Live Attenuated Chimeric Yellow Fever Dengue Type 2 (Chimeri Vax -DEN2) Vaccine: Phase 1 clinical trial for safety and immunogenicity

A randomized double-blind Phase I Trial was conducted to evaluate safety, tolerability, and immunogenicity of a yellow fever (YF)-dengue 2 (DEN2) chimera (ChimeriVax™-DEN2) in comparison to that of YF vaccine (YF-VAX®). Forty-two healthy YF naïve adults randomly received a single dose of either ChimeriVax™-DEN2 (high dose, 5 log plaque forming units [PFU] or low dose, 3 log PFU) or YF-VAXâ by the subcutaneous route (SC). To determine the effect of YF pre-immunity on the ChimeriVaxTM-DEN2 vaccine, 14 subjects previously vaccinated against YF received a high dose of ChimeriVax™-DEN2 as an open-label vaccine. Most adverse events were similar to YF-VAX® and of mild to moderate intensity, with no serious side-effects. One hundred percent and 92.3% of YF naïve subjects inoculated with 5.0 and 3.0 log10 PFU of ChimeriVaxTM-DEN2, respectively, seroconverted to wt DEN2 (strain 16681); 92% of subjects inoculated with YF-VAX® seroconverted to YF 17D virus but none of YF naïve subjects inoculated with ChimeriVax-DEN2 seroconverted to YF 17D virus. Low seroconversion rates to heterologous DEN serotypes 1, 3, and 4 were observed in YF naïve subjects inoculated with either ChimeriVax™-DEN2 or YF-VAX®. In contrast, 100% of YF immune subjects inoculated with ChimeriVax™-DEN2 seroconverted to all 4 DEN serotypes. Surprisingly, levels of neutralizing antibodies to DEN 1, 2, and 3 viruses in YF immune subjects persisted after 1 year. These data demonstrated that 1) the safety and immunogenicity profile of the ChimeriVax™-DEN2 vaccine is consistent with that of YF-VAX®, and 2) pre-immunity to YF virus does not interfere with ChimeriVaxTM-DEN2 immunization, but induces a long lasting and cross neutralizing antibody response to all 4 DEN serotypes. The latter observation can have practical implications toward development of a dengue vaccine.

The onset and effectiveness of Adult Plant Resistance (APR) in Tallon barley

Field observations of net blotch epidemics indicated that Tallon barley was quite resistant to infection during later stages of growth despite being susceptible as a seedling. A glasshouse experiment was conducted to determine the effectiveness of this resistance and when it became operative. Three cultivars – Gilbert (very susceptible), Patty (resistant) and Tallon – were inoculated at various stages of growth with conidia of Pyrenophora teres f. teres and the infection response and leaf area diseased, recorded 13 days later. The response of Tallon clearly changed from susceptible to moderately susceptible at growth stage 33. Plants sown two weeks earlier were susceptible and plants sown two weeks later were moderately resistant. The response of the other two cultivars at similar growth stages paralleled their seedling responses. The resistance of Tallon appeared to increase with maturity so that, at its most resistant growth stage, the leaf area diseased was just 10% that of the susceptible, Gilbert. While this resistance appears pathotype specific, this experiment demonstrated very effective APR to net blotch. As most losses to this disease occur during the later stages of plant development, APR offers a valuable source of resistance.

Modelling Apoptotic cell clearance within the atherosclerotic plaque

The aged population have an increased susceptibility to infection, therefore function of the innate immune system may be impaired as we age. Macrophages, and their precursors monocytes, play an important role in host defence in the form of phagocytosis, and also link the innate and adaptive immune system via antigen presentation. Classically-activated 'M1' macrophages are pro-inflammatory, which can be induced by encountering pathogenic material or pro-inflammatory mediators. Alternatively activated 'M2' macrophages have a largely reparative role, including clearance of apoptotic bodies and debris from tissues. Despite some innate immune receptors being implicated in the clearance of apoptotic cells, the process has been observed to have a dominant anti-inflammatory phenotype with cytokines such as IL-10 and TGF-ß being implicated. The atherosclerotic plaque contains recruited monocytes and macrophages, and is a highly inflammatory environment despite high levels of apoptosis. At these sites, monocytes differentiate into macrophages and gorge on lipoproteins, resulting in formation of 'foam cells' which then undergo apoptosis, recruiting further monocytes. This project seeks to understand why, given high levels of apoptosis, the plaque is a pro-inflammatory environment. This phenomenon may be the result of the aged environment or an inability of foam cells to elicit an anti-inflammatory effect in response to dying cells. Here we demonstrate that lipoprotein treatment of macrophages in culture results in reduced capacity to clear apoptotic cells. The effect of lipoprotein treatment on apoptotic cell-mediated immune modulation of macrophage function is currently under study.

