Phenotyping of Fecal Microbiota of Winnie, a Rodent Model of Spontaneous Chronic Colitis, Reveals Specific Metabolic, Genotoxic, and Pro-inflammatory Properties

Abstract Winnie, a mouse carrying a missense mutation in the MUC2 mucin gene, is a valuable model for inflammatory bowel disease (IBD) with signs and symptoms that have multiple similarities with those observed in patients with ulcerative colitis. MUC2 mucin is present in Winnie, but is not firmly compacted in a tight inner layer. Indeed, these mice develop chronic intestinal inflammation due to the primary epithelial defect with signs of mucosal damage, including thickening of muscle and mucosal layers, goblet cell loss, increased intestinal permeability, enhanced susceptibility to luminal inflammation-inducing toxins, and alteration of innervation in the distal colon. In this study, we show that the intestinal environment of the Winnie mouse, genetically determined by MUC2 mutation, selects an intestinal microbial community characterized by specific pro-inflammatory, genotoxic, and metabolic features that could imply a direct involvement in the pathogenesis of chronic intestinal inflammation. We report results obtained by using a variety of in vitro approaches for fecal microbiota functional characterization. These approaches include Caco-2 cell cultures and Caco-2/THP-1 cell co-culture models for evaluation of geno-cytotoxic and pro-inflammatory properties using a panel of 43 marker RNAs assayed by RT-qPCR, and cell-based phenotypic testing for metabolic profiling of the intestinal microbial communities by Biolog EcoPlates. While adding a further step towards understanding the etiopathogenetic mechanisms underlying IBD, the results of this study provide a reliable method for phenotyping gut microbial communities, which can complement their structural characterization by providing novel functional information

Epidermal Growth Factor Receptor Intron-1 Polymorphism Predicts Gefitinib Outcome in Advanced Non-small Cell Lung Cancer

IntroductionEpidermal growth factor receptor (EGFR) gene intron 1 contains a polymorphic single sequence dinucleotide repeat (CA)n whose length has been found to inversely correlate with transcriptional activity. This study was designed to assess the role of (CA)n polymorphism in predicting the outcome of gefitinib treatment in advanced non-small cell lung cancer (NSCLC).MethodsBlood and tumor tissue from 58 patients with advanced NSCLC submitted to gefitinib were collected. EGFR intron 1 gene polymorphism, along with EGFR gene mutation, gene copy number and immunohistochemistry expression were determined. Moreover, a panel of lung cancer cell lines characterized for EGFR intron 1 polymorphism was also studied.ResultsEGFR intron 1 polymorphism showed a statistically significant correlation with the gefitinib response (response rate 25 versus 0%, for patients with a (CA)16 and with a (CA)else genotype, respectively; p = 0.044). Patients with a (CA)16 genotype had a longer survival compared with those with a (CA)else genotype (11.4 versus 4.8 months, respectively; p = 0.037). In addition, cell lines lacking the (CA)16 allele showed a statistically significant higher IC50 compared with cell lines bearing at least one (CA)16 allele (p = 0.003).ConclusionsThis study supports a potential role of EGFR intron 1 polymorphism in predicting the outcome of gefitinib treatment in advanced NSCLC

