Thesis (M.S.)--University of Wisconsin--Madison, 1981. / Typescript. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references (leaves 90-92).

Cigarette smoke displays both immunoactivating and immunosuppressive activities. Although the health risks of cigarette smoke are widely reported, the impact and mechanism of cigarette smoke exposure on inflammatory diseases are still unclear. Innate immune responses could be activated shortly after influenza A virus infection and then trigger inflammation in the lung. Until now, the effect of prior cigarette smoke on the inflammation caused by the following influenza A virus infection has not been fully understood and there is no study about the role of cigarette smoke in 2009 pandemic H1N1 (pdmH1N1) and avian H9N2 (H9N2/G1) infection. In this study, C57/B6N mice were whole body exposed to 4% cigarette smoke 4 hours per day for 21 days and then infected with pdmH1N1 or H9N2 virus. Some mice were exposed to room air in parallel as the control. Nicotine was also used to mimic the effect of the cigarette smoke. Human lung adenocarcinoma epithelial cell line A549 or human monocyte-derived macrophages were pre-treated with 10μM nicotine, and then infected with pdmH1N1 or H9N2 virus. The expression of cytokines and chemokines was examined in the supernatant. C57/B6N mice were treated with 24mg/kg/day nicotine or sterile water subcutaneously for 21 days and then infected with pdmH1N1 or H9N2 virus. Survival, body weight changes, lung viral loads, inflammatory responses and immune cells that infiltrated into the lung were analyzed. Cigarette smoke exposure alone significantly up-regulated the lung inflammation, confirmed by the dramatically decreased body weight gain and increased inflammatory response compared with the control mice. Such prior cigarette smoke exposure significantly reduced the disease severity induced by subsequent pdmH1N1 or H9N2 virus infection. For pdmH1N1 infection, cigarette smoke exposure group had a significantly lower mortality than the control group, which might be due to the lower inflammatory response at day 5 post virus infection. Similarly, after H9N2 virus infection, the mice in cigarette smoke exposure group displayed a significantly milder disease, as evidenced by less weight loss, weaker inflammatory response as well as lower number of immune cells infiltrating into the lung compared to the control group. There was no difference for the lung viral loads between the two groups upon both pdmH1N1 and H9N2 virus infection. Besides, pre-treating the A549 and primary human macrophages with nicotine decreased the expression of some cytokines and chemokines after pdmH1N1 or H9N2 virus infection. The mice treated with nicotine displayed significantly less weight loss and lower inflammatory response than control mice treated with sterile water upon pdmH1N1 or H9N2 virus infection. Collectively, our study demonstrated that the immunosuppressive effect of cigarette smoke was responsible for amelioration of pdmH1N1 and H9N2 pathogenicity by dampening the hyper-reaction of inflammatory response without any direct effect on viral replication. Nicotine, an anti-inflammatory factor, was the key component in cigarette smoke that was responsible for this immunosuppressive effect. Our study provided the first in vivo evidence that cigarette smoke, mostly due to nicotine, could protect against the pathogenicity of not only pdmH1N1, but also H9N2 virus. / published_or_final_version / Paediatrics and Adolescent Medicine / Doctoral / Doctor of Philosophy

No description available.

