This study is concerned with a critical comparison of the passive transfer mechanisms of tuberculin and chemical contact hypersensitivities in the guinea pig by use of a four phase experimental approach.

Fullerenes are carbon nanospheres that can be solublized by the addition of polar chemical groups to the carbon cage, forming fullerene derivatives. One specifically derivatized fullerene compound, termed C70-Tetragylocolate (C70-TGA), has been shown to stabilize mast cell responses in vitro thus we hypothesized it may have an effect on mast cell-driven diseases such as asthma and systemic anaphylaxis. To observe the effects of C70-TGA on systemic anaphylaxis, mice were subjected to a model of passive systemic anaphylaxis. In this model, mice were injected with DNP-specific IgE 16 hours prior to challenge, then treated with C70-TGA. Immediately prior to DNP challenge, mice were subjected to a second injection of C70-TGA. Following DNP challenge, body temperature was recorded and blood was collected for quantitation of histamine levels. Treatment with C70-TGA significantly reduced body temperature drop associated with systemic anaphylaxis and serum histamine levels. To observe the effects of C70-TGA on chronic features of asthma in vivo, we utilized a heavily MC influenced model of asthma pathogenesis. Mice were sensitized by intraperitoneal (i.p.) injection of ovalbumin (OVA) in saline, challenged intranasally (i.n.) with OVA, and one of two treatment strategies was pursued. In one, C70-TGA was given i.n. throughout disease development. In the other, C70-TGA was given following an initial set of challenges to allow disease to develop prior to treatment; mice were then re-challenged with OVA to assess the effect on established disease. We found that C70-TGA treatment significantly reduced airway inflammation and eosinophilia and dramatically reduced bronchoconstriction in either model. Cytokines IL-4 and IL-5 and serum IgE levels are significantly reduced in C70-TGA treated animals. Interestingly, we also saw an increase in the anti-inflammatory eicosanoid 11, 12-epoxyeicosatreinoic acid (11,12-EET) in the BAL fluid, suggesting the involvement of this mediator in C70-TGA inhibition. Further experiments utilizing an inhibitor of 11,12-EET formation (6-(2-Propargyloxyphenyl)hexanoic acid) and a structural analog of 14,15-EET (14,15-EE-5(Z)-E) in vivo indicate that these mediators are closely associated with C70-TGA mediated inhibition as their inhibition reverses the anti-inflammatory effects of C70-TGA. Importantly, mice did not exhibit any acute toxicity following C70-TGA treatment and liver and kidney function were normal. Collectively, these results show that the fullerene C70 derivative C70-TGA is capable of dampening severe allergic responses including systemic anaphylaxis, airway inflammation, and bronchoconstriction. The mechanism of inhibition is through the upregulation of the anti-inflammatory EETs, which may dampen mast cell degranulation in vivo, thus contributing to the inhibitory effect of C70-TGA on allergic disease.

ADAM10, ADAM17, and SPAG6 contribute significantly to humoral immunity and secondary lymphoid tissue architecture. ADAM10 and ADAM17 are two closely related zinc-metalloproteinases. Through cleavage of their ligands CD23 and TNF, respectively, they greatly influence IgE production and secondary lymphoid tissue architecture maintenance. Th1 prone WT strains initially exhibit increased ADAM17 and TNF yet reduced ADAM10 relative to Th2 prone WT strains. In the absence of B cell ADAM10, a compensatory increase in ADAM17 and TNF cleavage is noted only in Th1 prone C57Bl/6, not Th2 prone Balb/c. B cell TNF homeostasis is important for maintaining secondary lymphoid tissue architecture. We show for the first time that excessive B cell TNF production in C57-ADAM10B-/- lymph nodes contributes to loss of B/T segregation, increased HEV number and size, fibrosis, loss of FDC networks, and impaired germinal center formation. Furthermore, B cell ADAM10, which enhances IgE production through CD23 cleavage, is shown to be a marker of Th2 susceptibility. B cell ADAM10 is elevated in Th2 prone mouse strains and allergic patients compared to Th1 prone controls and as B cell ADAM10 level increases, so does IgE production. Lastly, the B cell profile of allergic patients is determined to be B cell ADAM10highADAM17lowTNFlow. Furthermore, the mechanism underlying reduced class-switched antibody production in C57-ADAM10B-/- mice is explored. C57-ADAM10B-/- B cells exhibit a B10, or IL-10 producing, phenotype, which is linked to reduced antibody production. Furthermore, increased Tregs noted in C57-ADAM10B-/- mice contributed to reduced class switched IgE production and disease parameters following a house dust mite airway inflammation challenge. SPAG6, a component of the central apparatus of the “9+2” axoneme, plays a central role in flagellar stability and motility. Immune cells lack cilia, but the immunological synapse is a surrogate cilium as it utilizes the same machinery as ciliogenesis including the nucleation of microtubules at the centrosome. We demonstrate that Spag6 localizes in the centrosome and is critical for centrosome polarization at and actin clearance away from the synapse between CTL and target cells. Furthermore, improper synapse formation and function likely explains reduced CTL function and class-switched antibody production in Spag6KO mice.

