Kinetics and cellular site of glycolipid loading control

CD1d-restricted natural killer T cells (NKT cells) possess a wide range of effector and regulatory activities that are related to their ability to secrete both T helper 1 (Th1) cell- and Th2 cell-type cytokines. We analyzed presentation of NKT cell activating α galactosylceramide (αGalCer) analogs that give predominantly Th2 cell-type cytokine responses to determine how ligand structure controls the outcome of NKT cell activation. Using a monoclonal antibody specific for αGalCer-CD1d complexes to visualize and quantitate glycolipid presentation, we found that Th2 cell-type cytokinebiasing ligands were characterized by rapid and direct loading of cell-surface CD1d proteins. Complexes formed by association of these Th2 cell-type cytokine-biasing αGalCer analogs with CD1d showed a distinctive exclusion from ganglioside-enriched, detergent-resistant plasma membrane microdomains of antigen-presenting cells. These findings help to explain how subtle alterations in glycolipid ligand structure can control the balance of proinflammatory and antiinflammatory activities of NKT cells

The role of Fusobacterium nucleatum and the Epstein-Barr virus encoded EBV-miR-BART10-3p in colorectal cancer - focusing on the genes CXCL8, CSF2, SAP18 and CCND1

Kolorektal kreft er den tredje mest vanlige krefttypen i verden, og den nest ledende årsaken til dødsfall forårsaket av kreft for begge kjønn. Utviklingen av kolorektal kreft er en prosess som foregår gjennom flere trinn som involverer både genetiske og epigenetiske endringer i tumorsuppressorgener og onkogener, som er nødvendige for initiering og progresjon av kreft. Minoriteten av kolorektal kreft-pasienter har familiær eller arvelig form av kolorektal kreft, mens majoriteten av krefttilfellene oppstår sporadisk, noe som antyder at miljø og livsstil representerer risikofaktorer for kolorektal kreft. Økt bevissthet omkring tarmmikrobiota sin rolle i initieringen, tilrettelegging og utvikling av kolorektal kreft har i senere år vokst frem. Epstein-Barr virus og den orale bakterien Fusobacterium nucelatum er påvist oppregulert i kolorektalt tumorvev sammenlignet med tilstøtende friskt vev, noe som fremhever deres rolle i initiering og progresjon av kolorektal kreft. Fusobacterium nucleatum er blitt vist å infisere og internalisere i den kolorektale cellelinjen DLD-1, visualisert ved LSM-konfokal mikroskopi og fluorescens-farging, samt å oppregulere cytokinene CXCL8 og CSF2 i co-kultur i cellelinjene DLD-1 og SW620. Genekspresjonen til CXCL8 og CSF2 og protein sekresjonen av CXCL8, ble vist å øke både ved inkubasjonstid og økt konsentrasjon av F. nucleatum, ved hjelp av real-time qPCR og ELISA. CXCL8 og CSF2 er begge involvert i kroppen sin immunrespons, og er vist å være involvert i initieringen og progresjonen av kolorektal kreft ved å skape et inflammatorisk mikromiljø som er gunstig for tumorprogresjonen. Vi viser at EBV-miR-BART10-3p binder og nedregulerer genene SAP18 og CCND1 i cellelinjen SW620 ved real-time qPCR og luciferase analyse. En pasient-derivert sfæroide metode (CTOSs), basert på CTOSs-metoden fra Kondo et al. (2011) og Jeppesen et al. (2017) er blitt utviklet og etablert, i form av sfæroidedyrking, splitting, frysing og tining. CTOSs-metoden gir et høyt utbytte av levedyktige sfæroider, som overlever prosessen med splitting, frysing og tining

Child Safety Signal Learning in the Context of Parental Worry, Overprotection, PTSD, and Depression

Thesis (Ph.D.)--University of Washington, 2016-08Individuals with anxiety-related psychopathology often have impairments in safety signal learning, which is thought to be a biomarker of anxiety-related disorders, and which manifests in difficulty inhibiting fear to safety cues (e.g., Jovanovic et al. 2010). Accordingly, one way in which parental anxiety-related psychopathology may affect a child’s development is by changing the nature of pivotal, early parent-child learning experiences related to identifying what is safe and what is not. This study used the well-established conditional discrimination (AX+/BX-) paradigm to explore safety signal learning in 8-11 year-old children in relation to parental worry, overprotection, PTSD, and depression. This paradigm assesses fear-potentiated startle (FPS) to conditioned stimuli (CS) that are either paired with an aversive air puff to the throat (AX+), or that are never paired with the air puff (BX-). FPS to the threat cue (A) presented in conjunction with the safety cue (B) in the absence of the air puff (AB) serves as the critical test of safety signal learning. Dependent variables were FPS to the AX+ and AB trials, as well as retrospective expectancy ratings of whether the air puff had been paired with each CS. Children whose parents were higher in overprotective parenting had higher FPS to the safety transfer cue (AB) than to the safety cue, suggesting impaired safety signal learning. Additionally, higher parental depression was associated with lower initial child FPS to both the danger and safety cues, suggesting a blunting effect of parental depression on child fear responding. By contrast, children whose parents reported higher symptoms of worry did not differ from children whose parents reported lower symptoms of worry in FPS to the danger, safety, or safety transfer cues. Parental PTSD was also not strongly associated with child fear responding or inhibition. Further, children whose parents were lower and higher in worry or overprotective parenting did not differ in cognitively discriminating among the danger, safety, and safety transfer cues. This study provides valuable information about fear inhibition during the middle childhood years, suggesting that impairments in safety signal learning are specifically related to overprotective parenting. Overprotective parenting may thus be a viable target for the prevention and treatment of anxiety-related disorders in children