Influence of iron on bacterial infections in leukaemia

The influence of iron metabolism, both on the invading bacterial pathogen and in the host is widespread and often appears to be crucial in determining the outcome of an infection. This study involved the investigation of leukaemia, a clinical disease where abnormal availability of iron may play a part in predisposing patients to bacterial infection. The iron status throughout a Gram-negative septicaemia and in 20 random, newly diagnosed leukaemic patients was assessed. The results revealed that the majority of the patients exhibited high serum iron levels and serum transferrin saturation often at 100%, with an inability to reduce the latter to within normal values during an infection episode. The antibody response to P.aeruginosa, E.coli and K.pneumoniae outer membrane protein (OMP) antigens were investigated by immunoblotting with sequential serum samples during infection in the leukaemic host. Antibodies to all the major OMPs, were observed, although recognition of iron-regulated membrane proteins (IRMPs) was in many cases weak. Results from the enzyme-linked immunosorbent assay indicated that in all patients antibody titre in response to infection was poor. Sub-MICs of mitomycin C significantly altered the surface characteristics of P.aeruginosa. The silver-stained SDS-PAGE gels of proteinase K digested whole cell lysates of strains PAO1, 6750, M7 and PAJ indicated that core LPS was affected in the presence of mitomycin C. In contrast, the rough strain AK1012 showed no observable differences. Results obtained using quantitative gas-liquid chromatographic analysis showed the amount of LPS fatty acids to be unaffected, however, the KDO and carbohydrate content in strains PAO1, 6750 and M7 under Fe+ and Fe- growth conditions were decreased by up to 4-fold in the presence of mitomycin C, indicating perturbed expression of LPS. The cell surface became significantly more hydrophobic in the P.aeruginosa strains, except AK1012 which was comparatively unaffected. The induction of protein G (OprG) in P.aeruginosa was found to be a sensitive indicator of media iron. The data indicated that expression of OprG can be modulated by growth rate/phase, availability of iron and by the presence of ciprofloxacin in the growth medium.

An advanced thyristor valve for HVDC transmission

The development of an advanced outdoor valve requires coordinated research in the areas of light-triggered self-protecting thyristors, light triggering systems, insulation, cooling and mechanical design aspects. This thesis addresses the first two areas primarily, with a conceptual discussion of the remainder. Using the experience gained from evaluation of a prototype thyristor and computer IKdelling of turn-on behaviour, a light-triggered thyristor with immunity to damage from weak optical triggering and dv/dt triggering was designed, manufactured and evaluated. The optical turn-on process was investigated by measuring currents and voltages in the gate structure during turn-on, and this yielded insights not obtained through conventional measurement techniques. The mechanism by which the thyristor was immune to weak triggering damage is explained, and techniques for optimising the design of the gate structure are proposed. The most significant achievement, however, was the first demonstration of the feasibility of self-protection against forward recovery failure onditions. Furthermore, this was achieved without the need for complex structures or high levels of irradiation. The perfomance of the devices was limited by the inrush capability of the Zones, but it is believed that this can be improved by conventional means. A light triggering system was developed using sem~conductor lasers, and this incorporated several improvements over prior art In terms of optical performance and flexibility.

Development of a multicellular co-culture model of normal and cystic fibrosis human airways in vitro

