IL-1 is an essential pro-inflammatory factor in inflammation response. The effect of IL-1 is through binding to the IL-1 receptor that triggers the following signal transduction pathway. To study the role of IL-1 receptor-mediated signal pathway in inflammatory response, injecting P. aeruginosa into trachea of wild-type (WT) and type-1 IL-1 receptor knock-out (IL-1R1-/-) mice was used as the experimental model. Injecting bacterium into trachea of mice will induce pneumonia which increases accumulation of neutrophils, production of nitric oxide, expression of intercellular adhesion molecule-1 as well as many kinds of cytokines and causes the lung damage. The pneumonia-induced lung damage and inflammation at 24 hr after injecting P. aeruginosa into trachea were more severe in knock-out than in WT mice, as demonstrated by increases in extravasations of Evans blue dye (EBD), myeloperoxidase (MPO) activity, expression of iNOS, IL-1 beta and ICAM-1, and higher mortality of knock-out mice. The cause of the high mortality in knock-out mice was further investigated by culturing the lung and blood samples for bacterial counts. The bacterial counts of lung and blood of IL-1R1-/- mice were all higher than that of WT mice in 8 to 24 hr after injection of bacterium. Finally, chimeric mice (WT ¡÷ WT, IL1R1-/- ¡÷IL1R1-/-, WT ¡÷ IL1R1-/-, IL1R1-/- ¡÷ WT) were generated and used to determine the role of PMN cells of blood. Suggesting that increased amounts of bacteria in lung and blood is related to the higher mortality in knock-out mice and the type-1 IL-1 receptor is essential for mice to against pneumonia in this model.

Thesis (M.Sc.). / Written for the Dept. of Biochemistry. Title from title page of PDF (viewed 2008/05/29). Includes bibliographical references.

The oncogenic role of many of inflammatory genes in prostate cancer (PCa) remains unexplored despite the increasing association of chronic inflammation with PCa initiation, progression, and therapy resistance. The overarching goal of this project was to identify dysregulated inflammatory genes that correlate with PCa progression and seek to understand their molecular mechanisms and the therapeutic potential of targeting them. To achieve this, we utilized cutting-edge integrative (epi) genomic and transcriptomic techniques to identify and characterize inflammatory genes whose deregulation or (epi) genetic alterations correlate with PCa progression. Weighted Gene Co-expression Network Analysis and other multivariate analysis techniques identified IRAK1 as one of the inflammatory signatures found to be overexpressed in over 80% of prostate adenocarcinoma (PRAD) samples. We also explored the diagnostic and prognostic potential of IRAK1 as a biomarker using Kaplan Meier Survival Analysis and AUROC Analysis. DNA methylation analysis showed that IRAK1 is hypomethylated and found to negatively correlate with its overexpression in PRAD patients. We also found some missense and truncated mutations in some patients and reported a high level of IRAK1 gene amplification in castration-resistant and neuroendocrine PCa patients. / Includes bibliography. / Dissertation (PhD)--Florida Atlantic University, 2021. / FAU Electronic Theses and Dissertations Collection

No description available.