Chronic obstructive pulmonary disease (COPD) is a highly prevalent disease in the elderly. It is currently the fourth leading cause of death and will become the third by 2030. Cigarette smoke is the major cause of COPD pathogenesis, resulting from the burden of oxidants, which stimulates the production of inflammatory chemokines, leading to the influx of inflammatory cells into the airways and causing chronic inflammation. Due to lung infection by bacteria, such as Pseudomonas Aeruginosa during acute exacerbation in COPD, cigarette smoking might induce an immunosuppressive effect, which leads to bacteria colonization in the airways and further contributes to the chronic inflammation in the airway of COPD. Furthermore, cigarette smoke-induced production of reactive oxygen species (ROS) also plays an important role in the pathogenesis of COPD, however, N-acetyl-L-cysteine (NAC), which has been administered for the treatment of COPD as a mucolytic agent, also showed antioxidant and anti-inflammatory effect. The exact mechanism or cellular pathway through which cigarette smoke suppresses bacteria-induced inflammatory response and how NAC acts as an anti-inflammatory agent still remains uncertain. This study aims to investigate the effect of cigarette smoke and lipopolysaccharide (LPS) alone or in combination on the release of pro-inflammatory chemokines and to elucidate cigarette smoke-induced chemokines release in the presence and absence of NAC. Both cigarette smoke and LPS alone induced the release of interleukin-8 (IL-8) and monocyte chemoattractant protein-1 (MCP-1).Cigarette smoke suppressed the LPS-induced IL-8 and MCP-1release. NAC reduced both basal and cigarette smoke-induced secretion of these inflammatory chemokines. Moreover, Western blot demonstrated that cigarette smoke activated AMPKα phosphorylation, which was suppressed with NAC pretreatment, suggesting that NAC might have inhibitory effect on the release of chemokine release via the AMPK pathway. Our current data suggests that there may be a link between ROS generation to AMPK activation and chemokine release in BEAS-2B cells. / published_or_final_version / Pharmacology and Pharmacy / Master / Master of Medical Sciences

Cigarette smoke is the leading cause of preventable mortality worldwide. This excess death is attributable to an increased risk of acquiring a variety of conditions, including chronic respiratory/cardiovascular diseases and various types of cancer. Smokers are additionally predisposed to develop infectious diseases, notably including pneumonia caused by the influenza virus, one of the most prevalent and burdensome pathogens in existence today. Although cigarette smoke is well known to modulate many aspects of the immune system, the specific mechanisms by which this predisposition is mediated are incompletely understood. Also unclear is the effect of cigarette smoke on responses to intranasal immunization strategies aimed at eliciting immunity against pathogens such as influenza in the upper airways, where protection may substantially contribute to sterilizing immunity. This PhD thesis focused primarily on addressing these knowledge gaps. In the first study, we assessed the effect of cigarette smoke on antibody induction following intranasal immunization in the upper airways of mice, finding that smoke exposure attenuated antigen-specific IgA induction in the upper respiratory tract, reproductive tract, and systemic circulation. In addition, we found that these nasal IgA demonstrated a reduced antigen-binding avidity in the acute post-immunization period. Mechanistically, deficits in nasal IgA were associated with a reduced accumulation of antigen-specific IgA antibody-secreting cells (ASCs) in the nasal mucosa, induction of these cells in nasal-draining lymphoid tissues, and upregulation of molecules critical to ASC homing (vascular cell adhesion molecule-1; VCAM-1) and IgA transepithelial transport (polymeric immunoglobulin receptor; pIgR) in the nasal mucosa. Ultimately, in tandem with recent clinical work published by others, our study strongly suggests that cigarette smoke can attenuate IgA induction in the upper airways, which may have implications for aspects of intranasal vaccine efficacy. Thus, smoking status should be more consistently considered in the design of clinical trials for IgA-oriented intranasal vaccines. The second study did not assess smoking and host defense directly, but rather served to optimize protocols for assessing immunoglobulins in human mucoid respiratory samples as a precursor to future studies in smoking-related disease. In this regard we found that, relative to phosphate-buffered saline (PBS), dithiothreitol (DTT)-based processing of human sputum samples increased total IgA yields, decreased IgE yield, and improved the detection of a specific IgG autoantibody. These findings suggest that processing choices for human mucoid respiratory samples should be made with specific goals in mind as they pertain to antibody isotype(s) of interest. Finally, in the third study we investigated potential mechanisms by which cigarette smoke exposure promotes influenza, given that smokers are at increased risk of acquiring the pathogen, progressing to severe disease, and being admitted to hospital/ICU following infection. In doing so, we found that concurrent smoke exposure increased morbidity, hypoxemia, pulmonary edema, neutrophilia, and ultimately mortality in a mouse model of H1N1 infection. These changes were associated with an increased accumulation of viral (v)RNA in cells independent