Pituitary Adenylate Cyclase-Activating Polypeptide—A Neuropeptide as Novel Treatment Option for Subacute Ileitis in Mice Harboring a Human Gut Microbiota

The neuropeptide Pituitary adenylate cyclase-activating polypeptide (PACAP) is well-known for its important functions in immunity and inflammation. Data regarding anti-inflammatory properties of PACAP in the intestinal tract are limited, however. In our present preclinical intervention study we addressed whether PACAP treatment could alleviate experimental subacute ileitis mimicking human gut microbiota conditions. Therefore, secondary abioitic mice were subjected to human fecal microbiota transplantation (FMT) and perorally infected with low-dose Toxoplasma gondii to induce subacute ileitis on day 0. From day 3 until day 8 post-infection, mice were either treated with synthetic PACAP38 or placebo. At day 9 post-infection, placebo, but not PACAP treated mice exhibited overt macroscopic sequelae of intestinal immunopathology. PACAP treatment further resulted in less distinct apoptotic responses in ileal and colonic epithelia that were accompanied by lower T cell numbers in the mucosa and lamina propria and less secretion of pro-inflammatory cytokines in intestinal ex vivo biopsies. Notably, ileitis-associated gut microbiota shifts were less distinct in PACAP as compared to placebo treated mice. Inflammation-ameliorating effects of PACAP were not restricted to the intestines, but could also be observed in extra-intestinal including systemic compartments as indicated by lower apoptotic cell counts and less pro-inflammatory cytokine secretion in liver and lungs taken from PACAP treated as compared to placebo control mice, which also held true for markedly lower serum TNF and IL-6 concentrations in the former as compared to the latter. Our preclinical intervention study provides strong evidence that synthetic PACAP alleviates subacute ileitis and extra-intestinal including systemic sequelae of T cell-driven immunopathology. These findings further support PACAP as a novel treatment option for intestinal inflammation including inflammatory bowel diseases (IBD)

Pituitary Adenylate Cyclase-Activating Polypeptide Ameliorates Experimental Acute Ileitis and Extra-Intestinal Sequelae

Background The neuropeptide Pituitary adenylate cyclase-activating polypeptide (PACAP) plays pivotal roles in immunity and inflammation. So far, potential immune-modulatory properties of PACAP have not been investigated in experimental ileitis. Methodology/Principal Findings Mice were perorally infected with Toxoplasma (T.) gondii to induce acute ileitis (day 0) and treated daily with synthetic PACAP38 from day 1 to 6 post infection (p.i.; prophylaxis) or from day 4 to 6 p.i. (therapy). Whereas placebo-treated control mice suffered from acute ileitis at day 7 p.i. and succumbed to infection, intestinal immunopathology was ameliorated following PACAP prophylaxis. PACAP-treated mice exhibited increased abundance of small intestinal FOXP3+ cells, but lower numbers of ileal T lymphocytes, neutrophils, monocytes and macrophages, which was accompanied by less ileal expression of pro-inflammatory cytokines such as IL-23p19, IL-22, IFN-γ, and MCP-1. Furthermore, PACAP-treated mice displayed higher anti-inflammatory IL-4 concentrations in mesenteric lymph nodes and liver and higher systemic anti- inflammatory IL-10 levels in spleen and serum as compared to control animals at day 7 p.i. Remarkably, PACAP-mediated anti-inflammatory effects could also be observed in extra-intestinal compartments as indicated by reduced pro- inflammatory mediator levels in spleen (TNF-α, nitric oxide) and liver (TNF-α, IFN-γ, MCP-1, IL-6) and less severe histopathological sequelae in lungs and kidneys following prophylactic PACAP treatment. Strikingly, PACAP prolonged survival of T. gondii infected mice in a time-of-treatment dependent manner. Conclusion/Significance Synthetic PACAP ameliorates acute small intestinal inflammation and extra-intestinal sequelae by down-regulating Th1-type immunopathology, reducing oxidative stress and up-regulating anti-inflammatory cytokine responses. These findings provide novel potential treatment options of inflammatory bowel diseases

Selective targeting of pro-inflammatory Th1 cells by microRNA-148a-specific antagomirs in vivo.

