Rysnik O.,University of Oxford |
McHugh K.,University of Oxford |
van Duivenvoorde L.,Amsterdam Rheumatology and Immunology Center |
van Tok M.,Amsterdam Rheumatology and Immunology Center |
And 6 more authors.
Journal of Autoimmunity | Year: 2016
Objectives: Human leukocyte antigen (HLA)-B27 (B27) is the strongest genetic factor associated with development of Ankylosing Spondylitis and other spondyloarthropathies (SpA), yet the role it plays in disease pathogenesis remains unclear. We investigated the expression of potentially pathogenic non-conventional heavy chain forms (NC) of B27 in synovial and intestinal tissues obtained from SpA patients. We also determined the presence of NC-B27 in joints, lymphoid and gastrointestinal tissue from B27 transgenic (TG1) rats with M.tuberculosis-induced SpA. Methods: Expression of NC-B27 in human SpA joints and gut and in (21-3 × 283-2)F1 HLA-B27/Huβ2m rat tissue was determined by immunohistochemistry, flow cytometry and confocal microscopy analysis using HC10 and HD6 antibodies. Results: Both HC10- and HD6-reactive HLA molecules were present in synovial tissue from SpA patients. Both NC-B27 and KIR3DL2, a ligand for NC-B27, were expressed in inflamed terminal ileal tissues in patients with early SpA. Infiltrating cells in inflamed joint tissues isolated from B27 TG1 rats expressed high levels of NC-B27. NC-B27 were also expressed in joint-resident cells from ankle and tail joints of B27 TG1 rats prior to clinical arthritis. The expression of NC-B27 on B27 TG1 rat CD11b/c+, CD8α+, cells from spleens and LNs increased with animal age and disease progression. Conclusions: Non-conventional HLA class 1 molecules are expressed on resident and infiltrating cells in both synovial and GI tissues in human SpA. NC-B27 expression in joints and lymphoid tissues from B27 TG1 rats prior to the onset of arthritis is consistent with the hypothesis that they play a pathogenic role in SpA. © 2016 The Authors.
Humby F.,Queen Mary, University of London |
Kelly S.,Queen Mary, University of London |
Bugatti S.,University of Pavia |
Manzo A.,University of Pavia |
And 16 more authors.
Journal of Rheumatology | Year: 2016
Objective: Because limited data currently support the clinical utility of peripherally expressed biomarkers in guiding treatment decisions for patients with rheumatoid arthritis, the search has turned to the disease tissue. The strategic aim of the Outcome Measures in Rheumatology (OMERACT) synovitis working group over the years has been to develop novel diagnostic and prognostic synovial biomarkers. A critical step in this process is to refine and validate minimally invasive, technically simple, robust techniques to sample synovial tissue, for use both in clinical trials and routine clinical practice. The objective of the synovitis working group (SWG) at OMERACT 12 (2014) was to examine whether recently developed ultrasound (US)-guided synovial biopsy techniques could be validated according to the OMERACT filter for future clinical use recommendation. Methods: The SWG examined whether current data reporting US-guided synovial biopsy of both large and small joints addressed the OMERACT filters of truth, discrimination, and feasibility. Results: There are currently limited data examining the performance of US-guided synovial biopsy, mainly from observational studies. Thus, it remains critical to evaluate its performance, within the clinical trials context, against the current gold standard of arthroscopic biopsy, with particular reference to: (1) synovial tissue yield, (2) capacity to determine treatment response as measured by a validated synovial biomarker, and (3) tolerability of the procedure. Conclusion: We summarize the discrete work packages agreed to as requirements to validate US-guided synovial biopsy and therefore lead to a global consensus on the use of synovial biopsy for research and clinical practice. © 2016. All rights reserved.
Reynolds L.A.,University of Edinburgh |
Reynolds L.A.,University of British Columbia |
Harcus Y.,University of Edinburgh |
Smith K.A.,University of Edinburgh |
And 16 more authors.
