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Current Search:
'Biochemistry' in all fields;
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Displaying Results 76 - 100 of 301 on page 4 of 13
Marked
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Increased HLA-DR and decreased CD3 on human intestinal intraepithelial lymphocytes: evidence of activation?
(1996)
KELLEHER, DERMOT P; O'FARRELLY, CLIONA
Increased HLA-DR and decreased CD3 on human intestinal intraepithelial lymphocytes: evidence of activation?
(1996)
KELLEHER, DERMOT P; O'FARRELLY, CLIONA
Abstract:
peer-reviewed
BACKGROUND: Activation of circulating T lymphocytes results in expression of HLA-DR, interleukin-2 receptor (IL-2R), transferrin receptor (TrR), and decreased amounts of surface CD3. AIM: To examine the activation status of human intestinal intraepithelial lymphocytes (IELs) by flow cytometry. PATIENTS AND METHODS: Duodenal biopsy specimens from 12 patients provided a source of intestinal lymphocytes. Dual colour analysis was used to compare activation marker expression by IELs with peripheral blood (PB) T lymphocytes from the same patients. RESULTS: All human IEL populations express HLA-DR and their density of expression is lower than on the small population of HLA-DR+ resting PB T lymphocytes (mean fluorescence intensity (MFI) 52.9, range 19.8-94.8 v 152.6 range, 49.1-320.3; p < 0.01). The density of CD3 was significantly reduced on IELs (MFI 465.8, range 228.7-660), compared with PB T lymphocytes (756.3, range 444.5-1573.7; p < 0.009). IL-2R was not detect...
http://hdl.handle.net/2262/34192
Marked
Mark
The thioredoxin system of Helicobacter pylori
(2000)
KELLEHER, DERMOT P
The thioredoxin system of Helicobacter pylori
(2000)
KELLEHER, DERMOT P
Abstract:
peer-reviewed
This paper describes the purification of thioredoxin reductase (TR) and the characterization, purification, and cloning of thioredoxin (Trx) from Helicobacter pylori. Purification, amino acid sequence analysis, and molecular cloning of the gene encoding thioredoxin revealed that it is a 12-kDa protein which possesses the conserved redox active motif CGPC. The gene encoding Trx was amplified by polymerase chain reaction and inserted into a pET expression vector and used to transformEscherichia coli. Trx was overexpressed by induction with isopropyl-1-thio-β-d-galactopyranoside as a decahistidine fusion protein and was recovered from the cytoplasm as a soluble and active protein. The redox activity of this protein was characterized using several mammalian proteins of different architecture but all containing disulfide bonds. H. pylorithioredoxin efficiently reduced insulin, human immunoglobulins (IgG/IgA/sIgA), and soluble mucin. Subcellular fractionation analysis of...
http://hdl.handle.net/2262/34086
Marked
Mark
Interleukin-1 activates transcription factor NF kappa B in glial cells
(1993)
O'NEILL, LUKE ANTHONY JOHN
Interleukin-1 activates transcription factor NF kappa B in glial cells
(1993)
O'NEILL, LUKE ANTHONY JOHN
Abstract:
peer-reviewed
Recombinant human interleukin-1 (IL-1) alpha and beta were found to activate a latent cytosolic form of the transcription factor NF kappa B in rat C6 glioma. IL-1 beta was 10 times more potent than IL-1 alpha for this activity and both were inhibited by the IL-1 receptor antagonist. The activation was detectable from 20 min and remained sustained for up to 24 h. The electrophoretic mobility of the activated complex was shown to be different from that of the corresponding complexes in another IL-1-responsive cell line, the murine thymoma line EL4.NOB-1. C6 cells, when transiently transfected with five NF kappa B consensus sequence repeats linked to the reporter gene chloramphenicol acetyltransferase (CAT), demonstrated increased CAT activity in response to IL-1 beta treatment. The activation of NF kappa B in glial cells may thus represent an early step in IL-1 signalling in brain and is likely to have consequences for IL-1-induced gene expression in these cells.