by Li Wen. / "April 2000." / Thesis (Ph.D.)--Chinese University of Hong Kong, 2000. / Includes bibliographical references (p. 200-236). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Mode of access: World Wide Web. / Abstracts in English and Chinese.

Measuring the Effects of Mouse Allergen and Black Carbon Exposure on Children Living in New York City with Allergic Diseases Medina Samira Jackson-Browne Background: Exposure to allergens and combustion by-products are risk factors for allergic health outcomes in children. The connection between exposure to allergens and allergic diseases such as asthma, in some children, is through the development of a biological condition known as allergic sensitization. In susceptible children, sensitization may occur when early-life exposure to an allergen causes the production of immunoglobulin E (IgE) antibodies. In asthmatic children, repeated exposures to this allergen may lead to clinical manifestations including airway inflammation, airway mucous production, bronchospasms, and bronchial hyper-responsiveness. Sensitization and repeated exposure to allergens may, therefore, be important risk factors for asthma morbidity in children. Findings from a cross-sectional asthma study of children living in NYC published previously by our group found a positive association between cockroach and dust-mite allergens measured in bed dust and sensitization risk to these allergens consistent with other studies. However, contrary to previously published research, no association was observed between mouse allergen measured in bed dust and mouse sensitization risk in our study. In urban areas such as New York City (NYC), exposure to combustion by-products, including black carbon (BC), has been shown to be associated with both asthma development and asthma morbidity. BC has been proposed to exacerbate asthma symptoms directly through airway irritation or by behaving as an adjuvant, enhancing the production of IgE antibodies following exposure to an allergen in sensitized individuals. Our group previously observed an association between indoor measured BC concentrations and airway inflammation, however no association was found between BC and asthma symptoms for children living in NYC. In the present study, we sought to address some of the limitations of the previous work. These limitations included a singular measurement of mouse allergen exposure, a shorter-term BC exposure measurement, and a cross-sectional study design for asthma symptom risks. My overarching hypothesis for this dissertation is that exposures to mouse allergen and BC are significant risk factors for allergic sensitization and asthma morbidity, respectively, for children living in NYC. I tested these hypotheses in three separate manuscripts by assessing multiple mouse exposure measurements with the risk for mouse sensitization (Chapter 2), testing the correlation between 7-day measured indoor BC and particulate matter smaller than 2.5 microns (PM2.5) concentrations with annual modeled outdoor BC and PM2.5 concentrations (Chapter 3), and determining whether annual modeled outdoor BC concentration is associated with persistent asthma symptoms, over a three-year period, for asthmatic children in NYC (Chapter 4). Methods: For all manuscripts, data from an asthma case-control cohort of children (age 7-8 years) previously established by our group, the NYC Neighborhood Asthma and Allergy Study (NAAS), was utilized for analysis (n=350). Kitchen floor and bed settled dust samples were collected from the children’s home during the initial home visit. Mouse allergen concentrations were quantified from both kitchen floor and bed dust samples using an enzyme-linked immunosorbent assay (ELISA). Blood samples were also collected during this visit. IgE antibodies to mouse allergens were measured by ImmunoCAP (Phadia, Uppsala, Sweden) from these blood samples. Information on the frequency of mouse sightings in the previous 12 months was extracted from a questionnaire administered to parents of NAAS children. Neighborhood and school mouse sightings were collected from reports from the NYC Department of Health and Mental Hygiene (DOHMH). Indoor PM2.5 and BC samples were collected from air samplers placed in NAAS children’s home for an average of 7 days. In collaboration with the NYC DOHMH, we were given access to 2-year averaged modeled outdoor PM2.5 and BC concentrations collected from air monitors at 124 street-level locations throughout NYC from 2008-2010. After the initial home visit, asthmatic NAAS children were followed-up annually for asthma symptoms. The questionnaire data collected from the asthmatics followed were used to evaluate the persistence or remittance of asthma symptoms over the 3-years following the initial home visit. Results: In our mouse study we found that increasing mouse allergen measured from kitchen