In T lymphocytes, expression of miR-148a is induced by T-bet and Twist1, and is specific for pro-inflammatory Th1 cells. In these cells, miR-148a inhibits the expression of the pro-apoptotic protein Bim and promotes their survival. Here we use sequence-specific cholesterol-modified oligonucleotides against miR-148a (antagomir-148a) for the selective elimination of pro-inflammatory Th1 cells in vivo. In the murine model of transfer colitis, antagomir-148a treatment reduced the number of pro-inflammatory Th1 cells in the colon of colitic mice by 50% and inhibited miR-148a expression by 71% in the remaining Th1 cells. Expression of Bim protein in colonic Th1 cells was increased. Antagomir-148a-mediated reduction of Th1 cells resulted in a significant amelioration of colitis. The effect of antagomir-148a was selective for chronic inflammation. Antigen-specific memory Th cells that were generated by an acute immune reaction to nitrophenylacetyl-coupled chicken gamma globulin (NP-CGG) were not affected by treatment with antagomir-148a, both during the effector and the memory phase. In addition, antibody titers to NP-CGG were not altered. Thus, antagomir-148a might qualify as an effective drug to selectively deplete pro-inflammatory Th1 cells of chronic inflammation without affecting the protective immunological memory

Human Practice. Digital Ecologies. Our Future. : 14. Internationale Tagung Wirtschaftsinformatik (WI 2019) : Tagungsband