http://hdl.handle.net/2262/33730
Marked
Mark
2-mercaptoethanol restores the ability of nuclear factor kappa B (NF kappa B) to bind DNA in nuclear extracts from interleukin 1-treated cells incubated with pyrollidine dithiocarbamate (PDTC)
(1997)
O'NEILL, LUKE ANTHONY JOHN
2-mercaptoethanol restores the ability of nuclear factor kappa B (NF kappa B) to bind DNA in nuclear extracts from interleukin 1-treated cells incubated with pyrollidine dithiocarbamate (PDTC)
(1997)
O'NEILL, LUKE ANTHONY JOHN
Abstract:
The metal chelator and anti-oxidant pyrollidine dithiocarbamate (PDTC) has been used extensively in studies implicating reactive oxygen intermediates in the activation of nuclear factor kappa B (NF kappa B). In agreement with other studies, we have shown that PDTC inhibits NF kappa B activation in response to the pro-inflammatory cytokines interleukin 1 (IL1) and tumour necrosis factor (TNF). However, we have found that the inhibition was reversed by treatment of inhibited nuclear extracts with the reducing agent 2-mercaptoethanol. This was observed in extracts prepared from IL1-treated EL4.NOB-1 thymoma cells and TNF-treated Jurkat E6.1 lymphoma cells. These results suggested that the inhibition was caused by oxidation of NF kappa B on a sensitive thiol, possibly on the p50 subunit (which was detected in NF kappa B complexes in both cell types), and not by inhibition of the activation pathway. The possibility that PDTC was acting as a pro-oxidant was therefore investigated. PDTC ca...
http://hdl.handle.net/2262/33734
Marked
Mark
Staurosporine, but not Ro 31-8220, induces interleukin 2 production and synergizes with interleukin 1alpha in EL4 thymoma cells.
(1997)
O'NEILL, LUKE ANTHONY JOHN
Staurosporine, but not Ro 31-8220, induces interleukin 2 production and synergizes with interleukin 1alpha in EL4 thymoma cells.
(1997)
O'NEILL, LUKE ANTHONY JOHN
Abstract:
peer-reviewed
Protein kinase C (PKC) has been implicated in interleukin 1 (IL1) signal transduction in a number of cellular systems, either as a key event in IL1 action or as a negative regulator. Here we have examined the effects of two PKC inhibitors, staurosporine and the more selective agent Ro 31-8220, on IL1 responses in the murine thymoma line EL4.NOB-1. A 1 h pulse of staurosporine was found to strongly potentiate the induction of IL2 by IL1alpha in these cells. In contrast, neither a pulse nor prolonged incubation with Ro 31-8220 affected the response to IL1alpha. Both agents blocked the response to PMA, however. A 1 h pulse of staurosporine was also found to induce IL2 production on its own, activate the transcription factor nuclear factor kappaB (NFkappaB) and increase the expression of a NFkappaB-linked reporter gene. It synergized with IL1alpha in all of these responses. Ro 31-8220 was again without effect, although both staurosporine and Ro 31-8220 blocked the acti...
http://hdl.handle.net/2262/33726
Marked
Mark
Signal transduction pathways activated by the IL-1 receptor family: ancient signaling machinery in mammals, insects, and plants.
(1998)
O'NEILL, LUKE ANTHONY JOHN
Signal transduction pathways activated by the IL-1 receptor family: ancient signaling machinery in mammals, insects, and plants.
(1998)
O'NEILL, LUKE ANTHONY JOHN
Abstract:
peer-reviewed
Interleukin-1 (IL-1) is a central regulator of the immune and inflammatory responses. Recently, significant advances have been made in the area of IL-1 receptors and IL-1 signal transduction. A family of proteins has been described that share significant homology in their signaling domains with the Type I IL-1 receptor (IL-1RI). These include the IL-1 receptor accessory protein (IL-1AcP), which does not bind IL-1 but is essential for IL-1 signaling; a Drosophila protein Toll; a number of human Toll-like receptors (hTLRs); the putative IL-18/IL-1-gamma receptor IL-1Rrp (IL-1 receptor-related protein); and a number of plant proteins. All appear to be involved in host responses to injury and infection. These homologies also extend to novel signaling proteins implicated in IL-1 action. Two IL-1 receptor-associated kinases, IRAK-1 and IRAK-2, which have homologs in Drosophila (Pelle) and plants (Pto), have been implicated in the activation of the transcription factor, n...