Cystic fibrosis (CF) is the most common lethal inherited disease among Caucasians and arises due to mutations in a chloride channel, called cystic fibrosis transmembrane conductance regulator. A hallmark of this disease is the chronic bacterial infection of the airways, which is usually, associated with pathogens such as Pseudomonas aeruginosa, S. aureus and recently becoming more prominent, B. cepacia. The excessive inflammatory response, which leads to irreversible lung damage, will in the long term lead to mortality of the patient at around the age of 40 years. Understanding the pathogenesis of CF currently relies on animal models, such as those employing genetically-modified mice, and on single cell culture models, which are grown either as polarised or non-polarised epithelium in vitro. Whilst these approaches partially enable the study of disease progression in CF, both types of models have inherent limitations. The overall aim of this thesis was to establish a multicellular co-culture model of normal and CF human airways in vitro, which helps to partially overcome these limitations and permits analysis of cell-to-cell communication in the airways. These models could then be used to examine the co-ordinated response of the airways to infection with relevant pathogens in order to validate this approach over animals/single cell models. Therefore epithelial cell lines of non-CF and CF background were employed in a co-culture model together with human pulmonary fibroblasts. Co-cultures were grown on collagen-coated permeable supports at air-liquid interface to promote epithelial cell differentiation. The models were characterised and essential features for investigating CF infections and inflammatory responses were investigated and analysed. A pseudostratified like epithelial cell layer was established at air liquid interface (ALI) of mono-and co-cultures and cell layer integrity was verified by tight junction (TJ) staining and transepithelial resistance measurements (TER). Mono- and co-cultures were also found to secrete the airway mucin MUC5AC. Influence of bacterial infections was found to be most challenging when intact S. aureus, B. cepacia and P. aeruginosa were used. CF mono- and co-cultures were found to mimic the hyperinflammatory state found in CF, which was confirmed by analysing IL-8 secretions of these models. These co-culture models will help to elucidate the role fibroblasts play in the inflammatory response to bacteria and will provide a useful testing platform to further investigate the dysregulated airway responses seen in CF.

Grating Deices in polymer optical fibre

This thesis presents the fabrication of fibre Bragg gratings (FBGs) and long period gratings (LPGs) in polymer optical fibre (POF). Possible fabrication techniques were discussed to fabricate FBGs in polymer optical fibre including a detailed description of the phase mask inscription technique used to fabricate FBGs in both single and multi mode microstructured polymer optical fibre (mPOF). Complementing the fabrication of polymer optical fibre Bragg gratings (POFBGs), a technique has been developed to permanently splice POF to silica optical fibre with the use of an optical adhesive. This allowed for the fabricated POFBGs to be characterised away from the optical table, allowing for application specific characterisation. Furthermore Bragg gratings have been fabricated in polymer POF with a Bragg response within the 800nm spectral region. Within this spectral region, POF predominantly manufactured from PMMA experiences considerably smaller attenuation losses when compared to the attenuation losses within the 1550nm spectral region. The effect of thermally annealing fabricated POFBGs has been studied. This included demonstrating the ability to tune the Bragg wavelength of a POFBG sensor to a desired wavelength. Thermal annealing has also been used to manufacture wavelength division multiplexed sensors with the use of a single phase mask. Finally POFBGs have been fabricated in Topas Cyclic Olefin Copolymer. Fabrication of Bragg gratings within this copolymer allowed for the first demonstration of near immunity to relative humidity whilst monitoring changes in temperature of the environment the POFBG sensor was in. Bragg gratings fabricated in the Topas copolymer demonstrated sensitivity to relative humidity which was 65 times less than that of a PMMA based POFBG sensor. This decrease in sensitivity has the potential to significantly reduce the potential of cross sensitivity to relative humidity whilst being employed to monitor measurands such as temperature and axial strain.

Dealing with cell death – an ageing problem?

The aged population have an increased susceptibility to infection, therefore function of the innate immune system may be impaired as we age. Macrophages, and their precursors monocytes, play an important role in host defence in the form of phagocytosis, and also link the innate and adaptive immune system via antigen presentation. Classically-activated ‘M1’ macrophages are pro-inflammatory, which can be induced by encountering pathogenic material or pro-inflammatory mediators. Alternatively activated ‘M2’ macrophages have a largely reparative role, including clearance of apoptotic bodies and debris from tissues. Despite some innate immune receptors being implicated in the clearance of apoptotic cells, the process has been observed to have a dominant anti-inflammatory phenotype with cytokines such as IL-10 and TGF-ß being implicated. The atherosclerotic plaque contains recruited monocytes and macrophages, and is a highly inflammatory environment despite high levels of apoptosis. At these sites, monocytes differentiate into macrophages and gorge on lipoproteins, resulting in formation of ‘foam cells’ which then undergo apoptosis, recruiting further monocytes. This project seeks to understand why, given high levels of apoptosis, the plaque is a pro-inflammatory environment. This phenomenon may be the result of the aged environment or an inability of foam cells to elicit an anti-inflammatory effect in response to dying cells. Here we demonstrate that lipoprotein treatment of macrophages in culture results in reduced capacity to clear apoptotic cells. The capability of lipoprotein treated macrophages to respond to inflammatory stimuli is also shown. Monocyte recruitment to the plaque is currently under study, as is apoptotic cell-mediated immune modulation of human monocyte-derived macrophages.