of any change in the shedding of replication-competent viral particles. Using a novel dysregulation score approach, we found that interleukin (IL)-6 and colony-stimulating factor (CSF)3 expression was highly exacerbated in the lungs and circulation of smoke-exposed, infected mice relative to controls. Supplementation of recombinant (r)CSF3 increased morbidity, hypothermia and edema, while blockade of the cognate receptor (CSF3R) improved alveolar-capillary barrier function. On the cellular level, single cell RNA-sequencing revealed a shift in the distribution of Csf3+ cells towards neutrophils. Finally, deep transcriptional analysis of neutrophils revealed a gene signature that was largely indicative of an exacerbated form of typical disease with select unique regulatory elements. Ultimately, this work identifies potential therapeutic targets (CSF3R signaling, excess vRNA accumulation) for the treatment of cigarette smoke-augmented influenza, and warns against clinical rCSF3 therapy to treat neutropenia during viral infectious disease. In conclusion, the work presented in this PhD dissertation expands our understanding of the relationship between cigarette smoke and antimicrobial host defense as it pertains to both IgA immunity in the upper airways, and the pathogenesis of cigarette smoke-augmented influenza. / Thesis / Doctor of Philosophy (PhD) / Cigarette smoke exposure is well known to have many harmful effects on human health, including through its ability to promote various infectious diseases such as influenza. However, the mechanisms by which it promotes infection are not fully known. This is an important knowledge gap given that over 1.1 billion individuals continue to smoke worldwide, and a large number of people are exposed to the harmful effects of second-hand smoke, both with fatal consequence. The central goal of this thesis was to gain a better understanding of this relationship between cigarette smoke and infectious disease, specifically by assessing how smoke exposure impacts immune responses in the upper and lower airways. In the first study, we found that smoke exposure interferes with the ability to activate immunoglobulin (Ig)A antibody responses in the nasal passages of mice, which may have important implications for human nasal vaccination strategies. The second study investigated different methods with which to best measure antibodies in human respiratory samples. Finally, in the third study we defined a role for a specific molecule, CSF3, in worsening health in a mouse model of concurrent cigarette smoke and influenza infection. Overall, this work provides new insights into the ways in which smoking can increase the risk of respiratory infection, thereby informing the future design and testing of vaccines and treatments for use in our highly smoke-exposed global population.

Thesis (Ph. D.)--Chemistry and Biochemistry, Georgia Institute of Technology, 2005. Directed by Boris Mizaikoff. / Hunt, William, Committee Member ; Weck, Marcus, Committee Member ; Mizaikoff, Boris, Committee Chair ; Janata, Jiri, Committee Member ; Orlando, Thomas, Committee Member. Includes bibliographical references.

Cigarette smoking (CS) represents the main causative agent underlying development and progress of COPD. Recently, involvement of CS in the pathogenesis of COPDassociated muscle abnormalities is becoming increasingly evident. Nevertheless, involved triggers and underlying mechanisms remain largely unknown. This study was conceived in order to examine effects of cigarette smoke exposure on skeletal muscle morphology, vascular supply and function. For this purpose, we have specifically designed murine COPD/emphysema model and gastrocnemius muscle was examined, while in vitro experiments were conducted using murine C2C12 skeletal muscle myocytes. In addition to the mild emphysematous changes present in the lungs of CS-exposed mice, our results demonstrated evident signs of muscle atrophy reflected by decreased fiber cross-sectional area, profound fiber size variation and reduced body mass. Furthermore, we have observed impairment in terminal myogenesis and lower number of myonuclei in skeletal muscles of CS-exposed animals despite evident activation of muscle repair process. Additionally, our results demonstrate capillary rarefaction in skeletal muscles of CS-exposed animals which was associated with deregulation of hypoxia-angiogenesis signaling, reduced levels of angiogenic factors such as HIF1-α and VEGF and enhanced expression of VHL and its partner proteins PHD2 and Ube2D1. The results of our in-vitro experiments demonstrated that VHL and its ubiquitination machinery can be synergistically regulated by TNF and hypoxia consequentially impairing angiogenic potential of skeletal muscle myocytes. Finally, we have shown that CS elicits chronic ER stress in murine skeletal muscles which is associated with activation of ERAD and apoptotic pathways as mirrored by elevated expression of Usp19, caspase 12 and caspase 3 in skeletal muscles of CSexposed animals. Moreover, molecular and morphological alterations in CS-exposed mice resulted in impairment of muscle function as reflected by their impaired exercise capacity. Taken together, from our results it is evident that cigarette smoke exposure elicits set of morphological, vascular and functional changes highly resembling those observed in COPD. Additionally, CS induces wide range of molecular alterations and signaling pathway deregulations suggesting profound effects of cigarette smoke exposure on skeletal muscle cell homeostasis.

