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DNA Sequence and Haplotype Variation Analysis of Inflammatory Response Genes NLRX1, IL6, and IL8 in the Turkey (Meleagris gallopavo)Russell, Kadijah Lashunte 08 February 2019 (has links)
Genotype-phenotype analyses continue to be the primary goal for genome analyses in livestock and poultry breeding. Essential to accomplish this goal is the need to identify variation at the genomic level. To test the hypothesis that DNA sequence variations in inflammatory response genes are associated with phenotypic differences in the heritage turkey, the primary objective of this project was to search for single nucleotide polymorphisms (SNPs) in candidate inflammatory response genes. A minor objective was to develop a system for inducing inflammatory response in the turkey using a microbe-based lipopolysaccharide (LPS), an approach previously described for the chicken. A total of 18 SNPs was identified in the three genes screened in this project: Interleukin 6 (IL6) and 8 (IL8), and NLRX1. Mortality data from the LPS challenge were not significantly different among the strains. Further gene expression analyses will be part of future work. The SNP data represent the first extensive analyses of candidate inflammatory response genes in the turkey. Combined with the protocols developed for inflammation assessment in the turkey the SNPs described here will be valuable resources for future inflammation:genotype evaluation in the turkey / MS / Though progress has been made in the genome analyses of the turkey, Meleagris gallopavo, our understanding of the genotype: phenotype relationships continue to lag those of agriculturally important animal species. Among the phenotypes for which genetic understanding can be useful is inflammation, a complex trait that is influence by many interdependent response mechanisms. These mechanisms, regarding differences across heritage turkeys, has been mildly investigated. Since Nucleotide Polymorphism (SNP) screening is a common method used to comprehend the robust effects these differences have on genotype and phenotype. Here, we report initial investigations in our lab of the genetics of inflammation in the turkey using comparative information from the chicken NOD like receptor X1 (NLRX1), turkey interleukin 6 (IL6), and Interleukin 8 (IL8). These genes were screened for nucleotide variants that may be informative for future studies that will investigate the turkey’s response to Salmonella derived lipopolysaccharide that can induce inflammation. The rationale for selecting these three genes is that IL8, IL6, and NLRX1 have pro inflammatory and/or anti-inflammatory functions that respond to maintain homeostasis. Primers were designed and investigated using DNA from Broad Breasted White (BBW), and Broad Breasted Bronze (BBB), Blue slate (SL), Spanish Black (SBL), Midget White (MW) and Royal Palm (RP). The birds were also challenged with 1.5 mg/kg Lipopolysaccharide (LPS) intra-abdominally to collect tissue post LPS challenge. Tissues was collected from the thymus, spleen, and bursa of fabricius: organs identified to effect inflammation. A total of 2,239 bp for IL8, 2,439 bp for IL 6, and 572 bp for NLRX1 were screened for SNPs. SNP analysis revealed 16 SNPs in the inflammatory response genes mentioned.
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The role of interleukin-12 in the pathogenesis of human systemic lupuserythematosus劉鐵夫, Liu, Tiefu. January 1998 (has links)
published_or_final_version / Pathology / Doctoral / Doctor of Philosophy
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The role of IL-6 in the immune response to coccidiaLynagh, Gail Rosemary January 1998 (has links)
No description available.
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IL7 receptor signalling during B cell developmentSmart, Fiona May January 1998 (has links)
No description available.