Erschienen bei: universi - Universitätsverlag Siegen. - ISBN: 978-3-96182-063-4Aus dem Inhalt: Track 1: Produktion & Cyber-Physische Systeme Requirements and a Meta Model for Exchanging Additive Manufacturing Capacities Service Systems, Smart Service Systems and Cyber- Physical Systems—What’s the difference? Towards a Unified Terminology Developing an Industrial IoT Platform – Trade-off between Horizontal and Vertical Approaches Machine Learning und Complex Event Processing: Effiziente Echtzeitauswertung am Beispiel Smart Factory Sensor retrofit for a coffee machine as condition monitoring and predictive maintenance use case Stakeholder-Analyse zum Einsatz IIoT-basierter Frischeinformationen in der Lebensmittelindustrie Towards a Framework for Predictive Maintenance Strategies in Mechanical Engineering - A Method-Oriented Literature Analysis Development of a matching platform for the requirement-oriented selection of cyber physical systems for SMEs Track 2: Logistic Analytics An Empirical Study of Customers’ Behavioral Intention to Use Ridepooling Services – An Extension of the Technology Acceptance Model Modeling Delay Propagation and Transmission in Railway Networks What is the impact of company specific adjustments on the acceptance and diffusion of logistic standards? Robust Route Planning in Intermodal Urban Traffic Track 3: Unternehmensmodellierung & Informationssystemgestaltung (Enterprise Modelling & Information Systems Design) Work System Modeling Method with Different Levels of Specificity and Rigor for Different Stakeholder Purposes Resolving Inconsistencies in Declarative Process Models based on Culpability Measurement Strategic Analysis in the Realm of Enterprise Modeling – On the Example of Blockchain-Based Initiatives for the Electricity Sector Zwischenbetriebliche Integration in der Möbelbranche: Konfigurationen und Einflussfaktoren Novices’ Quality Perceptions and the Acceptance of Process Modeling Grammars Entwicklung einer Definition für Social Business Objects (SBO) zur Modellierung von Unternehmensinformationen Designing a Reference Model for Digital Product Configurators Terminology for Evolving Design Artifacts Business Role-Object Specification: A Language for Behavior-aware Structural Modeling of Business Objects Generating Smart Glasses-based Information Systems with BPMN4SGA: A BPMN Extension for Smart Glasses Applications Using Blockchain in Peer-to-Peer Carsharing to Build Trust in the Sharing Economy Testing in Big Data: An Architecture Pattern for a Development Environment for Innovative, Integrated and Robust Applications Track 4: Lern- und Wissensmanagement (e-Learning and Knowledge Management) eGovernment Competences revisited – A Literature Review on necessary Competences in a Digitalized Public Sector Say Hello to Your New Automated Tutor – A Structured Literature Review on Pedagogical Conversational Agents Teaching the Digital Transformation of Business Processes: Design of a Simulation Game for Information Systems Education Conceptualizing Immersion for Individual Learning in Virtual Reality Designing a Flipped Classroom Course – a Process Model The Influence of Risk-Taking on Knowledge Exchange and Combination Gamified Feedback durch Avatare im Mobile Learning Alexa, Can You Help Me Solve That Problem? - Understanding the Value of Smart Personal Assistants as Tutors for Complex Problem Tasks Track 5: Data Science & Business Analytics Matching with Bundle Preferences: Tradeoff between Fairness and Truthfulness Applied image recognition: guidelines for using deep learning models in practice Yield Prognosis for the Agrarian Management of Vineyards using Deep Learning for Object Counting Reading Between the Lines of Qualitative Data – How to Detect Hidden Structure Based on Codes Online Auctions with Dual-Threshold Algorithms: An Experimental Study and Practical Evaluation Design Features of Non-Financial Reward Programs for Online Reviews: Evaluation based on Google Maps Data Topic Embeddings – A New Approach to Classify Very Short Documents Based on Predefined Topics Leveraging Unstructured Image Data for Product Quality Improvement Decision Support for Real Estate Investors: Improving Real Estate Valuation with 3D City Models and Points of Interest Knowledge Discovery from CVs: A Topic Modeling Procedure Online Product Descriptions – Boost for your Sales? Entscheidungsunterstützung durch historienbasierte Dienstreihenfolgeplanung mit Pattern A Semi-Automated Approach for Generating Online Review Templates Machine Learning goes Measure Management: Leveraging Anomaly Detection and Parts Search to Improve Product-Cost Optimization Bedeutung von Predictive Analytics für den theoretischen Erkenntnisgewinn in der IS-Forschung Track 6: Digitale Transformation und Dienstleistungen Heuristic Theorizing in Software Development: Deriving Design Principles for Smart Glasses-based Systems Mirroring E-service for Brick and Mortar Retail: An Assessment and Survey Taxonomy of Digital Platforms: A Platform Architecture Perspective Value of Star Players in the Digital Age Local Shopping Platforms – Harnessing Locational Advantages for the Digital Transformation of Local Retail Outlets: A Content Analysis A Socio-Technical Approach to Manage Analytics-as-a-Service – Results of an Action Design Research Project Characterizing Approaches to Digital Transformation: Development of a Taxonomy of Digital Units Expectations vs. Reality – Benefits of Smart Services in the Field of Tension between Industry and Science Innovation Networks and Digital Innovation: How Organizations Use Innovation Networks in a Digitized Environment Characterising Social