Streptococcus pneumoniae is a common human pathogen, causing severe and often life-threatening respiratory tract infections. Even though most patients receive appropriate antimicrobial chemotherapy, a significant percentage still die. Cigarette smoking is a well-documented risk factor for severe pneumococcal pneumonia; however, the microbiological/ immunological mechanisms which predispose smokers to infection are not yet completely understood. The pneumococcal toxin, pneumolysin, is a major virulence factor of S. pneumoniae; it forms pores in eukaryotic cell membranes, resulting in the influx of extracellular Ca2+. Biofilm is a self-generated polymer matrix, used by microbial pathogens to isolate themselves from both host defences, as well as antibiotics. In this dissertation, the effects of cigarette smoke condensate (CSC) on the bioactivity of pneumolysin, as well as on the growth of, and production of biofilm by Streptococcus pneumoniae are described. A clinical isolate of S. pneumoniae, strain 172, was used for both growth and biofilm determinations in the presence or absence of concentrations of CSC (20-160 μg/ml), representative of the smoking habit. Growth and biofilm determinations were performed using spectrophotometric procedures, viability by standard colony forming unit procedures, and the effect of the condensate on pneumolysin bioactivity using a fura-2/AM-based spectrofluorometric procedure. Exposure of the pneumococcus to CSC resulted in a dose-dependent increase in biofilm formation which achieved statistical significance (P≤0.05) at concentrations of 80 and 160 μg/ml, in the setting of modest effects on bacterial growth. These findings were not unique to S. pneumoniae since exposure of Staphylococcus aureus, a known biofilm former, to CSC showed similar results. Exposure of pneumolysin to CSC (20 and 40 μg/ml) was accompanied by attenuation of the biological activity of the toxin, resulting in impaired pore-forming ability manifest as a considerable reduction in influx of extracellular Ca2+ following exposure of isolated neutrophils to the toxin. It is possible that CSC acts as a stressor to the bacteria, thereby enhancing biofilm formation and consequently persistence in the respiratory tract. These effects of the toxin may be complemented by inactivation of pneumolysin, presumably by pro-oxidative mechanisms, affecting innate cellular host defences. These mechanisms may underpin the predisposition of smokers to develop severe pneumococcal infections. / AFRIKAANS : Streptococcus pneumoniae is ‘n algemene menslike patogeen, wat ernstige en soms lewensbedreigende lugweg infeksies veroorsaak. Alhoewel die meeste pasiënte geskikte antimikrobiese chemoterapie ontvang, sterf ‘n betekenisvolle hoeveelheid steeds. Sigaretrook is ‘n welbekende risikofaktor vir ernstige pneumokokkale pneumonie; die mikrobiologiese/ immunologiese meganisme wat rokers vatbaar maak vir infeksie word egter nog nie heeltemal verstaan nie. Die pneumokokkale toksien, pneumolisien, is ‘n belangrike virulensie faktor van S. pneumoniae; dit vorm porieë in die eukariotiese selmembrane wat ‘n invloei van ekstrasellulêre Ca2+ tot gevolg het. Biofilm is ‘n self-genererende polimeermatriks, wat deur mikrobiese patogene gebruik word om hulself teen gasheerverdedigings meganismes en antibiotika te isoleer. In hierdie verhandeling word die uitwerking van sigaretrook kondensaat (SRK) op die bioaktiwiteit van pneumolisien, asook die effekte op groei en die produksie van biofilm deur S. pneumoniae, beskryf. ‘n Kliniese isolaat van S. pneumoniae, stam 172, is gebruik vir beide groei en biofilm bepalings, in die teenwoordigheid en afwesigheid van SRK (20-160 μg/ml), wat verteenwoordigend van die rook gewoonte is. Groei en biofilm bepalings is uitgevoer deur gebruik te maak van spektrofotometriese prosedures, lewensvatbaarheid, deur standard kolonievormende eenheid prosedures, en die effek van die kondensaat op die pneumolisien bioaktiwiteit, deur fura-2/AM-gebaseerde spektrofluorometriese prosedures. Blootstelling van die pneumokokkus aan SRK het in ‘n dosis-verwante verhoging in biofilm-formasie tot gevolg gehad, wat statistiese betekenisvolheid (P≤0.05) bereik het teen konsentrasies van 80 en 160 μg/ml, teenoor die agtergrond van beskeie effekte op bakteriële groei. Hierdie bevindinge is nie uniek aan S. pneumoniae nie, aangesien blootstelling van Staphylococcus aureus, ‘n bekende biofilm-vormer, aan SRK soortgelyke resultate