotein wird auch aufgrund hoher berichteter Arsengehalte abgeraten. Auch von einer häuslichen Selbstherstellung von Säuglingsnahrungen wird aufgrund eines erhöhten Risikos für eine nichtbedarfsgerechte Nährstoffzufuhr und für Infektionen abgeraten. // The natural nutrition of a healthy infant is breastfeeding. If breastfeeding or feeding of breast milk is not possible, feeding of donor human milk may be considered. The safety of donor milk can only be ensured if it is provided by a qualified human milk bank. The use of informally shared donor milk or the use of human milk purchased through the internet is strongly discouraged because of the risk of transmitting infections and of insufficient milk quality. Due to low availability, high costs and concerns about poorer nutritional quality of donor milk compared to breastfeeding, industrially manufactured infant formula is the preferred alternative for feeding healthy full-term infants that cannot or cannot be fully breastfed. Milk-based infant formula is most frequently made from cow’s milk protein; however, there is an increasing popularity of vegetarian or vegan formulas and of formulas based on different animal milks other than cow’s milk. Goat’s milk-based infant formulas represent an approved and suitable alternative to cow’s milk for healthy, full-term infants. Reliable data on the suitability and safety are unavailable for formulas based on other animal milks (e.g., camel, horse, sheep or buffalo milk), and these are not approved for infant feeding in the European Union. Infant formulas based on soybean protein isolates are approved in the European Union. They are not generally recommended by the Nutrition Committee for infant feeding in the first half year of life because potentially harmful effects of isoflavones derived from soybeans cannot be excluded; however, soybean protein isolate-based formulas may be reservedly used after birth and in the first months of life for infants affected by galactosemia, the very rare hereditary lactose intolerance manifest at birth, a vegan family lifestyle or other familial convictions. In the second half year of life the intake amount per kilogram body weight is much less so that the risk of undesired effects is estimated to be much lower. For the recently provided nutrition based on a mixture of soybean and cow’s milk proteins, no data on testing of the safety and suitability are known, so that no recommendations can be made on this. The use of infant formula based on hydrolyzed rice protein is not recommended because of concerns about possible high arsenic contamination. The use of homemade infant formulas is discouraged due to the high risk of introducing infections and of possible nutritional imbalances of macronutrients and micronutrients

Der TPL-2 - ERK Signalweg in chronisch entzündlichen Darmerkrankungen

Chronisch-entzündliche Darmerkrankungen wie Morbus Crohn und Colitis ulcerosa sind als unkontrollierte Immunreaktionen gegenüber der Darmflora beschrieben, die zur chronischen Entzündung des Darmes führen. In der Darmschleimhaut von Morbus Crohn Patienten wurde eine verstärkte NF-ĸB Aktivität nachgewiesen (Visekruna et al., 2006). In diesem Zusammenhang wurden auch erhöhte Mengen von pro-inflammatorische Zytokinen TNFα und IL-1β beschrieben (Schreiber et al., 1999), die ausschlaggebend an der Entzündung im Darm beteiligt sind. Neben NF-ĸB wird die Produktion von TNFα und IL-1β auch von dem MAP Kinase Signalweg TPL-2-ERK reguliert. Wir konnten zeigen, dass der TPL-2-ERK Signalweg im entzündeten Darmgewebe von Morbus Crohn Patienten aber nicht in Patienten mit Colitis ulcerosa aktiviert ist. Weiterhin konnten wir im Mausmodell zeigen, dass das Fehlen von TPL-2-ERK Signalen zu einer Verringerung der Darmentzündung führt. Dieser Effekt wurde vor allem durch die Unterdrückung der pro-inflammatorischen Zytokine TNFα, IL-1β und IL-6 herbeigeführt, dessen Produktion von bakteriellen Bestandteilen stimuliert wird. Unsere Ergebnisse zeigen deutlich, dass neben NF-ĸB Signalen die Aktivierung des TPL-2-ERK Signalweges zu einer verstärkten Produktion von pro-inflammatorischen Zytokinen und damit zur Darmentzündung führt. Weiterhin konnten wir zeigen, dass die Unterdrückung der Zytokinproduktion bei fehlenden TPL-2 Signale zum Teil durch die Anhäufung von inaktiven NF-ĸB p50 Homodimeren im Zellkern entsteht. LPS-Toleranz führt ebenfalls zu einer Akkumulation von NF-ĸB Homodimeren (Ziegler-Heitbrock et al., 1994) , die eine LPS-induzierte Transkription effektiv blockieren. Mit der Hemmung von TPL-2 Signalen zeigen wir also einen neuen Mechanismus mit dem NF-ĸB Aktivierung nach Immunstimulation moduliert werden kann und ohne die NF-ĸB vermittelten protektiven Zellfunktionen zu stören. Diese Eigenschaft macht die TPL-2 Kinase zu einem interessanten Zielprotein für entzündungshemmende Medikamente, die potentiell für die Behandlung von Morbus Crohn Patienten eingesetzt werden könnten.Inflammatory bowel disease, namely Crohn’s disease and Ulcerative colitis, are characterized by dysregulated immune responses to the intestinal microbiota leading to chronic inflammation of the colon and small intestine. Prolonged and enhanced activation of NF-ĸB was shown in the intestinal lamina propria of Crohn's disease patients (Visekruna et al., 2006). This correlates with overproduction of the proinflammatory cytokines TNFα and IL-1β (Schreiber et al., 1999), which are among the most important cytokines that drive intestinal inflammation. In addition to NF-ĸB, the MAP kinase pathway TPL-2-ERK also regulates synthesis of TNFα and IL-1β. We found that the TPL-2-ERK pathway is activated in the inflamed intestinal tissue of patients with Crohn’s disease but not in Ulcerative colitis patients. Furthermore, by studying the murine DSS-colitis model we found that the absence of TPL-2-ERK signaling ameliorates intestinal inflammation. This was mainly due to inhibited production of the pro-inflammatory cytokines TNFα, IL-1β and IL-6 in response to bacterial components. Therefore, our results clearly show that in addition to NF-ĸB signaling increased TPL-2-ERK activation is responsible for enhanced production of pro-inflammatory cytokines leading to intestinal inflammation. Interestingly, we found that cytokine suppression in the absence of TPL-2 signals was at least partly mediated by accumulation of transcriptional inactive NF-ĸB p50 homodimers in the nucleus. NF-ĸB homodimer accumulation also occurs during LPS tolerance and displays an effective mechanism to block LPS induced transcription (Ziegler-Heitbrock et al., 1994). Thus, by targeting TPL-2, we found a new mechanism to modulate NF-ĸB activation after immune stimulation, while leaving NF-ĸB mediated protective cell functions undisturbed. These properties of TPL-2 kinase make it a promising new target for an anti-inflammatory drug that may have potential use in the treatment of Crohn’s disease patients