Le domaine TIR (Toll/interleukin (IL)-1 receptor) est une composante essentielle du système immunitaire inné, celui-ci est présent dans les récepteurs TLR (Toll-like receptor) et les protéines adaptatrices associées comme MyD88 et TIRAP. La détection de pathogènes déclenche l'interaction entre les domaines TIR permettant ainsi l'initiation et la propagation de la signalisation par les TLRs. Aussi, de nombreux pathogènes produisent des effecteurs contenant un domaine TIR tels que BtpA et BtpB chez Brucella abortus, TirS chez Staphylococcus aureus ou TcpC chez l'uropathogènique Escherichia coli. Tous ces effecteurs bloquent la signalisation des TLRs et sont capables de perturber les voies de signalisation de l'immunité innée pendant l'infection. Cependant les mécanismes moléculaires impliqués restent la plupart du temps non caractérisés et dans certains cas controversés. Dans le but de mieux comprendre le fonctionnement de ce type d'effecteurs bactériens, j'ai caractérisé chez Pseudomonas aeruginosa PA7 un nouvel effecteur contenant un domaine TIR que nous avons renommé PumA pour Pseudomonas UBAP1 Modulator A. En parallele, j'ai aussi participé à des projets de caractérisation de deux autres effecteurs avec un TIR domain : BtpB et TirS. Ainsi, PumA est un facteur essentiel pour la virulence de P. aeruginosa PA7 et son domaine TIR est essentiel pour interaction avec deux protéines adaptatrice, TIRAP et MyD88. Durant l'infection de cellules épithéliales pulmonaires par P. aeruginosa PA7, PumA est responsable du contrôle de la translocation du facteur de transcription NF-κB dans le noyau. De plus, la production de PumA dans une souche de P. aeruginosa non-TIR confère à cette bactérie de nouvelles propriétés d'immuno-modulation. PumA cible aussi UBAP1, une protéine du complexe de tri endosomal requis pour le transport, ESCRT-I (endosomal sorting complex require for transport I) qui a été récemment montré pour moduler l'activation de récepteur de cytokine. Nos résultats montrent que UBAP1 peut s'associer avec TIRAP et MyD88, provoquant le mouvement de MyD88 à la membrane cytoplasmique, suggérant une nouvelle voie cellulaire commune entre UBAP1 et les TLRs, et révélant UBAP1 comme nouvelle cible pour des effecteurs bactériens dans le cadre du contrôle des réponses immunitaires de l'hôte / In higher eukaryotes, the innate immune system provides the first line of defense against invading pathogens. The Toll/interleukin-1 receptor (TIR) domain is an essential component of the innate immune system. This domain is present in Toll-like receptors (TLRs) and associated adaptor proteins such as MyD88 and TIRAP. Pathogen detection requires interaction between the TIR domains, which initiates and triggers propagation of TLR signaling. However, many pathogens produce a TIR domain-containing protein such as BtpA and BtpB in Brucella abortus, TirS in Staphylococcus aureus or TcpC in the uropathogenic strain Escherichia coli. These effectors block TLR signaling and are able to disrupt innate immune response during infection. However, the molecular mechanisms involved remain mostly uncharacterized and in some cases controversial. The objective of this thesis was to study bacterial effectors containing a TIR domain particularly at the molecular level. For this, we focused on Pseudomonas aeruginosa PA7, an atypical multi-drug resistant strain that contains an effector with a TIR domain that we named PumA, for Pseudomonas UBAP1 Modulator A. In addition, during these four years of thesis work I also participated in the characterization of two other effectors with a TIR domain: BtpB in B. abortus and TirS in S. aureus.We found that PumA is essential for virulence of P. aeruginosa PA7 and its TIR domain is the key element for interaction with two adaptor proteins MyD88 and TIRAP. During infection of lung epithelial cells by P. aeruginosa PA7, PumA is responsible for controlling the translocation of NF-?B into the nucleus indicative of activation of this transcription factor. In addition, production of PumA by a TIR-deficient strain of P. aeruginosa confers to this bacterium a new immuno-modulation property. Furthermore, PumA targets ubiquitin-associated protein 1 (UBAP1), a protein of the endosomal sorting complex required for transport I (ESCRT-I) which has recently been shown to modulate cytokine receptor activation. Our results also show that UBAP1 can associated with TIRAP and MyD88, causing movement of MyD88 to the cytoplasmic membrane and suggesting a new cellular pathway between UBAP1 and TLRs. In summary, our data reveal UBAP1 as a novel target for bacterial effectors implicated in control of host immune responses

Immunological stressors, in the form of clinical and sub-clinical disease are currently controlled using both prophylactic antibiotics in-feed, and therapeutic antibiotic treatment. Respiratory disease, primarily Actinobacillus pleuropneumoniae (App) infection, is recognised as a major factor causing reduced productivity in pigs. This thesis reports investigations into the use of novel immunomodulators in particular Interleukin 4 (IL-4) and Interleukin 1 receptor antagonist (IL-1ra) as alternatives to antibiotics to treat App infection. Immunological and molecular biological assays were used to investigate and accumulate data. An in vitro study undertaken to find potential anti-inflammatory substances, revealed that Interleukin 8 (IL-8) mRNA production stimulated by PMA or LPS in whole pigs' blood was suppressed by IL-4. IL-1ra also suppressed stimulated IL-8 mRNA production by heat killed App bacteria (KB) in vitro. An acute LPS challenge in pigs in vivo however, showed no variation in illness or weight loss between pigs treated prophylactically with anti-inflammatory substance (IL-4 and IL-1ra) and saline treated pigs. The use of plasmids as a delivery system for anti-inflammatory substance did not show promise since it did not enhance growth or prolong the expression of the substances in the pigs. However, in the chronic App challenge model IL-4 and IL-1ra administered prophylactically in vivo showed an ability to improve growth. The therapeutic administration of IL-4 and IL-1ra to App challenged pigs showed no difference in pigs' growth, regardless of the treatment or control administered. To conclude, IL-4 and IL-1ra showed promise when administered prophylactically and improved growth and abrogated disease under conditions of App challenge. However when IL-4 and IL-1ra where administered therapeutically they did not perform as well. Moreover these compounds have potential as a commercial application to reduce the growth reduction caused by disease such as App.