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Etude des mécanismes immunitaires dans un modèle d'inflammation pulmonaire allergique chez la souris : rôles de l'interleukine-22 / Roles of interleukin-22 in a mouse model of allergic airways inflammationBesnard, Anne-Gaëlle 17 December 2010 (has links)
L’asthme est une maladie inflammatoire chronique des voies aériennes. Chez les individus sensibles, l’inhalation d’allergènes entraine une inflammation pulmonaire se traduisant par des épisodes récurrents de toux, de difficultés respiratoires et une sécrétion de mucus. Des études réalisées chez l’animal ont mis en évidence un rôle crucial des lymphocytes Th2 et des cytokines associées (IL-4, IL-5 et IL-13). Plus récemment, il a été montré que les lymphocytes Th17 participaient à la physiopathologie de l’asthme. La présente étude s’intéresse à une cytokine majoritairement produite par les Th17 : l’IL-22. Différents travaux indiquent que cette cytokine serait impliquée dans l’immunitémucosale où elle exercerait des effets protecteurs ou pro-inflammatoires en fonction du modèle expérimental étudié. En utilisant un modèle murin d’inflammation pulmonaire allergique induite par l’ovalbumine, nous avons montré que l’IL-22 jouait un rôle pro-inflammatoire au cours de l’induction de l’asthme allergique puisque les souris déficientes en IL-22 développent une forme atténuée de la maladie. A l’inverse, nous avons constaté que l’IL-22 avait un effet protecteur dans la phase effectrice, et que cet effet était dépendant de l’IL-17A. Nos travaux mettent donc en lumière une double fonction de l’IL-22 dans l’asthme allergique chez la souris. En parallèle de ce travail, nous nous sommes intéressés au rôle de l’IL-1 et de l’inflammasome NLRP3 dans ce même modèle d’inflammation pulmonaire. Enfin, une troisième étude a permis de mettre en lumière un rôle encore inconnu de l’interleukine-33 dans l’activation des cellules dendritiques au cours de la mise en place de la réponse asthmatique. / Asthma is a heterogenous inflammatory disorder of the airways characterized by chronic airway inflammation, airway hyper-reactivity and by symptoms of recurrent wheezing, coughing and shortness breath. Understanding of the role of allergy and Th2 cells in asthma has benefited from mouse model of allergic asthma. Recently, several studies highlighted Th17 involvement in asthma pathogenesis. In the present study, we investigate the role of IL-22, a Th17-related cytokine, in a mouse model of allergic lung inflammation induced by ovalbumin. First, using IL-22 deficient mice, we demonstrated a pro-inflammatory role of IL-22 during the sensitization phase. In contrast, we observed a protective function of IL-22 during the effective phase. This protective effect of IL-22 seems to be dependent of IL-17. In conclusion, we demonstrate here a dual role of IL-22 in asthma pathogenesis. Since interleukin-1_ is critical for Th17 polarization in human, we also investigated the role of IL-1 signalling and NLRP3 inflammasome in our model of allergic airway inflammation. We showed that NLRP3 inflammasome and IL-1R/IL-1 pathway are critical to induce allergic lung inflammation, even in the absence of adjuvant. Finally, we studied the effect of interleukin-33 on dendritic cells activation and Th2 priming during antigen sensitization and in established asthma.
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The role of interleukin-8 in the immunopathogenesis of HIV-1 disease and tuberculosisMeddows-Taylor, Stephen 27 May 2014 (has links)
Interleukin-8 (IL-8), a member of the C-X-C chemokine subfamily, is an important
chemoattractant and cellular activator. This study was conducted to determine the role of IL-8
in the immunopathogenesis of HIV-I disease and tuberculosis.
The first section involved determining the effect of infection with HIV-1, Mycobacterium
tuberculosis and co-infection with both of these organisms on IL-8 j_ roduction in vivo. This was
monitored by the determination of levels of serum or plasma EL-8 and peripheral cell-associated
IL-8, assessing peripheral mononuclear (PBMC) and polymorphonuclear (PMN) cell capacity to
produce IL-8 spontaneously or in response to various stimuli, and the detection of constitutive
IL-8 mKNA expression in purified subsets of mononuclear cells. Results show that whereas there
is evidence of detectable levels of cell-associated EL-8 (mKNA and protein) in peripheral cells of
healthy individuals, this is largely lost in the disease states studied. Coupled with this was
significantly increased circulating levels of EL-8 in serum and plasma found in HIV-1 infected
individuals with or without concomitant pulmonary TB. On the other hand, the capacity of PBMC
to produce IL-8 spontaneously ex vivo was enhanced in HIV-1 and TB patients and many of the
HFV/TB group, but their corresponding capacities to respond to various stimuli was significantly
diminished when compared to that of the normal donors. The release of IL-8 from PMN in the
presence of an agonist was diminished mainly in individuals with pulmonary TB, which was
further exacerbated by the presence of HIV-1 infection.
