The role of the inflammasome in the respiratory innate immune response to viruses and pollutants: insights for asthma pathogenesis

The respiratory mucosal innate immune system is composed of both structural cells, such as airway epithelial cells, and immune cells, such as macrophages, that together determine the appropriate immune response to inhaled stimuli. These cells use pattern recognition receptors to distinguish between harmless and harmful stimuli by recognizing conserved microbial pathogen-associated molecular patterns (PAMPs) and endogenously derived damage-associated molecular patterns (DAMPs) from damaged tissue. Inappropriate immune responses to normally innocuous stimuli underpin the pathogenesis of a number of immune disorders, including asthma, a chronic inflammatory disease of the airway typified by difficulty breathing in response to a trigger. In this dissertation, we explored the contribution of the inflammasome signaling complex to respiratory mucosal host defense against two sources of asthma exacerbation: influenza A virus infection and inhalation of the oxidant air pollutant ozone (O3). The inflammasome is an innate immune complex composed of a pattern recognition receptor, the protease caspase-1, and an adaptor protein (PYCARD) that when formed induces activation of caspase-1-mediated processing of the pro-inflammatory mediators IL-1B and IL-18 or cell death. Our results show that caspase-1 and the inflammasome contribute to the airway epithelial cell innate immune response to influenza and that this pathway is modified by asthma, suggesting a role for caspase-1 in virus-induced asthma exacerbation. Using a mouse model of allergic airway inflammation, we also show that caspase-1 may be involved in the development of allergic asthma, though its function in asthma development is complex. In the context of O3, we found increased presence of several DAMPs that may activate inflammasome signaling in the airway of healthy volunteers exposed to O3, but no evidence to suggest that inflammasome signaling strongly contributes to the innate immune response to O3. Finally, we identified a mechanism by which O3 alters the interaction between epithelial cells and macrophages leading to the accumulation of DAMPs that may activate innate immune responses in the lung. In summary, our results shed light on the function of the inflammasome in the human respiratory innate immune response to viruses and pollutants, and provide insight on the contribution of inflammasome signaling to asthma pathogenesis.Doctor of Philosoph

Assessing the risk of venous thromboembolic events in women taking progestin-only contraception: a meta-analysis

Objectives: To evaluate the risk of venous thromboembolic events associated with the use of progestin-only contraception and whether that risk differs with the mode of drug delivery (oral, intrauterine, or depot injection). Design: Systematic review and meta-analysis of randomised controlled trials and observational studies. Data sources: Pubmed, Embase, Cochrane Library, and reference lists of relevant reviews. Study selection: Randomised controlled trials and case-control, cohort, and cross sectional studies with venous thromboembolic outcome for progestin-only contraception reported relative to a non-hormone comparator group. Data extraction: Data were extracted by two independent investigators, and consensus for inclusion was reached after assessment by additional investigators. Results: Among the 2022 unique references identified by all searches, eight observational studies fulfilled inclusion criteria. A total of 147 women across all studies were diagnosed with a venous thromboembolic event while taking progestin-only contraception, and the summary measure for the adjusted relative risk of a venous thromboembolic episode for users versus non-users of a progestin-only contraceptive was, based on the random effects model, 1.03 (95% CI 0.76 to 1.39). Subgroup analysis confirmed there was no association between venous thromboembolic risk and progestin-only pills (relative risk 0.90 (0.57 to 1.45)) or a progestin intrauterine device (0.61 (0.24 to 1.53)). The relative risk of a venous thromboembolic event for users of an injectable progestin versus non-users was 2.67 (1.29 to 5.53). Conclusions: Published data assessing the risk of venous thromboembolism in women prescribed progestin-only contraception are limited. In this meta-analysis of eight observational studies, the use of progestin-only contraception was not associated with an increased risk of venous thromboembolism compared with non-users of hormonal contraception. The potential association between injectable progestins and thrombosis requires further study

Effects of air pollutants on innate immunity: The role of Toll-like receptors and nucleotide-binding oligomerization domain–like receptors