floor dust and children whose parents reported greater than weekly mouse sightings in the previous 12 months has an increased risk of mouse sensitization (prevalence risk (PR) = 1.09 [1.02-1.17], p=0.04 and PR= 3.84 [1.95-6.97], p=0.001 respectively). Neither mouse allergen measured from settled bed dust (PR = 1.06 [0.95-1.19], p=0.46) nor neighborhood rodent reports (PR = 1.25 [0.94-1.68], p=0.16) were significantly associated with an increased risk of sensitization to mouse. Exposure to mouse at school was also not associated with an increased risk of mouse sensitization (PR=0.66 [0.35-1.26], p=0.30). Results from the correlation study indicated both annual modeled outdoor PM2.5 and BC concentrations were weakly correlated with 7-day measured indoor PM2.5 and BC concentrations (r = 0.21 and 0.39, respectively, p < 0.01). However, annual modeled outdoor BC concentrations predicted almost 20% of the variability of 7-day measured indoor BC (R2=0.19, p<0.001) compared to only 4% of the variability of 7-day indoor PM2.5 explained by annual modeled outdoor PM2.5, which predicted measured indoor PM2.5 (R2 = 0.04, p < 0.001). Our regression analysis of the asthma morbidity study found no significant association between longer-term neighborhood modeled BC concentrations at study participant’s home (PR = 0.87 [0.58-1.29, p=0.49] and school addresses (PR =1.09 [0.77-1.56], p=0.60) and persistent asthma symptoms. Conclusions: My findings suggest that mouse allergen measured from kitchen floor dust and parent reported mouse sightings are important risk factors of mouse sensitization for children living in urban areas such as NYC. The results of the BC analysis indicate a moderate correlation between annual modeled outdoor BC concentrations and 7-day measured indoor BC concentrations. The annual modeled outdoor BC also predicted 20% of the variability in 7-day measured indoor BC. Conversely, PM2.5 analysis indicate that annual modeled outdoor PM2.5 is not correlated with 7-day measured indoor PM2.5 concentrations. Finally, regression analysis of BC exposure and asthma morbidity indicate that annual modeled outdoor BC is not predictive of persistent asthma symptoms in our cohort.

Peanut allergy is a public health concern which affects a significant proportion of the population. Accidental exposure to peanut can cause severe and fatal reactions in peanut allergic individuals and currently their only safeguard is to practise careful avoidance. Identification and protection of at-risk members of the allergic population is critical in managing this life-threatening condition. This thesis produces key data to enable this. A prospective study was performed on 60 peanut allergic participants to determine thresholds of reactivity to peanut using oral challenges with incrementally increasing amounts of peanut protein. Following a double-blind placebo-controlled peanut challenge, participants received three further peanut challenges, two with co-factors: sleep deprivation and exercise, and one without. Severity was measured using a numerical scale derived from symptoms and serum tryptase was measured at each challenge. A total of 187 challenges were performed. Findings were that the median amount of peanut protein which induces a reaction in 10% of the population (ED10) was 12.3mg (95% CI 7.3,20.4) equivalently this suggests that 90% of the allergic population will not react to doses below this level. Both sleep deprivation and exercise have a significant effect on lowering reaction threshold (ED10), by 5 times and 2.5 times respectively. Separately there is a reduction in threshold with successive challenges. Co-factors also significantly increased symptom severity during challenge reactions. In particular sleep deprivation significantly increased the severity of gastrointestinal symptoms suggesting that a stressful stimulus may affect intestinal permeability. Evidence was provided for the importance of asthma as a risk factor which increased the severity of respiratory symptoms during reaction. Using a novel visual analogue scale for measuring the participant’s perception of severity, a poor correlation was observed between the participant’s perception of the reaction and the overall numerical severity score, suggesting that participants misperceive severe symptoms. This thesis provides the first data showing that symptom patterns in repeated challenges show a high degree of homogeneity within individuals, but importantly that this symptom homogeneity is also observed across individuals. Lastly the utility of serum tryptase in identifying food allergic reactions has been disputed previously. This thesis provides evidence of its value and identifies a rise cut-off of 30% as being diagnostic of a food allergic reaction, but cautions that acute levels must be compared with baseline as this rise may occur within the normal range.