Emotion Regulation in Adults with a History of Childhood Abuse Following PTSD Treatment

Thesis (Master's)--University of Washington, 2012Objective: The findings that adults with a history of childhood abuse (CA) have greater emotion regulation difficulties than adult trauma survivors without a history of CA (e.g., Cloitre, Scarvalone, & Difede, 1997) has led to the hypothesis that existing PTSD treatments may not be adequate for improving the emotion regulation deficits of adults with a history of CA (Cloitre et al., 2010). This study compared changes in emotion regulation over the course of PTSD treatment with either prolonged exposure (PE) therapy or sertraline in adults with and without a history of CA. Method: Two hundred adults with PTSD received 10 weeks of either PE or sertraline. Emotion regulation and trait affect were assessed pre- and post-treatment with the Emotion Regulation Questionnaire (Gross & John, 2003), the Negative Mood Regulation Scale (Catanzaro & Mearns, 1990), and the Positive and Negative Affect Scale (Watson, Clark, & Tellegen, 1988). Results: Individuals with a history of CA did not differ from individuals without a history of CA at pre-treatment on PTSD severity, emotion regulation, or positive/negative affect. In addition, treatment was effective at improving emotion regulation and trait affect in those with and without a history of CA, and no significant differences in emotion regulation or trait affect emerged at post-treatment between adults with and without a history of CA. Furthermore, non-inferiority analyses indicated that the emotion regulation and trait affect outcomes of those with a history of CA were not inferior to the emotion regulation and trait affect outcomes of those without a history of CA. Conclusion: Contrary to clinical lore regarding clients presenting for PTSD treatment, the current findings suggest that those with a history of CA do not differ from those without a history of CA in terms of emotion regulation and trait affect. Further, these findings cast doubt on the assumption that CA predicts worse emotion regulation outcomes following PTSD treatment

Pharmacological treatment of anxiety disorders: Current treatments and future directions

Modern pharmacological treatments for anxiety disorders are safer and more tolerable than they were 30 years ago. Unfortunately, treatment efficacy and duration have not improved in most cases despite a greater understanding of the pathophysiology of anxiety. Moreover, innovative treatments have not reached the market despite billions of research dollars invested in drug development. In reviewing the literature on current treatments, we argue that evidence-based practice would benefit from better research on the causes of incomplete treatment response as well as the comparative efficacy of drug combinations and sequencing. We also survey three approaches to the development of innovative anxiety treatments: the continued development of drugs based on specific neuroreceptors; the pharmacologigcal manipulation of fear-related memory; and the electrical or electromagnetic stimulation of specific brain areas. We highlight directions for future research, as none of these approaches is ready for clinical use. This record was migrated from the OpenDepot repository service in June, 2017 before shutting down

Improved outcomes in NOD mice treated with a novel Th2 cytokine-biasing NKT cell activator

Abstract Activation of CD1d-restricted invariant NKT (iNKT) cells by α-galactosylceramide (αGalCer) significantly suppresses development of diabetes in NOD mice. The mechanisms of this protective effect are complex, involving both Th1 and Th2 cytokines and a network of regulatory cells including tolerogenic dendritic cells. In the current study, we evaluated a newly described synthetic αGalCer analog (C20:2) that elicits a Th2-biased cytokine response for its impact on disease progression and immunopathology in NOD mice. Treatment of NOD mice with αGalCer C20:2 significantly delayed and reduced the incidence of diabetes. This was associated with significant suppression of the late progression of insulitis, reduced infiltration of islets by autoreactive CD8+ T cells, and prevention of progressive disease-related changes in relative proportions of different subsets of dendritic cells in the draining pancreatic lymph nodes. Multiple favorable effects observed with αGalCer C20:2 were significantly more pronounced than those seen in direct comparisons with a closely related analog of αGalCer that stimulated a more mixed pattern of Th1 and Th2 cytokine secretion. Unlike a previously reported Th2-skewing murine iNKT cell agonist, the αGalCer C20:2 analog was strongly stimulatory for human iNKT cells and thus warrants further examination as a potential immunomodulatory agent for human disease.</jats:p