Journal of Immunology | Year: 2014
Helminth parasites remain one of the most common causes of infections worldwide, yet little is still known about the immune signaling pathways that control their expulsion. C57BL/6 mice are chronically susceptible to infection with the gastrointestinal helminth parasite Heligmosomoides polygyrus. In this article, we report that C57BL/6 mice lacking the adapter protein MyD88, which mediates signaling by TLRs and IL-1 family members, showed enhanced immunity to H. polygyrus infection. Alongside increased parasite expulsion, MyD88-deficient mice showed heightened IL-4 and IL-17A production from mesenteric lymph node CD4+ cells. In addition, MyD88-/- mice developed substantial numbers of intestinal granulomas around the site of infection, which were not seen in MyD88-sufficient C57BL/6 mice, nor when signaling through the adapter protein TRIF (TIR domain-containing adapter-inducing IFN-β adapter protein) was also ablated. Mice deficient solely in TLR2, TLR4, TLR5, or TLR9 did not show enhanced parasite expulsion, suggesting that these TLRs signal redundantly to maintain H. polygyrus susceptibility in wild-type mice. To further investigate signaling pathways that are MyD88 dependent, we infected IL-1R1-/- mice with H. polygyrus. This genotype displayed heightened granuloma numbers compared with wild-type mice, but without increased parasite expulsion. Thus, the IL-1R-MyD88 pathway is implicated in inhibiting granuloma formation; however, protective immunity in MyD88-deficient mice appears to be granuloma independent. Like IL-1R1-/- and MyD88-/- mice, animals lacking signaling through the type 1 IFN receptor (i.e., IFNAR1-/-) also developed intestinal granulomas. Hence, IL-1R1, MyD88, and type 1 IFN receptor signaling may provide pathways to impede granuloma formation in vivo, but additional MyD88-mediated signals are associated with inhibition of protective immunity in susceptible C57BL/6 mice. Copyright © 2014 The Authors 0022-1767/14.
Orme R.P.,Keele University |
Orme R.P.,Cardiff Institute of Infection and Immunity |
Middleditch C.,Keele University |
Waite L.,Keele University |
Fricker R.A.,Keele University
Vitamins and Hormones | Year: 2015
Vitamin D has long been synonymous with bone health. More recently, new health benefits are continually being associated with vitamin D, including a burgeoning field on neuroprotective properties. This has generated a huge explosion of interest in recent years in the potential for vitamin D to be used not only as a therapeutic in neurodegenerative disease, including Parkinson's disease, but also as biomarkers and for risk association.With an emphasis on Parkinson's disease, this chapter will discuss recent evidence supporting the assertion that vitamin D can be a useful therapeutic agent used as an intervention therapy to be combined with existing treatments; and the case for further development of novel treatments utilizing the potential of vitamin D.In addition, we present novel, previously unpublished evidence showing that in a unilateral model of Parkinson's disease, vitamin D can not only reduce the extent of denervation, but that this is also reflected in functional benefit to the animals.The potential of vitamin D is slowly being realized; in the future, it will be widely associated with far more than just bone health and may even contribute to an elusive treatment of neurodegenerative illness. © 2015 Elsevier Inc.
Yap M.W.,UK National Institute for Medical Research |
Colbeck E.,UK National Institute for Medical Research |
Colbeck E.,Cardiff Institute of Infection and Immunity |
Ellis S.A.,UK National Institute for Medical Research |
And 3 more authors.
PLoS Pathogens | Year: 2014
Fv1 is the prototypic restriction factor that protects against infection by the murine leukemia virus (MLV). It was first identified in cells that were derived from laboratory mice and was found to be homologous to the gag gene of an endogenous retrovirus (ERV). To understand the evolution of the host restriction gene from its retroviral origins, Fv1s from wild mice were isolated and characterized. Most of these possess intact open reading frames but not all restricted N-, B-, NR-or NB-tropic MLVs, suggesting that other viruses could have played a role in the selection of the gene. The Fv1s from Mus spretus and Mus caroli were found to restrict equine infectious anemia virus (EIAV) and feline foamy virus (FFV) respectively, indicating that Fv1 could have a broader target range than previously thought, including activity against lentiviruses and spumaviruses. Analyses of the Fv1 sequences revealed a number of residues in the C-terminal region that had evolved under positive selection. Four of these selected residues were found to be involved in the novel restriction by mapping studies. These results strengthen the similarities between the two capsid binding restriction factors, Fv1 and TRIM5α, which support the hypothesis that Fv1 defended mice against waves of retroviral infection possibly including non-MLVs as well as MLVs. © 2014 Yap et al.
Mondhe M.,Northumbria University |
Mondhe M.,Cardiff Institute of Infection and Immunity |
Chessher A.,Northumbria University |
Goh S.,Royal Veterinary College University of London |
Stach J.E.M.,Northumbria University
PLoS ONE | Year: 2014
Broad-spectrum antimicrobials kill indiscriminately, a property that can lead to negative clinical consequences and an increase in the incidence of resistance. Species-specific antimicrobials that could selectively kill pathogenic bacteria without targeting other species in the microbiome could limit these problems. The pathogen genome presents an excellent target for the development of such antimicrobials. In this study we report the design and evaluation of species-selective peptide nucleic acid (PNA) antibacterials. Selective growth inhibition of B. subtilis, E. coli, K. pnuemoniae and S. enterica serovar Typhimurium in axenic or mixed culture could be achieved with PNAs that exploit species differences in the translation initiation region of essential genes. An S. Typhimurium-specific PNA targeting ftsZ resulted in elongated cells that were not observed in E. coli, providing phenotypic evidence of the selectivity of PNA-based antimicrobials. Analysis of the genomes of E. coli and S. Typhimurium gave a conservative estimate of >150 PNA targets that could potentially discriminate between these two closely related species. This work provides a basis for the development of a new class of antimicrobial with a tuneable spectrum of activity. © 2014 Mondhe et al.