http://hdl.handle.net/2262/33709
Marked
Mark
Characterization of CD44 induction by IL-1: a critical role for Egr-1
(1999)
O'NEILL, LUKE ANTHONY JOHN
Characterization of CD44 induction by IL-1: a critical role for Egr-1
(1999)
O'NEILL, LUKE ANTHONY JOHN
Abstract:
peer-reviewed
he adhesion molecule CD44 is a multifunctional, ubiquitously expressed glycoprotein that participates in the process of leukocyte recruitment to sites of inflammation and to their migration through lymphatic tissues. In this study, we have investigated the effect of the proinflammatory cytokine IL-1 on CD44 gene expression in the human immortalized endothelial cell line ECV304. Immunoblotting of cell extracts showed constitutive expression of a 85-kDa protein corresponding to the standard form of CD44, which was potently up-regulated following IL-1 treatment. Furthermore, IL-1 induced expression of v3- and v6-containing isoforms of CD44, which migrated at 110 and 140–180 kDa, respectively. The effect of IL-1 on CD44 standard, v3- and v6-containing isoforms was dose and time dependent and was inhibited in the presence of IL-1 receptor antagonist. To elucidate the molecular mechanisms regulating CD44 expression in response to IL-1, we investigated the effect of IL-1 ...
http://hdl.handle.net/2262/33707
Marked
Mark
The interleukin-1 receptor/Toll-like receptor superfamily: signal generators for pro-inflammatory interleukins and microbial products
(2000)
BOWIE, ANDREW; O'NEILL, LUKE ANTHONY JOHN
The interleukin-1 receptor/Toll-like receptor superfamily: signal generators for pro-inflammatory interleukins and microbial products
(2000)
BOWIE, ANDREW; O'NEILL, LUKE ANTHONY JOHN
Abstract:
peer-reviewed
The interleukin-1 (IL-1) receptor/Toll-like receptor (TLR) superfamily is a recently defined and expanding group of receptors that participate in host responses to injury and infection. The superfamily is defined by the Toll/IL-1 receptor (TIR) domain, which occurs in the cytosolic region of family members, and is further subdivided into two groups based on homology to either the Type I IL-1 receptor or Drosophila Toll receptor extracellular domain. The former group includes the receptor for the important Th1 cytokine IL-18, and T1/ST2, which may have a role in Th2 cell function. The latter group includes six mammalian TLRs, including TLR2 and TLR4, that largely mediate the host response to gram-positive and gram-negative bacteria, respectively. Whether bacterial products are actual ligands for TLRs, or whether they generate ligands via as yet unidentified pattern recognition receptors, has yet to be determined. Signaling pathways activated via the TIR domain trigg...
http://hdl.handle.net/2262/33703
Marked
Mark
A46R and A52R from vaccinia virus are antagonists of host IL-1 and toll-like receptor signaling
(2000)
BOWIE, ANDREW; O'NEILL, LUKE ANTHONY JOHN
A46R and A52R from vaccinia virus are antagonists of host IL-1 and toll-like receptor signaling
(2000)
BOWIE, ANDREW; O'NEILL, LUKE ANTHONY JOHN
Abstract:
peer-reviewed
Poxviruses employ many strategies to evade and neutralize the host immune response. In this study, we have identified two vaccinia virus ORFs, termed A46R and A52R, that share amino acid sequence similarity with the Toll/IL-1 receptor (TIR) domain, a motif that defines the IL-1/Toll-like receptor (TLR) superfamily of receptors, which have a key role in innate immunity and inflammation. When expressed in mammalian cells, the protein products of both ORFs were shown to interfere specifically with IL-1 signal transduction. A46R partially inhibited IL-1-mediated activation of the transcription factor NFkappaB, and A52R potently blocked both IL-1- and TLR4-mediated NFkappaB activation. MyD88 is a TIR domain-containing adapter molecule known to have a central role in both IL-1 and TLR4 signaling. A52R mimicked the dominant-negative effect of a truncated version of MyD88 on IL-1, TLR4, and IL-18 signaling but had no effect on MyD88-independent signaling pathways. Therefor...