Illness Perception, Knowledge and Self-Care about Cervical Cancer

Prevention plays a central role in early detection of cervical cancer. Common Sense Model proposes that the nature and organization of illness representations can guide actions related to health and how self-care is exercised. The aim of this study was to describe and compare illness perception, knowledge and self-care in women with and without cancer precursor lesions. Participants were 92 women (aged 18-59) from primary care unity divided into two groups: women with and without premalignant lesion. Measures for illness perception, knowledge and self-care were used. There was no statistically significant difference (t test e chi-square test) between groups in the variables analyzed. Despite the risk for cervical cancer, women with precursor lesions do not adjust their illness perceptions, knowledge and self-care to the situation. These data show the need to warn women against the cervical cancer risks, because their distorted perceptions and lack of knowledge about the disease may hamper the screening and control of cervical cancer

Discovering Yersinia–Host Interactions by Tissue Dual RNA-Seq

Kusmierek M, Heroven AK, Beckstette M, Nuss AM, Dersch P. Discovering Yersinia–Host Interactions by Tissue Dual RNA-Seq. In: Vadyvaloo V, Lawrenz MB, eds. Pathogenic Yersinia. Methods and Protocols. Methods in Molecular Biology. Vol 2010. New York, NY: Springer ; 2019: 99-116

Vaccine-induced Th1-type response protects against invasive group A Streptococcus infection in the absence of opsonizing antibodies

Recent global advocacy efforts have highlighted the importance of development of a vaccine against group A Streptococcus (GAS). Combo5 is a non-M protein-based vaccine that provides protection against GAS skin infection in mice and reduces the severity of pharyngitis in nonhuman primates. However, Combo5 with the addition of aluminum hydroxide (alum) as an adjuvant failed to protect against invasive GAS infection of mice. Here, we show that formulation of Combo5 with adjuvants containing saponin QS21 significantly improves protective efficacy, even though all 7 adjuvants tested generated high antigen-specific IgG antibody titers, including alum. Detailed characterization of Combo5 formulated with SMQ adjuvant, a squalene-in-water emulsion containing a TLR4 agonist and QS21, showed significant differences from the results obtained with alum in IgG subclasses generated following immunization, with an absence of GAS opsonizing antibodies. SMQ, but not alum, generated strong interleukin-6 (IL-6), gamma interferon (IFN-γ), and tumor necrosis alpha (TNF-α) responses. This work highlights the importance of adjuvant selection for non-M protein-based GAS vaccines to optimize immune responses and protective efficacy.IMPORTANCE Availability of a group A Streptococcus vaccine remains an unmet public health need. Here, we tested different adjuvant formulations to improve the protective efficacy of non-M protein vaccine Combo5 in an invasive disease model. We show that novel adjuvants can dramatically shape the type of immune response developed following immunization with Combo5 and significantly improve protection. In addition, protection afforded by Combo5 is not mediated by opsonizing antibodies, believed to be the main correlate of protection against GAS infections. Overall, this report highlights the importance of adjuvant selection in raising protective immune responses against GAS invasive infection. Adjuvants that can provide a more balanced Th1/Th2-type response may be required to optimize protection of GAS vaccines, particularly those based on non-M protein antigens