Interleukin-1 (IL-1) is a prototypic multifunctional cytokine. IL-1 family include interleukin-1 a (IL-1 a), interleukin-1b (IL-1 b) and interleukin-1 receptor antagonist (IL-1 Ra). IL-1 b is the archetypeal pleiotropic cytokine which have been produced by many cells and exerting its biological effects on almost all cell types. IL-1 b is the most potent of known agents that are gastric cytoprotective, antiulcer, antisecretory and an inhibitor of gastric emptying. IL-1 Ra competes with IL-1 b for cell surface receptor occupancy. Host genetic factors that affect interleukin-1 (IL-1) have been reported to influence the susceptibility of Caucasians to gastric cancer. Whether Asians have the same genetic susceptibility remains unclear. In this study, the genetic associations of IL-1B and IL-1RN polymorphisms with gastric cancer and duodenal ulcer in Taiwan were evaluated. Genomic DNA from 140 unrelated Taiwanese patients with gastric adenocarcinoma, 94 with duodenal ulcer and 165 ethically matched healthy controls was typed for polymorphisms at positions ¡V31, -511, and +3954 in the IL-1B gene, and the variable number of tandem repeats polymorphisms in intron 2 of the IL-1RN gene. The allele frequencies of IL-1RN 2R in gastric cancer cases were much higher than those in healthy controls (9% vs. 3%, p = 0.781). The allele frequencies of IL-1B ¡V31, IL-1B ¡V511 and IL-1B +3954 did not differ. An increased risk of the development of intestinal type gastric carcinoma was found in IL-1RN 2R carriers with an odds ratio (OR) of 4.06 (95% confidence interval [CI]: 1.68 ¡V 9.79, p-value=0.085). And another increased risk of the development of diffuse type gastric carcinoma was found in IL-1RN 2R carriers with an odds ratio (OR) of 3.15 (95% confidence interval [CI]: 1.16 ¡V 8.56, p-value=0.061). A significant association was found in IL-1RN 2R/4R genotype and the risk of the development of duodenal ulcer, with an odds ratio (OR) of 2.57 (95% CI: 1.03 ¡V 6.38, p = 0.292). No significant relationship was noted in duodenal ulcer patients with IL-1B genotype examed in this study. Additionally, a synergistic interaction between blood type A and IL-1 RN 2R carriers existed in gastric cancer patients (OR= 4.51; 95% CI: 1.20 ¡V 16.88, p-value=0.516). The synergistic interaction was even stronger between blood type O and IL-1 RN 2R carriers of duodenal ulcer patients (OR= 10.3; 95% CI: 2.10 ¡V 50.61, p-value=0.160). In conclusion, the genetic polymorphisms of IL-1RN 2R and blood type A are associated with the development of gastric cancer. The genetic polymorphisms of IL-1RN 2R and blood type O are associated with the development of duodenal ulcer.

Immunological stressors, in the form of clinical and sub-clinical disease are currently controlled using both prophylactic antibiotics in-feed, and therapeutic antibiotic treatment. Respiratory disease, primarily Actinobacillus pleuropneumoniae (App) infection, is recognised as a major factor causing reduced productivity in pigs. This thesis reports investigations into the use of novel immunomodulators in particular Interleukin 4 (IL-4) and Interleukin 1 receptor antagonist (IL-1ra) as alternatives to antibiotics to treat App infection. Immunological and molecular biological assays were used to investigate and accumulate data. An in vitro study undertaken to find potential anti-inflammatory substances, revealed that Interleukin 8 (IL-8) mRNA production stimulated by PMA or LPS in whole pigs' blood was suppressed by IL-4. IL-1ra also suppressed stimulated IL-8 mRNA production by heat killed App bacteria (KB) in vitro. An acute LPS challenge in pigs in vivo however, showed no variation in illness or weight loss between pigs treated prophylactically with anti-inflammatory substance (IL-4 and IL-1ra) and saline treated pigs. The use of plasmids as a delivery system for anti-inflammatory substance did not show promise since it did not enhance growth or prolong the expression of the substances in the pigs. However, in the chronic App challenge model IL-4 and IL-1ra administered prophylactically in vivo showed an ability to improve growth. The therapeutic administration of IL-4 and IL-1ra to App challenged pigs showed no difference in pigs' growth, regardless of the treatment or control administered. To conclude, IL-4 and IL-1ra showed promise when administered prophylactically and improved growth and abrogated disease under conditions of App challenge. However when IL-4 and IL-1ra where administered therapeutically they did not perform as well. Moreover these compounds have potential as a commercial application to reduce the growth reduction caused by disease such as App.