Characterization of age-dependent immune signatures after Influenza vaccination

Die fortschreitende Alterung der Bevölkerung stellt die Medizin vor zahlreiche komplexe Herausforderungen. Neben Verbesserungen im therapeutischen Bereich stehen prophylaktische Maßnahmen mit positivem Kosten-Nutzen-Verhältnis im Fokus eines leistungsstarken Gesundheitssystems. Impfungen sind ein Beispiel prophylaktischer Ansätze, die Morbidität und Mortalität erheblich beeinflussen können. Weil Impfungen im Alter aber an Effektivität verlieren, kommt dem Verständnis für die Mechanismen dieser sogenannten Immunoseneszenz eine große Bedeutung zu. Ziel dieser Arbeit war die Identifikation immunologischer Signaturen, die einen Einfluss auf die altersabhängig veränderte Immunität ausüben. Die saisonale Influenza-Impfung weist eine besonders hohe Rate an Impfversagen bei älteren Menschen auf und stellt daher ein gutes Modell für die Untersuchung altersabhängiger Unterschiede in der Immunreaktion nach Vakzinierung dar. In der Studie Protective Immunity in Ageing (PRIMAGE) wurden zwei Alterskohorten (18-30 Jahre vs. 55-65 Jahre) mit dem trivalenten inaktivierten Vakzin (TIV) Mutagrip® 2011/2012 geimpft. Die Impfantwort wurde durch serologische Test mittels Hämagglutinin-Inhibition (HI) – Assay sowie durch die Charakterisierung von B-Lymphozyten mittels Durchflusszytometrie analysiert. Zudem wurden T-Lymphozyten anhand ihres Differenzierungsgrades phänotypisiert und Antigen-spezifische T-Lymphozyten für das Cytomegalie-Virus (CMV) nach in vitro Stimulation charakterisiert. Unseren Daten belegen, dass die Immunantwort bei älteren Menschen geringer ausfällt als bei den Jüngeren, was sich durch verminderte HI-Titer für Influenza H1N1 als auch durch eine geringere Plasmablasten-Expansion darstellte. Zudem fand sich eine im Alter zunehmende relative Anreicherung von Gedächtniszellen im Vergleich zu naiven T-Lymphozyten sowie eine Zunahme an CD4+ und CD8+ T-Lymphozyten, die Erschöpfungsmarkern exprimieren. Wir konnten einen Zusammenhang zwischen dieser Verschiebung des T-Zell-Phänotyps mit dem Vorliegen latenter CMV- Infektionen feststellen. Außerdem wiesen die Antigen-spezifischen T-Lymphozyten in Abhängigkeit ihrer Spezifität für die CMV-Peptide IE-1 und pp65 Unterschiede im Zytokinprofil sowie im Phänotyp auf. Diese Arbeit erlaubt somit Einblicke in die Mechanismen, die dem Phänomen der Immunoseneszenz zugrunde liegen. Neben einer verminderten humoralen Impfantwort wurde ein Zusammenhang dieser Mechanismen mit dem CMV-Status nachgewiesen. Diese Erkenntnisse liefern Argumente dafür, bei der Indikationsstellung zur Immunisierung neben dem anagraphischen auch das immunologische Alter sowie den CMV-Status zu berücksichtigen.Demographics of an ageing society will pose significant challenges to present day health care systems. Along with advances in therapy, improvement of prophylactic medicine with favourable cost benefit analysis is urgently needed. Vaccinations are an example for prophylactic approaches capable of significantly reducing morbidity and mortality. As vaccinations are known to be less efficient in age, many efforts have been made to detect underlying mechanisms of what is called immunosenescence. This study aims at identifying immunological signatures that modulate age-dependent immunity. Flu vaccination has been described to be less efficient in the elderly and is therefore a good model to study the effects of immunosenescence. In our study Protective Immunity in Ageing (PRIMAGE) we vaccinated two age groups with the trivalent inactivated vaccine (TIV) Mutagrip® 2011/2012 and characterised their immune responses. Both serological and cellular responses were determined by either hemagglutinin inhibition assay (HIA) or flow cytometry. Furthermore, we analysed T cell subsets according to their differentiation and specificity towards cytomegalovirus (CMV) after in vitro stimulation. Our data confirms a weaker immune response in the elderly in terms of impaired HI-titer (H1N1) as well as decreased plasmablast-expansion. Age-dependent differences in the T cell compartment were demonstrated by a phenotype shift from naïve to memory and an increase of T cells expressing exhaustion markers in the elderly. In an effort to describe environmental factors in the genesis of these changes, we analysed the CMV status of our donors and found it to be associated with a memory like T cell phenotype as well as exhaustion. We also assessed differential effects in cytokine profile and phänotype depending on T cell specificity towards the two CMV peptides IE-1 and pp65. In summary, this study sheds further light onto the underlying mechanisms of immunosenescence. We could demonstrate an impaired immune response after flu vaccination in the elderly. Ageing as well as CMV infections modulate T cell subsets and may have important implications for the outcome of vaccination. Our data supports the integration of biological age, immunological age and CMV status to further increase the efficacy of vaccination

Highly Predictive Model for a Protective Immune Response to the A(H1N1)pdm2009 Influenza Strain after Seasonal Vaccination

Understanding the immune response after vaccination against new influenza strains is highly important in case of an imminent influenza pandemic and for optimization of seasonal vaccination strategies in high risk population groups, especially the elderly. Models predicting the best sero-conversion response among the three strains in the seasonal vaccine were recently suggested. However, these models use a large number of variables and/or information post- vaccination. Here in an exploratory pilot study, we analyzed the baseline immune status in young

FGFR as potential target in the treatment of squamous non small cell lung cancer

To date therapeutic options for squamous cell lung cancer patients remain scarce because no druggable targets have been identified so far. Aberrant signaling by FGFs (fibroblast growth factors) and FGFRs (fibroblast growth factors receptors) has been implicated in several human cancers and, particularly, in squamous non-small cell lung cancer (NSCLC). FGFR gene amplifications, somatic missense mutations, chromosomal translocations are the most frequent mechanisms able to induce aberrant activation of this pathway. Data from literature have established that the presence of an aberrant FGFR signaling has to be considered a possible negative prognostic factor but predictive of potential sensitivity to FGFR inhibitors. In the last years, clinical research efforts allowed to identify and evaluate promising FGFR inhibitors, such as monoclonal antibodies, ligand traps, non-selective or selective tyrosine kinase inhibitors. This review summarizes the current knowledge about FGFR alterations in NSCLC and the relative inhibitors in development, in particular in squamous NSCLC

Fingerprint Change: Not Visible, But Tangible

Handâfoot syndrome, a chemotherapy-induced cutaneous toxicity, can cause an alteration in fingerprints causing a setback for cancer patients due to the occurrence of false rejections. A colon cancer patient was fingerprinted after not having been able to use fingerprint recognition devices after 6 months of adjuvant chemotherapy. The fingerprint images were digitally processed to improve fingerprint definition without altering the papillary design. No evidence of skin toxicity was present. Two months later, the situation returned to normal. The fingerprint evaluation conducted on 15 identification points highlighted the quantitative and qualitative fingerprint alteration details detected after the end of chemotherapy and 2 months later. Fingerprint alteration during chemotherapy has been reported, but to our knowledge, this particular case is the first ever reported without evident clinical signs. Alternative fingerprint identification methods as well as improved biometric identification systems are needed in case of unexpected situations