gelewer het. Blootstelling van pneumolisien aan SRK (20-160 μg/ml) was geassosieer met die verswakking van die biologiese aktiwiteit van die toksien, met verminderde porie-vormende vermoë, wat gemanifesteer het met ‘n ongelooflike verlaging in die invloei van ekstrasellulêre Ca2+ na die blootstelling van geïsoleerde neutrofiele aan die toksien. Dit is moontlik dat die SRK as ‘n stressor vir die bakterieë optree, en daardeur biofilm formasie en gevolglik hardnekkige infeksies in die lugweg tot sellulêre gasheervededing beïnvloed. Hierdie effekte op die toksien mag gekomplimenteer word deur die inaktivering van pneumolisien, heelwaarskynlik deur pro-oksidatiewe meganismes, wat die vatbaarheid van rokers om ernstige pneumokokkale infeksies te ontwikkel, beklemtoon. / Dissertation (MSc)--University of Pretoria, 2011. / Immunology / Unrestricted

Rationale. Cigarette smoking is a well-known risk factor for cardiovascular disease, including arterial diseases such as atherosclerosis and abdominal aortic aneurysm. However, our understanding of how exposure to cigarette smoke impacts arterial disease pathogenesis is not well known. Consequently, this doctoral thesis focuses on understanding the development of atherosclerosis and aortic aneurysm in the context of exposure to cigarette smoke. In particular, since monocytes and macrophage are key immune cells implicated in arterial pathology, this work concentrates on understanding the impact of cigarette smoke exposure on the function and kinetics of monocytes and arterial macrophages. Main Findings. Using a mouse model that combines two clinically relevant risk factors, hyperlipidemia and cigarette smoke, we showed that smoke exposure increases atherosclerosis and induces the spontaneous formation, progression, and rupture of abdominal aneurysms. We also provide experimental evidence that atherosclerosis strongly associates with regions of elastin damage and arterial dilation, suggesting atherogenesis may directly contribute to abdominal aneurysm formation. Given the importance of macrophages in arterial disease, we investigated arterial macrophage heterogeneity and function following exposure to cigarette smoke. We report that cigarette smoke exposure increased the abundance of arterial monocytes and macrophages, whereas heterogeneity was primarily driven by hypercholesterolemia in aneurysmal tissue. Specifically, hypercholesterolemia is associated with an increase in macrophage populations with putative functions in inflammation and tissue remodelling including Trem2 foamy macrophages, inflammatory macrophages, and interferon-inducible macrophages. Moreover, we demonstrated that arterial macrophages play a critical role in elastin fragmentation within the arterial wall of smoke exposed mice. Finally, we investigated the impact of cigarette smoke on kinetic factors that can contribute to arterial macrophage accumulation. We found that, despite increased development of arterial disease, exposure to cigarette smoke is associated with an overall suppression of circulating monocytes and pro-inflammatory cytokines. Using a parabiosis model, we show monocyte recruitment is significantly increased and is likely a key factor contributing to accumulation of arterial macrophages following exposure to cigarette smoke. We also present evidence suggesting that endothelial dysfunction, related to a loss of endothelial nitric oxide synthase, contributes to increased arterial monocyte recruitment following exposure to cigarette smoke. Conclusions and Significance. Overall, we provide evidence that atherosclerosis likely contributes to abdominal aneurysm pathology in a model of cigarette smoke-induced aneurysm formation. We further provide insight into how tobacco smoke promotes arterial disease development through increased local accumulation of arterial macrophages despite suppressed monopoiesis and systemic inflammation. We identify monocyte recruitment and endothelial dysfunction as key factors contributing to the increased accumulation of arterial macrophages, with no overall differences in macrophage heterogeneity, following smoke exposure. In addition to providing insight into the increased risk of arterial disease following exposure to cigarette smoke, this study also provides experimental evidence that atherogenesis can contribute to abdominal aneurysm pathology. Overall, this thesis furthers our understanding of arterial disease pathogenesis and can provide