HIV-1-infected individuals have an increased incidence of bacterial infections which could
be related to defective functioning of PMN. The second section was aimed at detecting PMN
abnormalities in HIV and I-HV/TB patients by monitoring EL-8-induced p-glucuronidase release
and PMN chemotaxis in response to IL-8. IL-8-induced (I-glucuronidase release from PMN of
normal individuals and TB patients occurred in a dose-dependent manner. In contrast, PMN from
HTV-1 infected individuals, whether co-infected with M tuberculosis or not, showed a reciprocal
response in that increasing IL-8 concentrations resulted in decreased enzyme release. This
reciprocal slope of the IL-8 dose-response curve was altered for the majority of HIV-1 positive
individuals tested irrespective of their CD4+ cell counts. In addition, PMN chemotaxis in response
to IL-8 was also found to be significantly impaired in a group of HIV-1 infected patients coinfected
w ithM tuberculosis when compared to healthy individuals.
The third section of the study involved analysing the expression of the PMN cell surface
markers, FcyRIII (CD 16), and the two human IL-8 receptors, designated IL -8RA and 1L-8RB.
FcyRIII (CD 16) expression on the surface of PMN was significantly reduced in HIV-1
seropositive patients with pulmonary tuberculosis when compared to those individuals with either
disease alone or healthy blood donors. A significant reduction in the percentage of PMN
expressing IL-8RA and IL-8RB and in their respective fluorescence intensities was found in TB,
HIV, and HTV/TB groups when compared to that obtained for the ND group. IL-8RA intensity
of fluorescence was significantly decreased in the HTV/TB group when compared to the TB and
HIV groups indicating a further down-regulation of IL-8RA expression owing to dual infection.
On the other hand, IL-8RB fluorescence intensity was substantially reduced on PMN from
patients with pulmonary TB and to a greater degree in those patients co-infected with HIV-1 and
M. tuberculosis. Having found a reduction in the expression of both IL-8 receptors on PMN in
all the infection groups, cellular events following the binding of IL-8 to IL-8 receptors on PMN
isolated from dually infected patients, the group which showed the greatest reduction in IL-8
expression was analysed. Results indicated that the impairment of DL-8-dependent PMN functions
such as degranulation and chemotaxis was associated with the reduced expression of IL-8
receptors on these cells.
Increased circulating levels of IL-8 in HIV-1 infection and a diminished cellular capacity
to produce IL-8 as shown in this study may have important implications for antimicrobial defences
and normal immune processes. A dysregulated production of IL-8 in vivo is likely to play a role
in the pathogenesis of HIV-1 disease, pulmonary tuberculosis, and dual infections with both
organisms. In addition, cellular responses dependent on specific receptor engagement and the
subsequent translation of signal transducing events that lead to phagocyte effector functions are
clearly impaired in IL-8 receptor deficient phagocytes. Abnormal PMN functioning in HTV-1
infected individuals, as shown here by defective degranulation and chemotactic responses, have
important implications in the pathogenesis of HIV-1 infection in terms of their ability to clear
secondary microbial infections. Future attempts should be aimed at defining the mechanisms that
bring about these changes in order to contribute to a greater understanding of the mechanisms that
lead to an enhanced risk of superinfections in immunosuppressed individuals.