http://hdl.handle.net/2262/33701
Marked
Mark
Vitamin C inhibits NF-kappa B activation by TNF via the activation of p38 mitogen-activated protein kinase
(2000)
BOWIE, ANDREW; O'NEILL, LUKE ANTHONY JOHN
Vitamin C inhibits NF-kappa B activation by TNF via the activation of p38 mitogen-activated protein kinase
(2000)
BOWIE, ANDREW; O'NEILL, LUKE ANTHONY JOHN
Abstract:
peer-reviewed
The transcription factor NF-kappaB is a central mediator of altered gene expression during inflammation, and is implicated in a number of pathologies, including cancer, atherosclerosis, and viral infection. We report in this study that vitamin C inhibits the activation of NF-kappaB by multiple stimuli, including IL-1 and TNF in the endothelial cell line ECV304 and in primary HUVECs. The induction of a NF-kappaB-dependent gene, IL-8, by TNF was also inhibited. The effect requires millimolar concentrations of vitamin C, which occur intracellularly in vivo, particularly during inflammation. Vitamin C was not toxic to cells, did not inhibit another inducible transcription factor, STAT1, and had no effect on the DNA binding of NF-kappaB. Inhibition by vitamin C was not simply an antioxidant effect, because redox-insensitive pathways to NF-kappaB were also blocked. Vitamin C was shown to block IL-1- and TNF-mediated degradation and phosphorylation of I-kappaBalpha (inhib...
http://hdl.handle.net/2262/33699
Marked
Mark
Protective levels of diphtheria-neutralizing antibody induced in healthy volunteers by unilateral priming-boosting intranasal immunization associated with restricted ipsilateral mucosal secretory immunoglobulin a
(2003)
MILLS, KINGSTON
Protective levels of diphtheria-neutralizing antibody induced in healthy volunteers by unilateral priming-boosting intranasal immunization associated with restricted ipsilateral mucosal secretory immunoglobulin a
(2003)
MILLS, KINGSTON
Abstract:
peer-reviewed
Subunit intranasal vaccines offer the prospect of inducing combined systemic-mucosal immunity against mucosally transmitted infections such as human immunodeficiency virus. However, although human studies have demonstrated the induction of active immunity, secretory immunoglobulin A (sIgA) responses are variable, and no study has demonstrated protection by accepted vaccine-licensing criteria as measured by direct toxin-neutralizing activity. Using the genetically inactivated mutant diphtheria toxoid CRM197 in a bioadhesive polycationic polysaccharide chitosan delivery system, we found that a single nasal immunization was well tolerated and boosted antitoxin neutralizing activity in healthy volunteers, which could be further boosted by a second immunization. The neutralizing activity far exceeded accepted protective levels and was equivalent to that induced by standard intramuscular vaccine and significantly greater than intranasal immunization with CRM197 in the ab...
http://hdl.handle.net/2262/33332
Marked
Mark
Transactivation by the p65 subunit of NF-kappaB in response to interleukin-1 (IL-1) involves MyD88, IL-1 receptor-associated kinase 1, TRAF-6, and Rac1
(2001)
BOWIE, ANDREW; O'NEILL, LUKE ANTHONY JOHN
Transactivation by the p65 subunit of NF-kappaB in response to interleukin-1 (IL-1) involves MyD88, IL-1 receptor-associated kinase 1, TRAF-6, and Rac1
(2001)
BOWIE, ANDREW; O'NEILL, LUKE ANTHONY JOHN
Abstract:
peer-reviewed
We have examined the involvement of components of the interleukin-1 (IL-1) signaling pathway in the transactivation of gene expression by the p65 subunit of NF-kappaB. Transient transfection of cells with plasmids encoding wild-type MyD88, IL-1 receptor-associated kinase 1 (IRAK-1), and TRAF-6 drove p65-mediated transactivation. In addition, dominant negative forms of MyD88, IRAK-1, and TRAF-6 inhibited the IL-1-induced response. In cells lacking MyD88 or IRAK-1, no effect of IL-1 was observed. Together, these results indicate that MyD88, IRAK-1, and TRAF-6 are important downstream regulators of IL-1-mediated p65 transactivation. We have previously shown that the low-molecular-weight G protein Rac1 is involved in this response. Constitutively active RacV12-mediated transactivation was not inhibited by dominant negative MyD88, while dominant negative RacN17 inhibited the MyD88-driven response, placing Rac1 downstream of MyD88 on this pathway. Dominant negative RacN1...