Reading Platforms— A Taxonomy-Based Approach to Structure the Field Less Complex than Expected – What Really Drives IT Consulting Value Modularity Canvas – A Framework for Visualizing Potentials of Service Modularity Towards a Conceptualization of Capabilities for Innovating Business Models in the Industrial Internet of Things A Taxonomy of Barriers to Digital Transformation Ambidexterity in Service Innovation Research: A Systematic Literature Review Design and success factors of an online solution for cross-pillar pension information Track 7: IT-Management und -Strategie A Frugal Support Structure for New Software Implementations in SMEs How to Structure a Company-wide Adoption of Big Data Analytics The Changing Roles of Innovation Actors and Organizational Antecedents in the Digital Age Bewertung des Kundennutzens von Chatbots für den Einsatz im Servicedesk Understanding the Benefits of Agile Software Development in Regulated Environments Are Employees Following the Rules? On the Effectiveness of IT Consumerization Policies Agile and Attached: The Impact of Agile Practices on Agile Team Members’ Affective Organisational Commitment The Complexity Trap – Limits of IT Flexibility for Supporting Organizational Agility in Decentralized Organizations Platform Openness: A Systematic Literature Review and Avenues for Future Research Competence, Fashion and the Case of Blockchain The Digital Platform Otto.de: A Case Study of Growth, Complexity, and Generativity Track 8: eHealth & alternde Gesellschaft Security and Privacy of Personal Health Records in Cloud Computing Environments – An Experimental Exploration of the Impact of Storage Solutions and Data Breaches Patientenintegration durch Pfadsysteme Digitalisierung in der Stressprävention – eine qualitative Interviewstudie zu Nutzenpotenzialen User Dynamics in Mental Health Forums – A Sentiment Analysis Perspective Intent and the Use of Wearables in the Workplace – A Model Development Understanding Patient Pathways in the Context of Integrated Health Care Services - Implications from a Scoping Review Understanding the Habitual Use of Wearable Activity Trackers On the Fit in Fitness Apps: Studying the Interaction of Motivational Affordances and Users’ Goal Orientations in Affecting the Benefits Gained Gamification in Health Behavior Change Support Systems - A Synthesis of Unintended Side Effects Investigating the Influence of Information Incongruity on Trust-Relations within Trilateral Healthcare Settings Track 9: Krisen- und Kontinuitätsmanagement Potentiale von IKT beim Ausfall kritischer Infrastrukturen: Erwartungen, Informationsgewinnung und Mediennutzung der Zivilbevölkerung in Deutschland Fake News Perception in Germany: A Representative Study of People’s Attitudes and Approaches to Counteract Disinformation Analyzing the Potential of Graphical Building Information for Fire Emergency Responses: Findings from a Controlled Experiment Track 10: Human-Computer Interaction Towards a Taxonomy of Platforms for Conversational Agent Design Measuring Service Encounter Satisfaction with Customer Service Chatbots using Sentiment Analysis Self-Tracking and Gamification: Analyzing the Interplay of Motivations, Usage and Motivation Fulfillment Erfolgsfaktoren von Augmented-Reality-Applikationen: Analyse von Nutzerrezensionen mit dem Review-Mining-Verfahren Designing Dynamic Decision Support for Electronic Requirements Negotiations Who is Stressed by Using ICTs? A Qualitative Comparison Analysis with the Big Five Personality Traits to Understand Technostress Walking the Middle Path: How Medium Trade-Off Exposure Leads to Higher Consumer Satisfaction in Recommender Agents Theory-Based Affordances of Utilitarian, Hedonic and Dual-Purposed Technologies: A Literature Review Eliciting Customer Preferences for Shopping Companion Apps: A Service Quality Approach The Role of Early User Participation in Discovering Software – A Case Study from the Context of Smart Glasses The Fluidity of the Self-Concept as a Framework to Explain the Motivation to Play Video Games Heart over Heels? An Empirical Analysis of the Relationship between Emotions and Review Helpfulness for Experience and Credence Goods Track 11: Information Security and Information Privacy Unfolding Concerns about Augmented Reality Technologies: A Qualitative Analysis of User Perceptions To (Psychologically) Own Data is to Protect Data: How Psychological Ownership Determines Protective Behavior in a Work and Private Context Understanding Data Protection Regulations from a Data Management Perspective: A Capability-Based Approach to EU-GDPR On the Difficulties of Incentivizing Online Privacy through Transparency: A Qualitative Survey of the German Health Insurance Market What is Your Selfie Worth? A Field Study on Individuals’ Valuation of Personal Data Justification of Mass Surveillance: A Quantitative Study An Exploratory Study of Risk Perception for Data Disclosure to a Network of Firms Track 12: Umweltinformatik und nachhaltiges Wirtschaften Kommunikationsfäden im Nadelöhr – Fachliche Prozessmodellierung der Nachhaltigkeitskommunikation am Kapitalmarkt Potentiale und Herausforderungen der Materialflusskostenrechnung Computing Incentives for User-Based Relocation in Carsharing Sustainability’s Coming Home: Preliminary Design Principles for the Sustainable Smart District Substitution of hazardous chemical substances using Deep Learning and t-SNE A Hierarchy of DSMLs in Support of Product Life-Cycle Assessment A Survey of Smart Energy Services for Private Households Door-to-Door Mobility Integrators as Keystone Organizations of Smart Ecosystems: Resources and Value Co-Creation – A Literature Review Ein Entscheidungsunterstützungssystem zur ökonomischen Bewertung von Mieterstrom auf Basis der Clusteranalyse Discovering Blockchain for Sustainable Product-Service Systems to enhance the Circular Economy Digitale Rückverfolgbarkeit von Lebensmitteln: Eine verbraucherinformatische Studie Umweltbewusstsein durch audiovisuelles Content Marketing? Eine experimentelle Untersuchung zur Konsumentenbewertung nachhaltiger Smartphones Towards Predictive Energy Management in Information Systems: A Research Proposal A Web Browser-Based Application for Processing and Analyzing Material Flow Models using the MFCA Methodology Track 13: Digital Work - Social, mobile, smart On Conversational Agents in Information Systems Research: Analyzing the Past to Guide Future Work The Potential of Augmented Reality for Improving Occupational First Aid Prevent a Vicious Circle! The Role of Organizational IT-Capability in Attracting IT-affine Applicants Good, Bad, or Both? Conceptualization and Measurement of Ambivalent User Attitudes Towards AI A Case Study on Cross-Hierarchical Communication in Digital Work Environments ‘Show Me Your People Skills’ - Employing CEO Branding for Corporate Reputation Management in Social Media A Multiorganisational Study of the Drivers and Barriers of Enterprise Collaboration Systems-Enabled Change The More the Merrier? The Effect of Size of Core Team Subgroups on Success of Open Source Projects The Impact of Anthropomorphic and Functional Chatbot Design Features in Enterprise Collaboration Systems on User Acceptance Digital Feedback for Digital Work? Affordances and Constraints of a Feedback App at InsurCorp The Effect of Marker-less Augmented Reality on Task and Learning Performance Antecedents for Cyberloafing – A Literature Review Internal Crowd Work as a Source of Empowerment - An Empirical Analysis of the Perception of Employees in a Crowdtesting Project Track 14: Geschäftsmodelle und digitales Unternehmertum Dividing the ICO Jungle: Extracting and Evaluating Design Archetypes Capturing Value from Data: Exploring Factors Influencing Revenue Model Design for Data-Driven Services Understanding the Role of Data for Innovating Business Models: A System Dynamics Perspective Business Model Innovation and Stakeholder: Exploring Mechanisms and Outcomes of Value Creation and Destruction Business Models for Internet of Things Platforms: Empirical Development of a Taxonomy and Archetypes Revitalizing established Industrial Companies: State of the Art and Success Principles of Digital Corporate Incubators When 1+1 is Greater than 2: Concurrence of Additional Digital and Established Business Models within Companies Special Track 1: Student Track Investigating Personalized Price Discrimination of Textile-, Electronics- and General Stores in German Online Retail From Facets to a Universal Definition – An Analysis of IoT Usage in Retail Is the Technostress Creators Inventory Still an Up-To-Date Measurement Instrument? Results of a Large-Scale Interview Study Application of Media Synchronicity Theory to Creative Tasks in Virtual Teams Using the Example of Design Thinking TrustyTweet: An Indicator-based Browser-Plugin to Assist Users in Dealing with Fake News on Twitter Application of Process Mining Techniques to Support Maintenance-Related Objectives How Voice Can Change Customer Satisfaction: A Comparative Analysis between E-Commerce and Voice Commerce Business Process Compliance and Blockchain: How Does the Ethereum Blockchain Address Challenges of Business Process Compliance? Improving Business Model Configuration through a Question-based Approach The Influence of Situational Factors and Gamification on Intrinsic Motivation and Learning Evaluation von ITSM-Tools für Integration und Management von Cloud-Diensten am Beispiel von ServiceNow How Software Promotes the Integration of Sustainability in Business Process Management Criteria Catalog for Industrial IoT Platforms from the Perspective of the Machine Tool Industry Special Track 3: Demos & Prototyping Privacy-friendly User Location Tracking with Smart Devices: The BeaT Prototype Application-oriented robotics in nursing homes Augmented Reality for Set-up Processe Mixed Reality for supporting Remote-Meetings Gamification zur Motivationssteigerung von Werkern bei der Betriebsdatenerfassung Automatically Extracting and Analyzing Customer Needs from Twitter: A “Needmining” Prototype GaNEsHA: Opportunities for Sustainable Transportation in Smart Cities TUCANA: A platform for using local processing power of edge devices for building data-driven services Demonstrator zur Beschreibung und Visualisierung einer kritischen Infrastruktur Entwicklung einer alltagsnahen persuasiven App zur Bewegungsmotivation für ältere Nutzerinnen und Nutzer A browser-based modeling tool for studying the learning of conceptual modeling based on a multi-modal data collection approach Exergames & Dementia: An interactive System for People with Dementia and their Care-Network Workshops Workshop Ethics and Morality in Business Informatics (Workshop Ethik und Moral in der Wirtschaftsinformatik – EMoWI’19) Model-Based Compliance in Information Systems - Foundations, Case Description and Data Set of the MobIS-Challenge for Students and Doctoral Candidates Report of the Workshop on Concepts and Methods of Identifying Digital Potentials in Information Management Control of Systemic Risks in Global Networks - A Grand Challenge to Information Systems Research Die Mitarbeiter von morgen - Kompetenzen künftiger Mitarbeiter im Bereich Business Analytics Digitaler Konsum: Herausforderungen und Chancen der Verbraucherinformati