Low dose endotoxemia is caused by several health conditions including smoking, alcohol abuse, high fat diets, and aging. Several studies have correlated low dose endotoxemia with increased risks of atherosclerosis, diabetes, and Parkinson's disease. Unlike high doses of endotoxin which induce a strong but transient induction of pro-inflammatory mediators, low doses of endotoxin result in a mild but chronic induction of pro-inflammatory genes. The central hypothesis of our study was that if low doses of endotoxin are capable of inducing mild prolonged inflammation, then a unique signaling circuit must be utilized. In the first study, the molecular mechanisms for the persistent induction of lipocalin 2 (LCN2) in response to 100 ng/mL of lipopolysaccharide (LPS) in kidney fibroblasts was examined. It appears that the intracellular signaling network responsible for the persistent induction of LCN2 requires both activator protein-1 (AP-1) and CCAAT/enhancer binding protein delta (C/ebpδ). Interleukin-1 receptor-associated kinase 1 (IRAK-1) is critical for LCN2 expression. In the second study, the molecular mechanisms governing the persistent induction of interleukin 6 (IL-6) upon a 50 pg/mL challenge of LPS in macrophages was examined. At this dose, only the persistent activation of cJun N-terminal kinase (JNK) and C/ebpδ was observed. IL-6 transcription requires the transient recruitment of activating transcription factor 2 (ATF2) and the persistent recruitment of C/ebpδ to the IL-6 promoter. In the third study, the molecular mechanisms that mediate LPS-induced priming was examined. The results demonstrate that macrophages are able to sense their prior history of exposure to LPS that result in either a priming or tolerance phenotype upon a secondary challenge of LPS. Results suggest that this sensing mechanism involves cross-talk between IRAK-1 and phosphoinositide-3-kinase (PI3K). Collectively, these studies indicate that JNK and C/ebpδ are the primary players responsible for the persistent expression of pro-inflammatory genes during low dose endotoxemia. IRAK-1 is a key intracellular signaling kinase that mediates signaling at low doses of LPS. IRAK-1 is not only critical for low dose induced expression, but also for LPS-induced priming. This research has revealed a novel signaling pathway that could provide new molecular targets for drug development against chronic inflammatory diseases. / Ph. D.

The overall aim of this thesis was to establish strategies to minimize secondary damage to the injured spinal cord. Secondary damage that follows spinal cord injury (SCI) involves inflammatory and excitotoxic pathways. Regulation of these pathways using immunomodulatory and neuroprotective substances potentially protects the injured spinal cord from further damage. We also developed and studied resorbable biomaterials to be used as carriers for potential neuroprotectants to the injured spinal cord. We used transversal spinal cord slice cultures (SCSCs) derived from postnatal mice as a model. SCSCs were maintained on different biomaterials and were studied after treatment with immunomodulatory and/or neurotrophic factors. They were further excitotoxically injured and subsequently treated with interleukin-1 receptor antagonist (IL1RA) or by neural crest stem cell (NCSC)-transplantation. The results show that biocompatible and resorbable hydrogels based on hyaluronic acid (HA) preserved neurons in SCSCs to a much higher extent than a conventional collagen-based biomaterial or standard polyethylene terephthalate (PET) membrane inserts. Glial activation was limited in the cultures maintained on HA-based hydrogel. The anti-inflammatory factor IL1RA protected SCSCs from degenerative mechanisms that occur during in vitro incubation, and IL1RA also protected SCSCs from excitotoxic injury induced by N-Methyl-d-Aspartate (NMDA). IL1RA specifically protected neurons that resided in the ventral horn, while other neuronal populations such as dorsal horn neurons and Renshaw cells did not respond to treatment. Finally, transplantation of NCSCs onto excitotoxically injured SCSCs protected from neuronal loss, apoptosis and glial activation, while NCSCs remained undifferentiated. The results presented in this thesis indicate that carriers based on HA seem to be more suitable than conventional collagen-based biomaterials since they enhance neuronal survival per se. The observed neuroprotection is likely due to biomechanical properties of HA. IL1RA protects SCSCs from spontaneous degeneration and from NMDA-induced injury, suggesting that excitotoxic mechanisms can be modulated through anti-inflammatory pathways. Different neuronal populations are affected by IL1RA to various degrees, suggesting that a combination of different neuroprotectants should be used in treatment strategies after SCI. Finally, NCSCs seem to protect SCSCs from excitotoxic injury through paracrine actions, since they remain undifferentiated and do not migrate into the tissue during in vitro incubation. It seems that combinations of neuroprotectants and carrier substances should be considered rather than one single strategy when designing future treatments for SCI. Incorporation of neuroprotectants such as IL1RA combined with stem cells in injectable biocompatible carriers based on HA is the final goal of our group in the treatment of SCI.