http://hdl.handle.net/2262/33698
Marked
Mark
The anti-inflammatory cytokine, interleukin (IL)-10, blocks the inhibatory effect of IL-1 beta on long term potentiation - a role for JNK
(2001)
KELLY, AINE; LYNCH, MARINA ANNETTA; LYNCH, AILEEN MARIA; O'NEILL, LUKE ANTHONY JOHN
The anti-inflammatory cytokine, interleukin (IL)-10, blocks the inhibatory effect of IL-1 beta on long term potentiation - a role for JNK
(2001)
KELLY, AINE; LYNCH, MARINA ANNETTA; LYNCH, AILEEN MARIA; O'NEILL, LUKE ANTHONY JOHN
Abstract:
peer-reviewed
Several effects of the proinflammatory cytokine, interleukin-1β (IL-1β), have been described in the central nervous system, and one area of the brain where marked changes have been reported is the hippocampus. Among these changes are an IL-1β-induced inhibition of long term potentiation (LTP) in perforant path-granule cell synapses and an attenuation of glutamate release in synaptosomes prepared from the hippocampus. Evidence suggests that, at least in circulating cells, the anti-inflammatory cytokine, IL-10, antagonizes certain effects of IL-1. We investigated the effect of IL-10 on IL-1β-induced inhibition of LTP and glutamate release. The evidence presented indicates that IL-1β stimulates the stress-activated protein kinase, c-Jun-activated protein kinase (JNK), and IL-1 receptor-associated kinase, which may explain its inhibitory effect on release and LTP, and that IL-10 reversed the IL-1β-induced stimulation of JNK activity and inhibition of release and LTP. W...
http://hdl.handle.net/2262/33696
Marked
Mark
Structural complementarity of Toll/interleukin-1 receptor domains in Toll-like receptors and the adaptors Mal and MyD88
(2003)
O'NEILL, LUKE ANTHONY JOHN; DUNNE, AISLING
Structural complementarity of Toll/interleukin-1 receptor domains in Toll-like receptors and the adaptors Mal and MyD88
(2003)
O'NEILL, LUKE ANTHONY JOHN; DUNNE, AISLING
Abstract:
The Toll/interleukin 1 receptor (TIR) domain is a region found in the cytoplasmic tails of members of the Toll-like receptor/interleukin-1 receptor superfamily. The domain is essential for signaling and is also found in the adaptor proteins Mal (MyD88 adaptor-like) and MyD88, which function to couple activation of the receptor to downstream signaling components. Experimental structures of two Toll/interleukin 1 receptor domains reveal a α–β-fold similar to that of the bacterial chemotaxis protein CheY, and other evidence suggests that the adaptors can make heterotypic interactions with both the receptors and themselves. Here we show that the purified TIR domains of Mal and MyD88 can form stable heterodimers and also that Mal homodimers and oligomers are dissociated in the presence of ATP. To identify structural features that may contribute to the formation of signaling complexes, we produced models of the TIR domains from human Toll-like receptor 4 (TLR4), Mal, and MyD88. We found t...
http://hdl.handle.net/2262/33692
Marked
Mark
Transcriptional regulation of the human TRIF (TIR domain-containing adaptor protein inducing interferon beta) gene.
(2004)
MCGETTRICK-DILLON, ANNE; O'NEILL, LUKE ANTHONY JOHN
Transcriptional regulation of the human TRIF (TIR domain-containing adaptor protein inducing interferon beta) gene.
(2004)
MCGETTRICK-DILLON, ANNE; O'NEILL, LUKE ANTHONY JOHN
Abstract:
peer-reviewed
TRIF [TIR (Toll/interleukin-1 receptor) domain-containing adaptor protein inducing interferon beta; also known as TICAM-1 (TIR-containing adaptor molecule-1)] is a key adaptor for TLR3 (Toll-like receptor 3)- and TLR4-mediated signalling. We have performed a detailed annotation of the human TRIF gene and fine analysis of the basal and inducible promoter elements lying 5' to the site of initiation of transcription. Human TRIF maps to chromosome 19p13.3 and is flanked upstream by TIP47, which encodes the mannose 6-phosphate receptor binding protein, and downstream by a gene encoding FEM1a, a human homologue of the Caenorhabditis elegans Feminisation-1 gene. Using promoter-reporter deletion constructs, we identified a distal region with the ability to negatively regulate basal transcription and a proximal region containing an Sp1 (stimulating protein 1) site that confers approx. 75% of basal transcriptional activity. TRIF expression can be induced by multiple sti...