A novel therapeutical approach?

Titel und Inhalt i 1. Einleitung 1 2. Zielsetzung 16 3. Material und Methoden 17 4. Ergebnisse 40 5. Diskussion 82 Anhang IBisher sind Etiologie und Pathogenese chronisch entzündlicher Darmerkrankungen (CED), Colitis ulcerosa und Morbus Crohn, nicht vollständig geklärt. Derzeit geht man von einem Zusammenspiel dreier Faktoren aus, die an der Entstehung und der Chronizität der Entzündung beteiligt sind: der genetischen Prädisposition, Umwelteinflüssen und immunologischen Faktoren. Da γδ T-Zellen immun-regulatorisches Potential besitzen und im Tiermodell protektive Effekte bei anderen entzündlichen Erkrankungen wie Arthritis, Diabetes und Lupus zeigten, wurde ihre Rolle bei CED näher untersucht. In verschiedenen CED- Tiermodellen (induzierbare und genetische) konnte in einem ersten Schritt gezeigt werden, dass sich eine Depletion/Defizienz der γδ T-Zellen negativ auf die Entstehung und den Verlauf einer intestinalen Entzündung auswirkt. Infolge dieser γδ T-Zelldepletion/defizienz wiesen die Tiere eine schwerwiegende Schädigung der intestinalen Mukosa auf, zeigten eine erhöhte Sekretion proinflammatorischer und eine erniedrigte Sekretion antiinflammatorischer Zytokine sowie in einigen Fällen eine erhöhte Mortalität. In einem zweiten Schritt wurde der Einfluss eines γδ T-Zelltransfers auf die Entstehung und den Verlauf der CED in einem chemisch-induzierten Tiermodell untersucht. Die transferierten γδ T-Zellen stammten entweder von Wildtyp Mäusen oder wurden aus Interleukin-10 transgenen Tieren isoliert. Unabhängig vom Spender wirkten sich die transferierten γδ T-Zellen protektiv auf die Entstehung und den Verlauf einer chemisch-induzierten Colitis aus. Die Tiere zeigten ein verlängertes Überleben, eine verminderte intestinale Schädigung und eine erhöhte Sekretion antiinflammatorischer Zytokine sowie eine erniedrigte Sekretion proinflammatorischer Zytokine. Der γδ T-Zelltransfer bei etablierter Colitis zeigte zwar keinen Einfluss auf die Mortalität, das mukosale Epithel der Tiere war jedoch weniger geschädigt als das untransferierter Tiere. Gleichzeitig war die Sekretion antiinflammatorischer Zytokine erhöht, die proinflammatorischer Zytokine erniedrigt. Da sich im Tiermodell die Depletion/Defizienz von γδ T-Zellen auf das Entzündungsgeschehen bei CED negativ und der Transfer positiv bzw. protektiv auswirkt, scheinen γδ T-Zellen einen protektiven Effekt auf das mukosale Epithel sowie einen regulatorischen Effekt auf das intestinale Immunsystem auszuüben. Diese Effekte vermitteln sie vermutlich über die Regulation der Zytokindisbalance, die Suppression von unkontrolliert proliferierenden Entzündungszellen und ihre Beteiligung an der epithelialen Regeneration. Dieses Potenzial wurde in vitro näher untersucht. Es zeigte sich, dass humane γδ T-Zellen anerg sind und das Wachstum von CD4+ T-Zellen supprimieren. Diese Suppression üben sie schon in geringen Zellverhältnissen aus. Ferner sezernieren sie vernachlässigbar geringe Mengen an IL-2, jedoch große Mengen an IL-4, IL-10, TGF-β und IFN-γ. So konnte erstmals gezeigt werden, dass γδ T-Zellen sowohl protektives als auch regulatorisches Potenzial besitzen. Diese Eigenschaften machen sie für den therapeutischen Einsatz zur Behandlung von CED interessant. Einerseits proliferieren γδ T-Zellen aufgrund ihrer Anergie nicht unkontrolliert, andererseits können sie die unkontrollierte Ausbreitung autoreaktiver Entzündungszellen hemmen. Durch die Sekretion von Th2-Zytokinen und des Wachstumsfaktors TGF-β sind sie an der epithelialen Regeneration beteiligt und wirken antiinflammatorisch. Die Sekretion von IFN-γ sowie die Regulation der IFN-γ-Produktion durch αβ T-Zellen befähigt sie gleichzeitig zur Infekt- und Tumorabwehr. Daher wurde eine Anreicherung der γδ T-Zellen aus humanen Blutlymphozyten durch verschiedene Stimulationen getestet. Dabei erwiesen sich zwei Stimulationsansätze mit synthetischen Phospho-Antigenen als erfolgversprechend. Zum einen die Stimulation mit Isopentenylpyrophosphat, zum anderen die Stimulation mit Alendronat unter Th2- priming Bedingungen. In beiden Ansätzen war für eine erfolgreiche Anreicherung die Zugabe von rekombinantem IL-2 essentiell. Allerdings müssen diese Anreicherungsansätze noch optimiert werden. Zusätzlich sollte geklärt werden, ob die angereicherten γδ T-Zellen unter der Kultivierung