Does metabolic reprogramming underpin age-associated changes in T cell phenotype and function?

T cells are required for an effective adaptive immune response. The principal function of T cells is to promote efficient removal of foreign material by identifying and mounting a specific response to nonself. A decline in T cell function in aging is thought to contribute to reduced response to infection and vaccination and an increase in autoimmunity. This may in part be due to the age-related decrease in naïve CD4+ T cells and increase in antigen-experienced CD4+ T cells, loss of redox homeostasis, and impaired metabolic switching. Switching between subsets is triggered by the integration of extracellular signals sensed through surface receptors and the activation of discrete intracellular metabolic pathways. This article explores how metabolic programming and loss of redox homeostasis during aging may contribute to age-associated changes in T cell phenotype and function. © 2014 Elsevier Inc.

Phosphatidylserine-containing liposomes:modulators of rhinovirus induced inflammatory responses

Background: Human rhinoviral infections are major contributors to the healthcare burden associated with acute exacerbations of asthma. We, and others have recently demonstrated that rhinovirus (RV)-induced inflammatory responses are mediated by multiple signalling mechanisms, such as IL-1/MyD88 (1) and TLR3/RIGI (2). We have also previously published work showing that TLR signalling is effectively inhibited by phosphatidylserine-containing liposomes (SAPS), through the disruption of membrane microdomains (3). Evidence has also suggested that membrane microdomains may influence infections with RV. In this study, we explored the ability of SAPS to modulate responses to the natural viral pathogens, RV-1B and RV-16. Method: The immortalized bronchial epithelial cell line, BEAS-2B or primary bronchial epithelial cells were infected with RV-1B or RV-16 at a TCID50/ml of 19107 for 1 h. Immediately following infection, various concentrations of SAPS were added and changes in cytokine release were measured at 24 h. SAPS remained present throughout. Type I and III interferon (IFN) expression and rates of viral replication were measured by quantitative PCR. Virus quantification was also performed using a viral CPE assay, and IFN signalling was measured by western blot. Liposome stability was characterised and intracellular trafficking of fluorescently labelled SAPS in BEAS-2B cells was investigated using confocal microscopy. For in vivo studies, female wt Balb/c mice were pre-treated with SAPS for 2 h prior to infection with RV as previously described and changes in BAL cell number, BAL cytokine production and viral replication were quantified (4). Results: Characterisation of SAPS liposomes by mass spectrometry showed no obvious signs of oxidation over the time period tested, and liposome size remained constant. Preliminary confocal studies revealed that SAPS was rapidly internalised within the cell and was found to associate with intracellular compartments such as the early endosome and golgi. Viral infected BEAS-2B cells co-incubated with SAPS, showed notably impaired responses to RV as assessed by release of CXCL8 and CCL5. SAPS also reduced RV-induced IFNb production and STAT-1 phosphorylation, without significantly influencing viral replication rates. Modest increases in viral particle production were only observed at 48 and 72 h time points. Suppression of viral-induced cytokine production was also observed in primary bronchial epithelial cells and pilot in vivo studies showed that SAPS results in reduced KC production at 24 h post viral infection, and this was associated with reduced neutrophil numbers within the BAL fluid. Conclusion: Our data demonstrates a potential means of modulating inflammatory responses induced by human rhinovirus.

Trajetória de pessoas com AIDS em situação de vulnerabilidade social: à luz da história oral de vida

The epidemic caused by HIV presents a global, dynamic and unstable phenomenon, which depends on the individual and collective human behavior. Efforts to deconstruct the stigmatized image caused by infection of AIDS are still often associated with adoption of socially unacceptable behavior to be a circumscribed the susceptibilities of vulnerable individuals and communities to infection, illness and death by HIV. This study aimed to: narrate the trajectory of life of people with AIDS more vulnerable enrolled in the Municipal Social Assistance Parnamirim / RN. It is a study of qualitative, exploratory and descriptive approach, taking oral history of life as technical and methodological framework. The colony consisted of 186 people with AIDS. The network was comprised of 13 employees of both sexes, aged between 19 and 62 years old with positive diagnosis and agreed to voluntarily participate. After approval by the Ethics Committee of the Federal University of Rio Grande do Norte (CEP / UFRN), in the opinion No. 719,926 CAAE: 30408114.5.0000.5537 on 6 June 2014 data were collected from August to September 2014. The employees signed the Informed Consent and Informed and letter of assignment. Held transcribing the interviews and later returned to respondents to retest, ie so that they confer what allowed us to carry out transcreation after consecutive readings. The reports were analyzed through Bardin content analysis. Guiding the analysis of the accounts of employees, we find three themes: Prejudice and discrimination in living with AIDS; Reacting to the diagnosis and the accession process to antiretroviral treatment; and religious coping in people with AIDS. It can be concluded in this study, that employees have shown great emotional impact after positive diagnosis for HIV / AIDS, especially with regard to social life, the family ties, work and above all to the prejudice of society. Treatment with antiretroviral drugs was seen as a motivation to regain dreams and plans for a future once uncertain, and even if it is not a cure therapy, provided the employees improved quality of life.