Nas últimas décadas tem sido enfatizada a relevância do estudo das interações específicas entre os sistemas nervoso e imune. Pesquisas conduzidas ao longo dos últimos anos, desde Selye, têm demonstrado não apenas que o estresse modifica a atividade de órgãos linfóides, mas também que o sistema nervoso tem sua função modificada pelo sistema imune. A constatação das relevantes interações bidirecionais existentes entre os sistemas imune e nervoso, e da implicação destas interações na fisiologia e fisiopatologia acabaram por constituir o pilar mestre de um ramo da ciência conhecido como Neuroimunomodulação. Evidências recentes têm demonstrado um importante papel das vias colinérgicas parassimpáticas na comunicação bidirecional entre o cérebro e o sistema imune: a denominada Via Colinérgica Antiinflamatória. Há evidências na literatura de que as vias aferentes/eferentes do nervo vago são importantes nas interessantes interações que se estabelecem entre o sistema imune e áreas especificas do cérebro, tendo-se até mesmo sugerido que a estimulação elétrica de terminações eferentes do nervo vago teria ação terapêutica em processos inflamatórios, como, por exemplo, em modelos experimentais de sepsis, artrites, colites e pancreatites. Trabalhos experimentais de Basso et al. (2003), em nossos laboratórios, mostraram que uma reação alérgica intestinal altera a atividade de células do núcleo paraventricular do hipotálamo (PVN) e do núcleo central da amígdala (CeA), além de induzir alterações comportamentais indicativas de ansiedade. Neste contexto, pareceu-nos relevante avaliar uma possível participação do sistema nervoso autônomo parassimpático nas funções neurais e imunes desencadeadas por um processo alérgico intestinal. Mais especificamente verificar os possíveis efeitos da administração de anabasina na resposta imune entérica produzida pela administração oral de ovalbimuna (OVA) em animais OVA sensibilizados. / Studies of the specific interactions between nervous and immune systems increased in the last decades. Since Selye, it is known that stress change linphoids organs activity; and more recently it was shown that the nervous system activity might be changed by imune system products. The finding of the relevant bidirectional interactions between nervous and imune systems and of their implication on physiology and pathophysiology are now taken as the pillar of neuroimmunomodulation. Recent evidences are showning an important role for parasympathetic cholinergic pathways in the bidirectional communication between the brain and the immune system: the anti-inflammatory cholinergic pathway. Literature data showed that afferent/efferent pathways of the vagus nerve are relevant for the interactions between immune system and specific brain areas; these electric stimulation of the efferents fibers of vagus were reported to have a therapeutic effect on some inflammatory processes, such as colitis, arthritis and sepsis. Data from Basso et al. (2003) showed that an allergic intestinal inflammation changes cell activity within the paraventricular nucleus of the hypothalamus (PVN) and the central nucleus of amygdala (CeA), and also that it induces behavioral changes. Within this context, it seemed to us relevant to evaluate a possible Parassympathetic Nervous System participation on neural and immune functions alterations triggered by an allergic intestinal inflammation; specifically we thought it be interesting to study the effects of Anabasina administration, on enteric immune response triggered by oral administration of ovalbumine (OVA) in OVA-sensitized mice.