Interactions between exposure to ambient air pollutants and respiratory pathogens have been shown to modify respiratory immune responses. Emerging data suggest key roles for Toll-like receptor (TLR) and nucleotide-binding oligomerization domain–like receptor (NLR) signaling in pathogen-induced immune responses. Similarly, immune responses elicited by exposure to air pollutants are mediated by specific TLR- and NLR-dependent mechanisms. This review article will summarize current knowledge about how air pollutants modify TLR- and NLR-dependent signaling and host defense responses in the lung

Interaction with Epithelial Cells Modifies Airway Macrophage Response to Ozone

The initial innate immune response to ozone (O3) in the lung is orchestrated by structural cells, such as epithelial cells, and resident immune cells, such as airway macrophages (Macs). We developed an epithelial cell–Mac coculture model to investigate how epithelial cell–derived signals affect Mac response to O3. Macs from the bronchoalveolar lavage (BAL) of healthy volunteers were cocultured with the human bronchial epithelial (16HBE) or alveolar (A549) epithelial cell lines. Cocultures, Mac monocultures, and epithelial cell monocultures were exposed to O3 or air, and Mac immunophenotype, phagocytosis, and cytotoxicity were assessed. Quantities of hyaluronic acid (HA) and IL-8 were compared across cultures and in BAL fluid from healthy volunteers exposed to O3 or air for in vivo confirmation. We show that Macs in coculture had increased markers of alternative activation, enhanced cytotoxicity, and reduced phagocytosis compared with Macs in monoculture that differed based on coculture with A549 or 16HBE. Production of HA by epithelial cell monocultures was not affected by O3, but quantities of HA in the in vitro coculture and BAL fluid from volunteers exposed in vivo were increased with O3 exposure, indicating that O3 exposure impairs Mac regulation of HA. Together, we show epithelial cell–Mac coculture models that have many similarities to the in vivo responses to O3, and demonstrate that epithelial cell–derived signals are important determinants of Mac immunophenotype and response to O3

Ozone-derived Oxysterols Affect Liver X Receptor (LXR) Signaling: A POTENTIAL ROLE FOR LIPID-PROTEIN ADDUCTS

When inhaled, ozone (O3) interacts with cholesterols of airway epithelial cell membranes or the lung-lining fluid, generating chemically reactive oxysterols. The mechanism by which O3-derived oxysterols affect molecular function is unknown. Our data show that in vitro exposure of human bronchial epithelial cells to O3 results in the formation of oxysterols, epoxycholesterol-α and -β and secosterol A and B (Seco A and Seco B), in cell lysates and apical washes. Similarly, bronchoalveolar lavage fluid obtained from human volunteers exposed to O3 contained elevated levels of these oxysterol species. As expected, O3-derived oxysterols have a pro-inflammatory effect and increase NF-κB activity. Interestingly, expression of the cholesterol efflux pump ATP-binding cassette transporter 1 (ABCA1), which is regulated by activation of the liver X receptor (LXR), was suppressed in epithelial cells exposed to O3. Additionally, exposure of LXR knock-out mice to O3 enhanced pro-inflammatory cytokine production in the lung, suggesting LXR inhibits O3-induced inflammation. Using alkynyl surrogates of O3-derived oxysterols, our data demonstrate adduction of LXR with Seco A. Similarly, supplementation of epithelial cells with alkynyl-tagged cholesterol followed by O3 exposure causes observable lipid-LXR adduct formation. Experiments using Seco A and the LXR agonist T0901317 (T09) showed reduced expression of ABCA1 as compared with stimulation with T0901317 alone, indicating that Seco A-LXR protein adduct formation inhibits LXR activation by traditional agonists. Overall, these data demonstrate that O3-derived oxysterols have pro-inflammatory functions and form lipid-protein adducts with LXR, thus leading to suppressed cholesterol regulatory gene expression and providing a biochemical mechanism mediating O3-derived formation of oxidized lipids in the airways and subsequent adverse health effects

Individual and group-level job resources and their relationships with individual work engagement