Trajetória de pessoas com AIDS em situação de vulnerabilidade social: à luz da história oral de vida

The epidemic caused by HIV presents a global, dynamic and unstable phenomenon, which depends on the individual and collective human behavior. Efforts to deconstruct the stigmatized image caused by infection of AIDS are still often associated with adoption of socially unacceptable behavior to be a circumscribed the susceptibilities of vulnerable individuals and communities to infection, illness and death by HIV. This study aimed to: narrate the trajectory of life of people with AIDS more vulnerable enrolled in the Municipal Social Assistance Parnamirim / RN. It is a study of qualitative, exploratory and descriptive approach, taking oral history of life as technical and methodological framework. The colony consisted of 186 people with AIDS. The network was comprised of 13 employees of both sexes, aged between 19 and 62 years old with positive diagnosis and agreed to voluntarily participate. After approval by the Ethics Committee of the Federal University of Rio Grande do Norte (CEP / UFRN), in the opinion No. 719,926 CAAE: 30408114.5.0000.5537 on 6 June 2014 data were collected from August to September 2014. The employees signed the Informed Consent and Informed and letter of assignment. Held transcribing the interviews and later returned to respondents to retest, ie so that they confer what allowed us to carry out transcreation after consecutive readings. The reports were analyzed through Bardin content analysis. Guiding the analysis of the accounts of employees, we find three themes: Prejudice and discrimination in living with AIDS; Reacting to the diagnosis and the accession process to antiretroviral treatment; and religious coping in people with AIDS. It can be concluded in this study, that employees have shown great emotional impact after positive diagnosis for HIV / AIDS, especially with regard to social life, the family ties, work and above all to the prejudice of society. Treatment with antiretroviral drugs was seen as a motivation to regain dreams and plans for a future once uncertain, and even if it is not a cure therapy, provided the employees improved quality of life.

Effects of seasonally varying temperature and salinity on the dynamics of sea lice (Lepeophtheirus salmonis)

Sea lice (Lepeophtheirus salmonis) are an economically significant parasite in salmonid aquaculture. They exhibit temperature-dependent development rates and salinity-dependent mortality, which can greatly impact sea lice population dynamics, but no deterministic models have incorporated these seasonal variables. To understand how seasonality affects sea lice population dynamics, I derive a delay differential equation model with temperature and salinity dependence. I find that peak reproductive output in Newfoundland and British Columbia differs by four months. A sensitivity analysis shows sea lice abundance is most sensitive to variation in mean annual water temperature and salinity, whereas it is lease sensitive to infection rate. Additionally, I investigate the effects of production cycle timing on sea lice management and find that optimal production cycle start times are between the 281st and 337th days of the year in Newfoundland. I also demonstrate that adjusting follow-up treatment timing in response to temperature can improve treatment regimes. My results suggest that effective sea lice management requires consideration of local temperature and salinity patterns.

Mecanismos protetores induzidos pelo tratamento prévio com antígeno solúvel de taquizoítas (STAg) no intestino delgado de camundongos BALB/c e C57BL/6 infectados oralmente por Toxoplasma gondii

T. gondii can infect the gut mucosa by direct invasion of epithelial cells in the small intestine and these cells may respond directly to infection promoting a local immune response. C57BL/6 mice orally infected with a high parasitic load of T.gondii are highly susceptible, presenting a lethal ileitis. Recently, it was demonstrated that pretreatment with STAg protects C57BL/6 mice against intestinal pathology in oral T. gondii infection. To investigate the mechanisms induced by STAg in the small intestine in oral T.gondii infection, BALB/c and C57BL/6 mice were treated with STAg 48 hours before oral infection with 30 ME-49 cysts and sacrificed at 8 days of infection. Previous treatment with STAg were able of decrease parasitism and pathology in peripheral organs of BALB/c and C57BL/6 mice and induced a increase in amounts of goblet cells, IgA positive cells, Paneth cells and expression of cryptidin in the small intestine of both lineages of mice, moreover BALB/c mice presented higher amount of these cells comparing with C57BL/6 mice. The results suggests that STAg is able of promoting protective mechanisms in both lineages of mice, although these protection is more evidenced in BALB/c mice, and these mechanisms could be in part mediated by increase in goblet, Paneth and local secretion of IgA in the small intestine of mice orally infected with T.gondii.