http://hdl.handle.net/2262/33690
Marked
Mark
IRAK-4: a new drug target in inflammation, sepsis and autoimmunity
(2002)
O'NEILL, LUKE ANTHONY JOHN
IRAK-4: a new drug target in inflammation, sepsis and autoimmunity
(2002)
O'NEILL, LUKE ANTHONY JOHN
Abstract:
peer-reviewed
http://hdl.handle.net/2262/33689
Marked
Mark
Bruton's tyrosine kinase is involved in p65-mediated transactivation and phosphorylation of p65 on serine 536 during NF-kappa B activation by LPS
(2005)
DOYLE, SARAH LOUISE; O'NEILL, LUKE ANTHONY JOHN
Bruton's tyrosine kinase is involved in p65-mediated transactivation and phosphorylation of p65 on serine 536 during NF-kappa B activation by LPS
(2005)
DOYLE, SARAH LOUISE; O'NEILL, LUKE ANTHONY JOHN
Abstract:
Bruton's tyrosine kinase (Btk) has recently been shown to participate in the induction of nuclear factor κB (NFκB)-dependent gene expression by the lipopolysaccharide (LPS) receptor Toll-like receptor-4 (TLR4). In this study we have examined the mechanism whereby Btk participates in this response. Treatment of the murine monocytic cell line Raw264.7 with LFM-A13, a specific Btk inhibitor, blocked LPS-induced NFκB-dependent reporter gene expression but not IκBα degradation. Transient transfection of HEK293 cells with Btk had no effect on NFκB-dependent reporter gene expression but strongly promoted transactivation of a reporter gene by a p65-Gal4 fusion protein. IκBα degradation activated by LPS was intact in macrophages from X-linked immunodeficiency (Xid) mice, which contain inactive Btk. Transfection of cells with a dominant negative form of Btk (BtkK430R) inhibited LPS-driven p65 mediated transactivation. Additionally LFM-A13 impaired phosphorylation of serine 536 on p65 ind...
http://hdl.handle.net/2262/33447
Marked
Mark
The myristoylation of TRIF-related adaptor molecule is essential for Toll-like receptor 4 signal transduction
(2006)
MCGETTRICK-DILLON, ANNE; O'NEILL, LUKE ANTHONY JOHN
The myristoylation of TRIF-related adaptor molecule is essential for Toll-like receptor 4 signal transduction
(2006)
MCGETTRICK-DILLON, ANNE; O'NEILL, LUKE ANTHONY JOHN
Abstract:
peer-reviewed
TRIF-related adaptor molecule (TRAM) is the fourth Toll/IL-1 resistance domain-containing adaptor to be described that participates in Toll-like receptor (TLR) signaling. TRAM functions exclusively in the TLR4 pathway. Here we show by confocal microscopy that TRAM is localized in the plasma membrane and the Golgi apparatus, where it colocalizes with TLR4. Membrane localization of TRAM is the result of myristoylation because mutation of a predicted myristoylation site in TRAM (TRAM-G2A) brought about dissociation of TRAM from the membrane and its relocation to the cytosol. Further, TRAM, but not TRAM-G2A, was radiolabeled with [3H]myristate in vivo. Unlike wild-type TRAM, overexpression of TRAM-G2A failed to elicit either IFN regulatory factor 3 or NF-kappaB signaling. Moreover, TRAM-G2A was unable to reconstitute LPS responses in bone marrow-derived macrophages from TRAM-deficient mice. These observations provide clear evidence that the myristoylation of TRAM targe...