The aim of this thesis is to evaluate and optimise current pharmacotherapeutic options in rhinosinusitis. There is often a marked variation in treatment response in those afflicted with chronic rhinosinusitis, both within and between patients, attributable in part to different disease phenotypes/endotypes, poor awareness of treatment optimization options, and trivialization of symptoms by patients and physicians. Characteristically, these factors contribute to a typical remitting and relapsing disease course. The objectives of this work are to improve the therapeutic index and reach of commonly used medications by boosting efficacy whilst reducing concomitant side effects. The third chapter explores the use of initial oral steroids in patients with chronic rhinosinusitis and nasal polyposis, focusing on the role of the ostiomeatal complex in the perpetuation of disease symptoms. Often a short course of oral steroids is used in patients with moderate to severe disease to achieve initial control before maintenance with intranasal steroids. This is termed as a ‘medical polypectomy’ and anecdotally is commonly used in patients with chronic rhinosinusitis with nasal polyposis (CRSwNP). However, the evidence for its efficacy is tenuous and there are no data to evaluate if it indeed re-establishes ostiomeatal sinus complex drainage which is a condicio sine qua non of ensuring long-term symptom resolution. Further, it is known that monotherapy with nasal steroids may result in loss of symptom control. We have therefore in a double-blind placebo controlled trial (Chapter 4) evaluated the effect of this initial induction with oral steroids on subsequent sequential intranasal therapy. Perhaps, however, more crucially we have for the first time comprehensively addressed the safety of both oral and topical steroids in patients with CRSwNP who have other concomitant steroid-dependent illnesses like asthma and COPD. A particularly refractory subset of those with CRSwNP also have aspirin intolerance and asthma. While recent guidelines have recommended more aspirin challenge testing in these patients, it is unclear what the significance of a positive test is in the absence of overt clinical symptoms or in patients with only moderate disease. This is addressed in Chapter 5, as this significant phenotype of aspirin intolerant rhinosinusitis need close monitoring, dose optimization, polytherapy, and in selected cases may be suitable for aspirin desensitization. Penultimately, we evaluate in a double-blind placebo controlled trial (Chapter 6) the tachyphylaxis and rebound congestion that blights the medium to long-term use of sympathomimetic nasal decongestant sprays like oxymetazoline and if this can be reversed by the concomitant use of nasal steroids. We also characterized nasal blood flow as an outcome to evaluate in these patients and its relation to other rhinological outcome measures (Chapter 7).

Nas últimas décadas tem sido enfatizada a relevância do estudo das interações específicas entre os sistemas nervoso e imune. Pesquisas conduzidas ao longo dos últimos anos, desde Selye, têm demonstrado não apenas que o estresse modifica a atividade de órgãos linfóides, mas também que o sistema nervoso tem sua função modificada pelo sistema imune. A constatação das relevantes interações bidirecionais existentes entre os sistemas imune e nervoso, e da implicação destas interações na fisiologia e fisiopatologia acabaram por constituir o pilar mestre de um ramo da ciência conhecido como Neuroimunomodulação. Evidências recentes têm demonstrado um importante papel das vias colinérgicas parassimpáticas na comunicação bidirecional entre o cérebro e o sistema imune: a denominada Via Colinérgica Antiinflamatória. Há evidências na literatura de que as vias aferentes/eferentes do nervo vago são importantes nas interessantes interações que se estabelecem entre o sistema imune e áreas especificas do cérebro, tendo-se até mesmo sugerido que a estimulação elétrica de terminações eferentes do nervo vago teria ação terapêutica em processos inflamatórios, como, por exemplo, em modelos experimentais de sepsis, artrites, colites e pancreatites. Trabalhos experimentais de Basso et al. (2003), em nossos laboratórios, mostraram que uma reação alérgica intestinal altera a atividade de células do núcleo paraventricular do hipotálamo (PVN) e do núcleo central da amígdala (CeA), além de induzir alterações comportamentais indicativas de ansiedade. Neste contexto, pareceu-nos relevante avaliar uma possível participação do sistema nervoso autônomo parassimpático nas funções neurais e imunes desencadeadas por um processo alérgico intestinal. Mais especificamente verificar os possíveis efeitos da administração de anabasina na resposta imune entérica produzida pela administração oral de ovalbimuna (OVA) em animais OVA sensibilizados. / Studies of the specific interactions between nervous and immune systems increased in the last decades. Since Selye, it is known that stress change linphoids organs activity; and more recently it was shown that the nervous system activity might be changed by imune system products. The finding of the relevant bidirectional interactions between nervous and imune systems and of their implication on physiology and pathophysiology are now taken as the pillar of neuroimmunomodulation. Recent evidences are showning an important role for parasympathetic cholinergic pathways in the bidirectional communication between the brain and the immune system: the anti-inflammatory cholinergic pathway. Literature data showed that afferent/efferent pathways of the vagus nerve are relevant for the interactions between immune system and specific brain areas; these electric stimulation of the efferents fibers of vagus were reported to have a therapeutic effect on some inflammatory processes, such as colitis, arthritis and sepsis. Data from Basso et al. (2003) showed that an allergic intestinal inflammation changes cell activity within the paraventricular nucleus of the hypothalamus (PVN) and the central nucleus of amygdala (CeA), and also that it induces behavioral changes. Within this context, it seemed to us relevant to evaluate a possible Parassympathetic Nervous System participation on neural and immune functions alterations triggered by an allergic intestinal inflammation; specifically we thought it be interesting to study the effects of Anabasina administration, on enteric immune response triggered by oral administration of ovalbumine (OVA) in OVA-sensitized mice.