An investigation of the molecular interactions between statins and bacterial pathogens, and their combined impact on the human immune system

Statins are a class of drug that inhibits cholesterol biosynthesis, and are used to treat patients with high serum cholesterol levels. They exert this function by competitively binding to the enzyme 3-hydroxy-3-methylglutaryl-CoenzymeA reductase (HMGR), which catalyses the formation of mevalonate, a rate-limiting step in cholesterol biosynthesis. In addition, statins have what are called “pleiotropic effects”, which include the reduction of inflammation, immunomodulation, and antimicrobial effects. Statins can also improve survival of patients with sepsis and pneumonia. Cystic fibrosis (CF) is the most common recessive inherited disease in the Caucasian population, which is characterised by factors including, but not limited to, excessive lung inflammation and increased susceptibility to infection. Therefore, the overall objective of this study was to examine the effects of statins on CFassociated bacterial pathogens and the host response. In this work, the prevalence of HMGR was examined in respiratory pathogens, and several CF-associated pathogens were found to possess homologues of this enzyme. HMGR homology was analysed in Staphylococcus aureus, Burkholderia cenocepacia and Streptococcus pneumoniae, and the HMGR of B. cenocepacia was found to have significant conservation to that of Pseudomonas mevalonii, which is the most widely-characterised bacterial HMGR. However, in silico analysis revealed that, unlike S. aureus and S. pneumoniae, B. cenocepacia did not possess homologues of other mevalonate pathway proteins, and that the HMGR of B. cenocepacia appeared to be involved in an alternative metabolic pathway. The effect of simvastatin was subsequently tested on the growth and virulence of S. aureus, B. cenocepacia and S. pneumoniae. Simvastatin inhibited the growth of all 3 species in a dose-dependent manner. In addition, statin treatment also attenuated biofilm formation of all 3 species, and reduced in vitro motility of S. aureus. Interestingly, simvastatin also increased the potency of the aminoglycoside antibiotic gentamicin against B. cenocepacia. The impact of statins was subsequently tested on the predominant CF-associated pathogen Pseudomonas aeruginosa, which does not possess a HMGR homologue. Mevastatin, lovastatin and simvastatin did not influence the growth of this species. However, sub-inhibitory statin concentrations reduced the swarming motility and biofilm formation of P. aeruginosa. The influence of statins was also examined on Type 3 toxin secretion, quorum sensing and chemotaxis, and no statin effect was observed on any of these phenotypes. Statins did not appear to have a characteristic effect on the P. aeruginosa transcriptome. However, a mutant library screen revealed that the effect of statins on P. aeruginosa biofilm was mediated through the PvrR regulator and the Cup fimbrial biosynthesis genes. Furthermore, proteomic analysis demonstrated that 6 proteins were reproducibly induced by simvastatin in the P. aeruginosa swarming cells. The effect of statins on the regulation of the host-P. aeruginosa immune response was also investigated. Statin treatment increased expression of the pro-inflammatory cytokines IL-8 and CCL20 in lung epithelial cells, but did not attenuate P. aeruginosa-mediated inflammatory gene induction. In fact, simvastatin and P. aeruginosa caused a synergistic effect on CCL20 expression. The expression of the transcriptional regulators KLF2 and KLF6 was also increased by statins and P. aeruginosa, with the induction of KLF6 by simvastatin proving to be a novel effect. Interestingly, both statins and P. aeruginosa were capable of inducing alternative splicing of KLF6. P. aeruginosa was found to induce KLF6 alternative splicing by way of the type 3 secreted toxin ExoS. In addition, a mechanistic role was elucidated for KLF6 in the lung, as it was determined that statin-mediated induction of this protein was responsible for the induction of the host response genes CCL20 and iNOS. Moreover, statin treatment caused a slight increase in infection-related cytotoxicity, and increased bacterial adhesion to cells. Taken together, these data demonstrate that statins can reduce the virulence of CFassociated bacterial pathogens and alter host response effectors. Furthermore, novel statin effectors were identified in both bacterial and host cells.