Sherwani S.,Cardiff Institute of Infection and Immunity |
Blythe N.,Cardiff Institute of Infection and Immunity |
Farleigh L.,Cardiff Institute of Infection and Immunity |
Bugert J.J.,Cardiff Institute of Infection and Immunity
Methods in Molecular Biology | Year: 2012
Molluscum contagiosum virus (MCV), a poxvirus pathogenic for humans, replicates well in human skin in vivo, but not in vitro in standard monolayer cell cultures. In order to determine the nature of the replication deficiency in vitro, the MCV infection process in standard culture has to be studied step by step. The method described in this chapter uses luciferase and GFP reporter constructs to measure poxviral mRNA transcription activity in cells in standard culture infected with known quantities of MCV or vaccinia virus. Briefly, MCV isolated from human tissue specimen is quantitated by PCR and used to infect human HEK293 cells, selected for ease of transfection. The cells are subsequently transfected with a reporter plasmid encoding firefly luciferase gene under the control of a synthetic early/late poxviral promoter and a control plasmid encoding a renilla luciferase reporter under the control of a eukaryotic promoter. After 16 h, cells are harvested and tested for expression of luciferase. MCV genome units are quantitated by PCR targeting a genome area conserved between MCV and vaccinia virus. Using a GFP reporter plasmid, this method can be further used to infect a series of epithelial and fibroblast-type cell lines of human and animal origin to microscopically visualize MCV-infected cells, to assess late promoter activation, and, using these parameters, to optimize MCV infectivity and gene expression in more complex eukaryotic cell culture models. © 2012 Springer Science+Business Media, LLC.
Smith K.A.,University of Edinburgh |
Smith K.A.,Cardiff Institute of Infection and Immunity |
Maizels R.M.,University of Edinburgh
European Journal of Immunology | Year: 2014
IL-6 plays a pivotal role in favoring T-cell commitment toward a Th17 cell rather than Treg-cell phenotype, as established through in vitro model systems. We predicted that in the absence of IL-6, mice infected with the gastrointestinal helminth Heligmosomoides polygyrus would show reduced Th17-cell responses, but also enhanced Treg-cell activity and consequently greater susceptibility. Surprisingly, worm expulsion was markedly potentiated in IL-6-deficient mice, with significantly stronger adaptive Th2 responses in both IL-6-/- mice and BALB/c recipients of neutralizing anti-IL-6 monoclonal Ab. Although IL-6-deficient mice showed lower steady-state Th17-cell levels, IL-6-independent Th17-cell responses occurred during in vivo infection. We excluded the Th17 response as a factor in protection, as Ab neutralization did not modify immunity to H. polygyrus infection in BALB/c mice. Resistance did correlate with significant changes to the associated Treg-cell phenotype however, as IL-6-deficient mice displayed reduced expression of Foxp3, Helios, and GATA-3, and enhanced production of cytokines within the Treg-cell population. Administration of an anti-IL-2:IL-2 complex boosted Treg-cell proportions in vivo, reduced adaptive Th2 responses to WT levels, and fully restored susceptibility to H. polygyrus in IL-6-deficient mice. Thus, in vivo, IL-6 limits the Th2 response, modifies the Treg-cell phenotype, and promotes host susceptibility following helminth infection. © 2013 The Authors. European Journal of Immunology.
PubMed | Cardiff Institute of Infection and Immunity
Type: | Journal: Methods in molecular biology (Clifton, N.J.) | Year: 2012
Molluscum contagiosum virus (MCV), a poxvirus pathogenic for humans, replicates well in human skin in vivo, but not in vitro in standard monolayer cell cultures. In order to determine the nature of the replication deficiency in vitro, the MCV infection process in standard culture has to be studied step by step. The method described in this chapter uses luciferase and GFP reporter constructs to measure poxviral mRNA transcription activity in cells in standard culture infected with known quantities of MCV or vaccinia virus. Briefly, MCV isolated from human tissue specimen is quantitated by PCR and used to infect human HEK293 cells, selected for ease of transfection. The cells are subsequently transfected with a reporter plasmid encoding firefly luciferase gene under the control of a synthetic early/late poxviral promoter and a control plasmid encoding a renilla luciferase reporter under the control of a eukaryotic promoter. After 16 h, cells are harvested and tested for expression of luciferase. MCV genome units are quantitated by PCR targeting a genome area conserved between MCV and vaccinia virus. Using a GFP reporter plasmid, this method can be further used to infect a series of epithelial and fibroblast-type cell lines of human and animal origin to microscopically visualize MCV-infected cells, to assess late promoter activation, and, using these parameters, to optimize MCV infectivity and gene expression in more complex eukaryotic cell culture models.