OBJECTIVES: This study adds a multilevel perspective to the well-researched individual-level relationship between job resources and work engagement. In addition, we explored whether individual job resources cluster within work groups because of a shared psychosocial environment and investigated whether a resource-rich psychosocial work group environment is beneficial for employee engagement over and above the beneficial effect of individual job resources and independent of their variability within groups. METHODS: Data of 1,219 employees nested in 103 work groups were obtained from a baseline employee survey of a large stress management intervention project implemented in six medium and large-sized organizations in diverse sectors. A variety of important job resources were assessed and grouped to an overall job resource factor with three subfactors (manager behavior, peer behavior, and task-related resources). Data were analyzed using multilevel random coefficient modeling. RESULTS: The results indicated that job resources cluster within work groups and can be aggregated to a group-level job resources construct. However, a resource-rich environment, indicated by high group-level job resources, did not additionally benefit employee work engagement but on the contrary, was negatively related to it. CONCLUSIONS: On the basis of this unexpected result, replication studies are encouraged and suggestions for future studies on possible underlying within-group processes are discussed. The study supports the presumed value of integrating work group as a relevant psychosocial environment into the motivational process and indicates a need to further investigate emergent processes involved in aggregation procedures across levels

Correction: Effect of Broccoli Sprouts on Nasal Response to Live Attenuated Influenza Virus in Smokers: A Randomized, Double-Blind Study

BackgroundSmokers have increased susceptibility and altered innate host defense responses to influenza virus infection. Broccoli sprouts are a source of the Nrf2 activating agentsulforaphane, and short term ingestion of broccoli sprout homogenates (BSH) has been shown to reduce nasal inflammatory responses to oxidant pollutants.ObjectivesAssess the effects of BSH on nasal cytokines, virus replication, and Nrf2-dependent enzyme expression in smokers and nonsmokers.MethodsWe conducted a randomized, double-blind, placebo-controlled trial comparing the effects of BSH on serially sampled nasal lavage fluid (NLF) cytokines, viral sequence quantity, and Nrf2-dependent enzyme expression in NLF cells and biopsied epithelium. Healthy young adult smokers and nonsmokers ingested BSH or placebo (alfalfa sprout homogenate) for 4 days, designated Days -1, 0, 1, 2. On Day 0 they received standard vaccine dose of live attenuated influenza virus (LAIV) intranasally. Nasal lavage fluids and nasal biopsies were collected serially to assess response to LAIV.ResultsIn area under curve analyses, post-LAIV IL-6 responses (P = 0.03) and influenza sequences (P = 0.01) were significantly reduced in NLF from BSH-treated smokers, whileNAD(P)H: quinoneoxidoreductasein NLF cells was significantly increased. In nonsmokers, a similar trend for reduction in virus quantity with BSH did not reach statistical significance.ConclusionsIn smokers, short term ingestion of broccoli sprout homogenates appears to significantly reduce some virus-induced markers of inflammation, as well as reducing virus quantity. Nutritional antioxidant interventions have promise as a safe, low-cost strategy for reducing influenza risk among smokers and other at risk populations.Trial RegistrationClinicalTrials.gov NCT0126972

The genomes of two key bumblebee species with primitive eusocial organization

Background: The shift from solitary to social behavior is one of the major evolutionary transitions. Primitively eusocial bumblebees are uniquely placed to illuminate the evolution of highly eusocial insect societies. Bumblebees are also invaluable natural and agricultural pollinators, and there is widespread concern over recent population declines in some species. High-quality genomic data will inform key aspects of bumblebee biology, including susceptibility to implicated population viability threats. Results: We report the high quality draft genome sequences of Bombus terrestris and Bombus impatiens, two ecologically dominant bumblebees and widely utilized study species. Comparing these new genomes to those of the highly eusocial honeybee Apis mellifera and other Hymenoptera, we identify deeply conserved similarities, as well as novelties key to the biology of these organisms. Some honeybee genome features thought to underpin advanced eusociality are also present in bumblebees, indicating an earlier evolution in the bee lineage. Xenobiotic detoxification and immune genes are similarly depauperate in bumblebees and honeybees, and multiple categories of genes linked to social organization, including development and behavior, show high conservation. Key differences identified include a bias in bumblebee chemoreception towards gustation from olfaction, and striking differences in microRNAs, potentially responsible for gene regulation underlying social and other traits. Conclusions: These two bumblebee genomes provide a foundation for post-genomic research on these key pollinators and insect societies. Overall, gene repertoires suggest that the route to advanced eusociality in bees was mediated by many small changes in many genes and processes, and not by notable expansion or depauperation