ihren regulatorischen Phänotyp beibehalten haben. Zusammenfassend wird festgestellt, dass γδ T-Zellen einen neuen Ansatzpunkt für die Therapie von CED darstellen, jedoch weiterführende Untersuchungen hinsichtlich ihrer Generierung und ihres therapeutischen Einsatzes erforderlich sind.So far, the etiology and pathogenesis of inflammatory bowel disease (IBD), ulcerative Colitis and Crohn´s Disease, is unclear. At present, three factors, that is genetic predisposition, environmental influences, and immunological factors seem responsible for the development and perpetuation of intestinal inflammation. As γδ T cells possess immunoregulatory potential as well as protective effects in other animal models of inflammatory diseases e.g., arthritis, diabetes und lupus, their role in IBD was examined. First, it was shown in various IBD animal models (inducible and genetic), that the depletion of and deficiency in γδ T cells affected the development and the course of intestinal inflammation negatively. Due to γδ T cell depletion/deficiency the animals displayed a severely damaged intestinal mucosa and increased secretion of proinflammatory as well as decreased secretion of antiinflammatory cytokines with increased mortality in some cases. Second, the influence of γδ T cell transfer on the development and the course of IBD was examined in a chemically induced animal model. The γδ T cells transferred were isolated from wildtype mice or from interleukin-10 transgene animals. These γδ T cells acted independently from their origin protectively on the development and the course of chemically induced colitis. The animals showed increased survival, diminished intestinal injury, and an increased secretion of antiinflammatory cytokines as well as a decreased secretion of proinflammatory cytokines. The γδ T cell transfer at established colitis though not influencing the mortality, resulted in less damaged mucosal epithelium. Simultaneously, the secretion of antiinflammatory cytokines was increased, while the secretion of proinflammatory cytokines was decreased. As the depletion of and the deficiency in γδ cells affected the inflammation in IBD negatively, while the transfer of γδ T cells had a positive and protective effect, respectively, it seems likely, that γδ T cells have a protective effect on the mucosal epithelium as well as a regulatory effect on the intestinal immune system. These effects seem to be mediated by the regulation of the cytokine dysbalance, the suppression of uncontrolled proliferating inflammatory cells, and their involvement in epithelial regeneration. This potential was examined more closely in in vitro experiments. The in vitro data showed, that human γδ T cells are anergic and suppress the growth of CD4+ T cells. They mediate this suppressive effect even at low cell ratios. Additionally, they secrete low amounts of IL-2, but high quantities of IL-4, IL-10, TGF-β, and IFN-γ. The data presented herein showed for the first time, that γδ T cells possess both, protective and regulatory potential. These properties of γδ T cells make them an interesting candidate for IBD immunotherapy. On the one hand, due to their anergy γδ T cells do not proliferate in an autocrine fashion and are able to suppress growth of autoreactive inflammatory cells. Due to their secretion of Th2 cytokines and the growth factor TGF-β, they are involved in epithelial regeneration and act antiinflammatorily. Further, they are able to prevent infections and tumor formation due to their secretion of IFN-γ as well as their regulation of IFN-γ production by αβ T cells. Therefore, various stimulations for the enrichment of γδ T cells from human peripheral blood lymphocytes were tested. Two approaches employing synthetic phosphoantigens seem promising; the stimulation with Isopentenylpyrophosphate and the stimulation with Alendronate under Th2-priming conditions. The addition of endogenous IL-2 was essential for successful enrichment using both methods. Still, the expansion process needs further optimization. Additionally, it is necessary to clarify if the γδ T cells maintain their regulatory phenotype under these culture conditions. In summary, γδ T cells exhibit a new therapeutical approach in treatment of IBD, while further experiments regarding their generation and their therapeutical application are necessary