http://hdl.handle.net/2262/33444
Marked
Mark
MyD88 adapter-like (Mal) is phosphorylated by Bruton's tyrosine kinase during TLR2 and TLR4 signal transduction
(2006)
O'NEILL, LUKE ANTHONY JOHN; DUNNE, AISLING
MyD88 adapter-like (Mal) is phosphorylated by Bruton's tyrosine kinase during TLR2 and TLR4 signal transduction
(2006)
O'NEILL, LUKE ANTHONY JOHN; DUNNE, AISLING
Abstract:
Members of the Toll-like receptor (TLR) family are essential players in activating the host innate immune response against infectious microorganisms. All TLRs signal through Toll/interleukin 1 receptor domain-containing adapter proteins. MyD88 adapter-like (Mal) is one such adapter that specifically is involved in TLR2 and TLR4 signaling. When overexpressed we have found that Mal undergoes tyrosine phosphorylation. Three possible phospho-accepting tyrosines were identified at positions 86, 106, and 187, and two mutant forms of Mal in which tyrosines 86 and 187 were mutated to phenylalanine acted as dominant negative inhibitors of NF-kappaB activation by lipopolysaccharide (LPS). Activation of THP-1 monocytic cells with the TLR4 agonist LPS and the TLR2 agonist macrophage-activating lipopeptide-2 induced phosphorylation of Mal on tyrosine residues. We found that the Bruton's tyrosine kinase (Btk) inhibitor LFM-A13 could block the endogenous phosphorylation of Mal on tyrosine in ...
http://hdl.handle.net/2262/33445
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Trif-related adapter molecule is phosphorylated by protein kinase C epsilon during Toll-like receptor 4 signalling
(2006)
MCGETTRICK-DILLON, ANNE; O'NEILL, LUKE ANTHONY JOHN; PALSSON, EVA
Trif-related adapter molecule is phosphorylated by protein kinase C epsilon during Toll-like receptor 4 signalling
(2006)
MCGETTRICK-DILLON, ANNE; O'NEILL, LUKE ANTHONY JOHN; PALSSON, EVA
Abstract:
peer-reviewed
PKCε has been shown to play a key role in the effect of the Gram-negative bacterial product LPS; however, the target for PKCε in LPS signaling is unknown. LPS signaling is mediated by Toll-like receptor 4, which uses four adapter proteins, MyD88, MyD88 adapter-like (Mal), Toll/IL-1R domain-containing adapter inducing IFN-β (Trif), and Trif-related adapter molecule (TRAM). Here we show that TRAM is transiently phosphorylated by PKCε on serine-16 in an LPS-dependent manner. Activation of IFN regulatory factor 3 and induction of the chemokine RANTES, which are both TRAM-dependent, were attenuated in PKCε-deficient cells. TRAMS16A is inactive when overexpressed and is attenuated in its ability to reconstitute signaling in TRAM-deficient cells. We have therefore uncovered a key process in Toll-like receptor 4 signaling, identifying TRAM as the target for PKCε.
http://hdl.handle.net/2262/33442
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A functional variant in MAL/TIRAP and protection against invasive pneumococcal disease, bacteraemia, malaria and tuberculosis
(2007)
O'NEILL, LUKE ANTHONY JOHN; DUNNE, AISLING
A functional variant in MAL/TIRAP and protection against invasive pneumococcal disease, bacteraemia, malaria and tuberculosis
(2007)
O'NEILL, LUKE ANTHONY JOHN; DUNNE, AISLING
Abstract:
Toll-like receptors (TLRs) and members of their signaling pathway are important in the initiation of the innate immune response to a wide variety of pathogens1, 2, 3. The adaptor protein Mal (also known as TIRAP), encoded by TIRAP (MIM 606252), mediates downstream signaling of TLR2 and TLR4 (refs. 4–6). We report a case-control study of 6,106 individuals from the UK, Vietnam and several African countries with invasive pneumococcal disease, bacteremia, malaria and tuberculosis. We genotyped 33 SNPs, including rs8177374, which encodes a leucine substitution at Ser180 of Mal. We found that heterozygous carriage of this variant associated independently with all four infectious diseases in the different study populations. Combining the study groups, we found substantial support for a protective effect of S180L heterozygosity against these infectious diseases (N = 6,106; overall P = 9.6 10-8). We found that the Mal S180L variant attenuated TLR2 signal transduction.