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Alternative activation of dendritic cellsJones, Lucy Helen January 2013 (has links)
The alternative activation of macrophage populations by Interleukin-4 (IL-4) is well characterised. Alternatively activated macrophages (AAM) express high levels of the arginine converting enzyme arginase-1, and express a plethora of IL-4 driven molecules including the resistin like molecule alpha (RELMα) and the chitinase like molecule Ym1/2. Dendritic cells (DCs) are the professional antigen presenting cells (APC) of the immune system, responsible for the detection of invading pathogens, secretion of cytokines and the subsequent activation of T-cells. This thesis addresses whether IL-4 is able to ‘alternatively activate’ DCs both in vitro and in vivo, in a manner similar to that of AAM. The impact of IL-4 on DC and macrophage activation was compared and contrasted, and it was confirmed for the first time that IL-4 can alternatively activate DCs, inducing high level expression of a range of alternative activation associated markers including RELMα, Ym1/2, CCL24 and dectin-1, with the exception of arginase. DCs were significantly more capable at the in vivo priming of T-cell responses in the context of both Th1 and Th2 polarising antigens than similarly exposed macrophages, confirming their superior capacity as APC. The requirements for DC IL-4Rα expression were assessed, and IL-4 responsiveness was found to be required for the optimal induction of Th1 responses. Conversely, selective loss of only one facet of the IL-4 response, namely RELMα expression, limited the ability of IL-4 exposed DCs to induce the regulatory cytokine IL-10 both in vitro and in vivo. Furthermore, alternatively activated DCs (AADCs) were found in the spleen following 8 weeks of infection with the parasitic trematode Schistosoma mansoni, highlighting a role for DC alternative activation in a disease setting. IL-4 was shown to induce expression of the vitamin A converting enzyme aldehyde dehydrogenase, and the product of such activity, retinoic acid (RA), was found to promote the expression of RELMα in IL-4 exposed DCs. Aldehyde dehydrogenase activity was found to inversely correlate with DC expression of Ym1/2 and inhibition of RA signalling limited IL-4 driven RELMα and promoted Ym1/2.
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The Blimp-1-Dependent Interleukin-2 Inhibitory Loop in CD4+ T cellsOuyang, Li 01 January 2008 (has links)
IL-2 has multiple functions in T cell-mediated adaptive immunity. The stringent control of its expression is important for T cell activation, proliferation and the subsequent T cell clone contraction. Our lab has recently shown that the transcriptional repressor Blimp-1 is part of a negative feedback loop which controls IL-2 gene expression in mice. Understanding the molecular mechanisms of this signaling loop in T cells might help us to better understand the regulation as well as the role of IL-2 in T cell immunity. The human ortholog to murine Blimp-1 is termed PRDI-BF1 (each encoded by the respective Prdm1 gene). Both genes contain five zinc finger regions, whereby the first two zinc fingers are dispensable for DNA binding. In case of the human protein they are instead required to recruit the G9?Ñ methyltransferase to the gene promotor. We found that the human wild-type PRDI-BF1 protein suppressed IL-2 production in murine T cells, while deletion of the first two zinc fingers abolished this ability. Thus, a similar Blimp-1-mediated methylation mechanism might exist in IL-2 gene silencing. IL-2/IL-2R signaling is indispensable for Blimp-1 induction. PI-3Kinase and Stat5 are downstream of the IL-2 receptor complex and are known to contribute to IL-2 inhibition in T cells from C57BL/6 mice. However, activating only these two pathways are still not sufficient to induce Blimp-1 or suppress IL-2 expression in in IL-2R beta-/- mice. The Blimp-1-dependent IL-2 self regulatory loop is not functional in IL-2R beta-/-mice. In order to conveniently study this dysregulation we crossed these mice with a GFP transgenic strain in which the GFP transgene is under the control of IL-2 promoter sequence. In IL-2R beta-/-IL-2p-GFP mice about five times as many spleenic CD4+ T cells transcribe IL-2pGFP, compared to the littermate IL-2R beta+/-IL-2p-GFP control animals. And most of the GFP cells demonstrate activated phenotype (CD44HighCD62Llow). Blimp-1 is known as a master regulator of B cell terminal differentiation. Since a recent report indicated that IL-2 signaling via STAT5 constrains Th17 Cell differentiation, we speculated that Blimp-1 might play a similar role in effector T cell differentiation. In order to evaluate this possibility, activated CD4+ T cells from C57BL/6 mice were transduced with Blimp-1 and cultured under Th17 polarizing conditions. Blimp-1 overexpression in did not change the profile of IL-17 production.