The first part of this research was undertaken to assess the impact of documented penicillin allergy on the choice of antibiotics and the clinical and financial consequences of changes in prescribing patterns in an Australian teaching hospital. The medical records of all patients aged >/= 18 years admitted with community-acquired pneumonia (CAP) to Sir Charles Gairdner Hospital (SGGH) over a 15-week period were reviewed prospectively. The severity of patients' penicillin allergies was assessed using a structured questionnaire. The antibiotic cost was calculated using acquisition, delivery (labour and equipment) and laboratory monitoring costs. The appropriateness of antibiotic prescribing was assessed using the Therapeutic Guidelines: Antibiotic (TG:A). The antimicrobial selections and costs were then compared for those patients with (Group A) and without (Group B) penicillin allergy. 155 patients were reviewed (males 71, females 84) with an average age of 68 ± 18 years. Of these, 27 (17.4%) had documented penicillin allergies; of which 12 were classified as Severity I (e.g. anaphylaxis, urticaria), 12 as Severity II (e.g. rash, itch) and three as intolerance (e.g. GI upset). The current TG:A recommends cephalothin or cephazolin as the drugs of choice for mild to moderate CAP patients with a history of penicillin allergy. However, combinations of cephalothin intravenously and azithromycin orally were the most commonly prescribed antimicrobials for such patients. The TG:A recommends erythromycin plus cefotaxime or ceftriaxone as the first-line therapy for severe CAP patients with a documented penicillin allergy. Yet, combinations of intravenous cephalothin, erythromycin and gentamicin were the most frequently prescribed antimicrobials for such patients. / A history of penicillin allergy significantly (p<0.05) increased the cost of antibiotic treatment and total cost of admission. The adherence of antibiotic prescribing to the TG:A for patients with penicillin allergies is variable. Patients with labelled penicillin allergies had greater antibiotic costs and total cost of admission. Identifying patients with intolerance rather than allergies would reduce the total inpatient costs at SCGH by A$ 463.01 a year for mild to moderate CAP patients and A$ 39 614.54 a year for severe CAP patients. The second part of the project was a prospective study of patients admitted to SCGH who had a history of penicillin allergy, but were not suffering from CAP. This study was conducted in order to ensure that the pattern of penicillin allergies of patients admitted to the hospital could be adequately characterised. Over a 5-week period, all adult patients admitted without CAP to SCGH who claimed to have a history of penicillin allergy were interviewed with regard to their penicillin allergies. The standard of allergy documentation was also assessed for each patient. Of the 140 patients assessed (males 63, females 77, average age 61 ± 17 years), 108 (77.1%) were classified as allergic: 61 (56.5%) as Severity I and 47 (43.5%) as Severity 11, 26 (18.6%) as intolerant and the remaining six (4.3%) as not substantiated. / The standard documentation of the patients' penicillin allergies was poor - only 40 (38.6%) of either medical records or drug charts had the type of reaction and only five (3.6%) had the date of reaction. In general, penicillin allergies were poorly documented in both patients' medical records and on drug charts. Inadequate detail of reported reactions often made it difficult to assess their clinical significance. These findings prompted a recommendation that pharmacists should help to ensure accurate allergy documentation by evaluating patients and educating both patients and health care professionals.