http://hdl.handle.net/2262/33406
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Camelpox virus encodes a schlafen-like protein that affects orthopoxvirus
(2007)
O'NEILL, LUKE ANTHONY JOHN
Camelpox virus encodes a schlafen-like protein that affects orthopoxvirus
(2007)
O'NEILL, LUKE ANTHONY JOHN
Abstract:
peer-reviewed
Camelpox virus (CMLV) gene 176R encodes a protein with sequence similarity to murine schlafen (m-slfn) proteins. In vivo, short and long members of the m-slfn family inhibited T-cell development, whereas in vitro, only short m-slfns caused arrest of fibroblast growth. CMLV 176 protein (v-slfn) is most closely related to short m-slfns; however, when expressed stably in mammalian cells, v-slfn did not inhibit cell growth. v-slfn is a predominantly cytoplasmic 57 kDa protein that is expressed throughout infection. Several other orthopoxviruses encode v-slfn proteins, but the v-slfn gene is fragmented in all sequenced variola virus and vaccinia virus (VACV) strains. Consistent with this, all 16 VACV strains tested do not express a v-slfn detected by polyclonal serum raised against the CMLV protein. In the absence of a small animal model to study CMLV pathogenesis, the contribution of CMLV v-slfn to orthopoxvirus virulence was studied via its expression in an attenuated...
http://hdl.handle.net/2262/33437
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Signaling by Toll-like receptors 8 and 9 requires Bruton's tyrosine kinase.
(2007)
FEIGHERY, CONLETH FRANCIS; DOYLE, SARAH LOUISE; O'NEILL, LUKE ANTHONY JOHN
Signaling by Toll-like receptors 8 and 9 requires Bruton's tyrosine kinase.
(2007)
FEIGHERY, CONLETH FRANCIS; DOYLE, SARAH LOUISE; O'NEILL, LUKE ANTHONY JOHN
Abstract:
Toll-like receptors (TLRs) are a primary surveillance system for the detection of pathogens and are crucial to the activation of host defense. TLR7 and TLR8 sense single-stranded RNA from viruses or host ribonucleoproteins and synthetic imidazoquinolines such as R848, whereas TLR9 senses unmethylated CpG motifs in viral and bacterial DNA and in host DNA. Here we report the endogenous interaction between Brutons's tyrosine kinase (Btk) and human TLR8 and TLR9 in the monocytic cell line THP1. We also show that R848, single-stranded RNA, and CpGB-DNA activate Btk in THP1 cells as shown by phosphorylation of the tyrosine 223 residue of Btk and also by increased autokinase activity. We demonstrate that Btk is required for NFkappaB activation, participating in the pathway to increased phosphorylation of p65 on serine 536 activated by TLR8 and TLR9. Finally we demonstrate that peripheral blood mononuclear cells from patients with X-linked agammaglobulinaemia (XLA) that have dysfunctio...
http://hdl.handle.net/2262/33433
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Glucocorticoids inhibit IRF3 phosphorylation in response to Toll-like
(2008)
O'NEILL, LUKE ANTHONY JOHN; CARPENTER, SUSAN; PALSSON MCDERMOTT, EVA
Glucocorticoids inhibit IRF3 phosphorylation in response to Toll-like
(2008)
O'NEILL, LUKE ANTHONY JOHN; CARPENTER, SUSAN; PALSSON MCDERMOTT, EVA
Abstract:
Phosphorylation of the transcription factor interferon regulatory factor 3 (IRF3) is essential for the induction of promoters which contain the interferon-stimulated response element (ISRE). IRF3 can be activated by Toll-like receptor 3 (TLR3) in response to the double-stranded RNA mimic poly(I-C) and by TLR4 in response to lipopolysaccharide (LPS). Here we have analyzed the effect of the glucocorticoid dexamethasone on this response. Dexamethasone inhibited the induction of the ISRE-dependent gene RANTES (regulated on activation normal T cell expressed and secreted) in both U373-CD14 cells and human peripheral blood mononuclear cells and also an ISRE luciferase construct, activated by either TLR3 or TLR4. It also inhibited increased phosphorylation of IRF3 in its N terminus in response to LPS and in its C terminus on Ser-396 in response to either poly(I-C) or LPS. Several dexamethasone-induced phosphatases were tested for possible involvement in these effects; MKP1 did not appear t...
http://hdl.handle.net/2262/33430
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T cell subpopulations in B cell chronic lymphocytic leukemias
(1984)
MILLS, KINGSTON
T cell subpopulations in B cell chronic lymphocytic leukemias
(1984)
MILLS, KINGSTON
Abstract:
peer-reviewed
http://hdl.handle.net/2262/33386
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