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Inflammatory Mechanisms After Thromboembolic Ischemic Stroke in MiceAbulafia, Denise P. 12 June 2008 (has links)
Stroke induces multiple pathological sequelae directly affecting neuronal survival and eliciting short and long-term deficits in behavioral outcome. Most of stroke models utilized to investigate these pathological consequences are based on pure cerebral ischemia models. However, human thromboembolic stroke is characterized by a complex multifactorial response that involves the activation of the cerebral microcirculation by the occluding thrombus. Here, we have characterized a novel mouse model of tromboembolic stroke that mimics most of the clinical aspects of the human pathology. The common carotid artery thrombosis (CCAT) model produces consistent and reproducible infarcts and triggers an inflammatory response comparable to other well established models of stroke. Several of the pathological consequences of cerebral ischemia are triggered by focal inflammatory processes that occur early after the ischemic event. Cerebral inflammation is initiated by an early release of pro-inflammatory cytokines. These active cytokines promote the recruitment of inflammatory cells from the blood cerebral circulation into the brain parenchyma and subsequent release of additional amounts of inflammatory cytokines. This exacerbated cytokine response result in further irreversible neuronal and histopathological damage. Cytokines interleukyne-1 beta (IL-1 beta) and interleukyne-18 (IL-18) maturation requires the presence of active caspase-1. Activation of caspase-1 in the peripheral immune response involves the recruitment of several caspase-1 molecules into a macromolecular complex termed the inflammasome. Cerebral ischemia triggers the synthesis and activation of caspase-1. However, the cellular mechanisms associated to the activation of caspase-1 in the ischemic brain remain to be elucidated. In this study, we demonstrate that the NLRP1-inflammasome composed by capase-1, ASC (apoptosis-associated speck-like protein containing a caspase-activating recruitment domain) and NLRP1 (NLR (nucleotide binding, leucine-rich repeat) is assembled following the ischemic event. Moreover, we have characterized the cellular distribution of the inflammasome proteins in the normal and the ischemic brain. Data from this investigation suggest that six to twenty four hours following CCAT the inflammasome complex is assembled in neurons while microglia, macrophages and astrocytes form this complex at 7 days following cerebral ischemia. On the basis of these findings we next investigated whether inhibition of the inflammasome complex reduces the inflammatory response after ischemia. Neutralization of NLRP1 utilizing a specific antibody, revealed decreased activation of caspase-1 and IL-1 beta and reduced histopathological damage within the ischemic brain. Thus, the inflammasome complex is a major contributor of the inflammatory response following cerebral ischemia and inhibition of this complex may be a novel therapeutic target for reducing the pathological consequences of stroke.
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Development of an interleukin 2 receptor targeted gene therapy vehicleWattanakaroon, Wanida 16 August 2006 (has links)
The effectiveness of most chemotherapeutic regimens is limited by the toxicity of
the therapy to normal healthy cells. Therapies to selectively modulate abnormal T cells
bearing the interleukin 2 receptor (IL-2R) have been developed to treat diseases
associated with aberrant immune response. This study describes the development and
optimization of a targeted gene or oligonucleotide therapy vehicle to IL-2R bearing T
cells for selective elimination of these cells. In this work, a monoclonal antibody to the
IL-2R was used to target the oligonucleotide delivery vehicle which consisted of a
polyamidoamine dendrimer. Optimization of the delivery vehicle involves
understanding the factors that govern its association with oligonucleotide, the pathway
of IL-2R endocytic trafficking, and the stability of the oligonucleotide in the biological
milieu. Oligonucleotide stability in a cellular environment was examined intra- and
extracellularly. Results showed that the rate of intracellular degradation of
oligonucleotides was much greater than extracellular degradation. Binding of
oligonucleotides to dendrimers was demonstrated as a function of dendrimer generation.
The total binding capacities for dendrimers differed depending upon dendrimer size and
surface group, whereas equilibrium binding affinity was comparable for all dendrimers
tested. Binding of oligonucleotide delivery vehicle to the cell surface and subsequent
internalization was inversely related to dendrimer size, and in all cases, significantly less
than binding and internalization of the natural ligand for the IL-2R. Based on
experimental results, a kinetic model of the delivery vehicle was derived which includedthe dependence of binding and internalization on dendrimer size and surface charge and
intracellular degradation of oligonucleotide. Based on model predictions, we show that
larger dendrimers carry more oligonucleotide than the smaller dendrimer vehicles, and
delivery is more effective with larger vehicles. This work establishes our ability to
predict the effects of different delivery vehicle properties on oligonucleotide delivery
and aids in the development of design criteria for new vehicles for delivery of antisense,
siRNA, or genes to IL-2R bearing cells.
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