a foundation for further mechanistic or therapeutic focused research aimed at reducing the burden of cardiovascular disease. / Thesis / Doctor of Philosophy (PhD) / Diseases that affect the heart and major blood vessels are one of the leading causes of illness and death worldwide. Atherosclerosis is one such disease caused by the buildup of fatty deposits in the walls of major blood vessels called arteries. This buildup can eventually block the artery and lead to a heart attack or stroke. Abdominal aortic aneurysms are another type of disease that affects arteries. In this case, the walls of the artery grow weak and begin to balloon out until the artery eventually breaks causing severe internal bleeding and death. One of the most important cells involved in the development of atherosclerosis and aneurysms is the macrophage, a type of white blood cell that is an important part of the immune system and found in diseased arteries. Although we know that cigarette smoking is one of the most significant risk factors for developing atherosclerosis and abdominal aneurysms, we do not fully understand why. Therefore, the goal of this thesis project was to investigate how cigarette smoke affects the development of arterial disease with a focus on understanding how it impacts the movement and function of macrophages. Using a mouse model, we found that the development of atherosclerosis and aneurysm are likely related, and also identified ways that exposure to cigarette smoke increases the numbers of macrophages in arteries. This work advances our understanding of how arterial diseases may be related and provides insight into how smoking can increase the risk of developing arterial disease.

Chronic obstructive pulmonary disease (COPD) is a leading cause of global morbidity and mortality, with the potential to afflict as many as half of the 1.1 billion smokers in the world. The inflammatory response to cigarette smoke is believed to mediate the progressive and irreversible loss of lung function that characterizes COPD. The greatest burden of the disease arises from episodes of worsened symptoms and inflammation, usually triggered by microbial infection. Currently, the mechanisms that drive cigarette smoke-induced inflammation are being elucidated but ambiguity remains about this response and the inflammatory response engaged in a smoke-exposed lung experiencing a microbial infection. This thesis sought to investigate inflammatory mediators induced by cigarette smoke and those induced by bacteria, the most common cause of infectious exacerbations of COPD, in the context of smoke exposure. In chapter two we investigated the role of Breast Regression Protein-39 (BRP-39), a gene commonly observed to be increased under inflammatory conditions, in the inflammatory response to cigarette smoke. In order to determine the mechanisms of BRP-39 induction, its expression and inflammation was assessed in IL-13, IL-18, and IL-1R1 deficient mice. BRP-39 was found to be redundant in cigarette smoke-induced inflammation, but these data confirmed that IL-1R1 was a crucial mediator of this response. After examining the inflammatory response elicited by smoke alone, we investigated the importance of IL-1 signaling in a model of bacterial exacerbation of cigarette smoke-induced inflammation. We found that the exacerbated neutrophilia that typifies the response of a smoke-exposed lung to bacteria was dependent on IL-1α-mediated production of the CXCR2 ligand CXCL5. This study identified the unique phenomenon that cigarette smoke primes alveolar macrophages to produce excessive amounts of IL-1α in response to bacterial stimuli. The purpose of the final study of the thesis was to more comprehensively characterize the extent to which cigarette smoke changes the phenotype of macrophages. Examining total gene expression by microarray found that smoke-exposed alveolar macrophages were in a proliferative state expressing a unique profile of inflammatory mediators. Further analysis revealed that this was likely the result of a pulmonary environment rich in growth factors. Taken together, these data provide detail to the understanding of the biological process of inflammation that drives the pathogenesis of COPD. These studies identify a phenomenon that predisposes smokers to experience more severe responses to bacteria and reinforces the targeting of IL-1 signaling in the treatment of COPD. / Thesis / Doctor of Philosophy (Medical Science)