Murine T-Cell Transfer Colitis as a Model for Inflammatory Bowel Disease.

Inflammatory bowel disease (IBD) is a group of severe chronic inflammatory conditions of the human gastrointestinal tract. Murine models of colitis have been invaluable tools to improve the understanding of IBD development and pathogenesis. While the disease etiology of IBD is complex and multifactorial, CD4+ T helper cells have been shown to strongly contribute to the disease pathogenesis of IBD. Here, we present a detailed protocol of the preclinical model of T-cell transfer colitis, which can easily be utilized in the laboratory to study T helper cell functions in intestinal inflammation

The proteasome inhibitior bortezomib depletes plasma cells and ameliorates clinical manifestations of refractory systemic lupus erythematosus

Objectives To investigate whether bortezomib, a proteasome inhibitor approved for treatment of multiple myeloma, induces clinically relevant plasma cell (PC) depletion in patients with active, refractory systemic lupus erythematosus (SLE). Methods Twelve patients received a median of two (range 1–4) 21-day cycles of intravenous bortezomib (1.3 mg/m2) with the coadministration of dexamethasone (20 mg) for active SLE. Disease activity was assessed using the SLEDAI-2K score. Serum concentrations of anti–double-stranded DNA (anti-dsDNA) and vaccine-induced protective antibodies were monitored. Flow cytometry was performed to analyse peripheral blood B-cells, PCs and Siglec-1 expression on monocytes as surrogate marker for type-I interferon (IFN) activity. Results Upon proteasome inhibition, disease activity significantly declined and remained stable for 6 months on maintenance therapies. Nineteen treatment-emergent adverse events occurred and, although mostly mild to moderate, resulted in treatment discontinuation in seven patients. Serum antibody levels significantly declined, with greater reductions in anti-dsDNA (∼60%) than vaccine-induced protective antibody titres (∼30%). Bortezomib significantly reduced the numbers of peripheral blood and bone marrow PCs (∼50%), but their numbers increased between cycles. Siglec-1 expression on monocytes significantly declined. Conclusions These findings identify proteasome inhibitors as a putative therapeutic option for patients with refractory SLE by targeting PCs and type-I IFN activity, but our results must be confirmed in controlled trials

Pro-inflammatory cytokine responses in ilea of PACAP-treated mice.

<p>Ileitis was induced by peroral infection of mice with <i>T. gondii</i> at day 0. (A) IL-23p19 and (B) IL-22 mRNA expression levels (by RT-PCR) as well as (C) IFN-γ and (D) MCP-1 protein concentrations (by CBA) were determined in <i>ex vivo</i> ileum biopsies obtained from mice following PACAP prophylactic treatment (Proph, white bars), PACAP therapy (Therap, gray bars), or placebo application (black bars) at day 7 post infection as compared to naïve animals (lined bars). mRNA expression levels are expressed as fold changes relative to HPRT mRNA expression (Arbitrary Units). Numbers of analyzed animals are given in parentheses. Mean values, standard errors of the mean, and significance levels as determined by the Mann-Whitney-U test are indicated. Data are pooled from three independent experiments.</p

Intestinal immune cell responses following ileitis induction in PACAP-treated mice.

<p>Ileitis was induced by peroral infection of mice with <i>T. gondii</i> at day 0. The average number of cells positive for (A) CD3 (T lymphocytes), (B) FOXP3 (regulatory T cells, Tregs), (C) MPO7 (myeloperoxidase-7, neutrophils and monocytes), and (D) F4/80 (macrophages) from at least six high power fields (HPF, 400× magnification) per animal were determined microscopically in immunohistochemically stained ileum sections derived from mice following PACAP prophylactic treatment (Proph, white bars), PACAP therapy (Therap, gray bars), or placebo application (black bar) at day 7 post infection as compared to naïve controls (lined bars). Numbers of analyzed animals are given in parentheses. Mean values, standard errors of the mean, and significance levels as determined by the Mann-Whitney-U test are indicated. Data are pooled from three independent experiments.</p

Amelioration of acute ileitis in PACAP-treated mice.

<p>Ileitis was induced by peroral infection of mice with <i>T. gondii</i> at day 0. (A) Relative body weight loss of mice (in %), (B) relative small intestinal shortening (in %), and (C) histopathological scores of the terminal ileum following PACAP prophylactic treatment (Proph, white bars), PACAP therapy (Therap, gray bars) or placebo application (black bar) at day 7 post infection. Numbers of analyzed animals are given in parentheses. Mean values, standard errors of the mean, and significance levels as determined by the Mann-Whitney-U test are indicated. Data are pooled from three independent experiments. (D) Representative photomicrographs (100× magnification; scale bar 100 µm) of histopathological changes in ileal paraffin sections from three independent experiments are shown. Naïve, uninfected mice served as controls.</p