Investigating the role of Toll-like receptors in juvenile onset systemic lupus erythematosus



Thorbinson, Colin
Investigating the role of Toll-like receptors in juvenile onset systemic lupus erythematosus. [Unspecified]

[img] Microsoft Word (MPhil Thesis)
CT_MPhil_Corrected_Thesis_August_2009-2010.docx - Accepted Version
Access to this file is embargoed until Unspecified.
Available under License Creative Commons Attribution No Derivatives.

Download (1MB)
[img] PDF (Renamed and converted to pdf)
THORBINSONCol_Aug2010_1452.pdf
Access to this file is embargoed until Unspecified.
Available under License Creative Commons Attribution No Derivatives.

Download (2MB)
[img] PDF (Abridged version - converted to pdf)
ThorbinsonCol_Aug2010_1452(abridged_version).pdf - Accepted Version
Available under License Creative Commons Attribution No Derivatives.

Download (1MB)
[img] Microsoft Word (Abridged version)
ThorbinsonCol_Aug2010_1452(abridged_version).docx - Accepted Version
Access to this file is embargoed until Unspecified.
Available under License Creative Commons Attribution No Derivatives.

Download (819kB)

Abstract

Background: Juvenile-onset Systemic Lupus Erythematosus (JSLE) is a chronic debilitating multi-system autoimmune condition characterised by auto-antibody production directed against nuclear antigens. It is associated with a more severe onset and more aggressive clinical course than in adults, with poor prognostic factors presenting earlier in childhood. It has been proposed that dysregulated neutrophil apoptosis may be a source of autoantigen in JSLE. Toll-like receptors (TLRs) are essential in the function of the innate immune system recognising pathogenic material. Upon stimulation they initiate a non-specific immune response leading to the production of an antigen-specific immune defence and autoantibody production. TLRs have been implicated in the development of autoimmunity. TLRs 3, 7, 8 and 9 are capable of recognising nucleic autoantigens typical of SLE and their expression has been shown to positively correlate with anti-dsDNA titres and disease activity in adult-onset SLE. To date there have been no studies examining the role of TLRs in JSLE. Aim: To assess whether apoptotic neutrophils in JSLE are providing a source of nuclear autoantigen which are being detected through TLRs 3, 7, 8 and 9 resulting in an inflammatory response through activation of an adaptive autoimmune response. Methods: Peripheral Blood Mononuclear Cells (PBMCs) and B cells were isolated from JSLE patients, Juvenile Idiopathic Arthritis (JIA) (inflammatory controls) and non-inflammatory controls. TLR 3, 7-9 mRNA and protein expression was measured using quantitative PCR (qPCR) and flow cytometry respectively. PBMCs were incubated with TLR agonists, activation was measured by IFN-a protein and mRNA expression, by ELISA and qPCR respectively. Neutrophils were isolated from healthy controls and incubated with JSLE serum to induce apoptosis. Apoptotic neutrophils were incubated with PBMCs at varying concentrations for 6 hours. After which IFN-a mRNA expression was measured by qPCR. PBMCs were treated with a MyD88 inhibitor to block TLRs 7-9 or left untreated. MyD88 treated and untreated PBMCs were incubated with TLR9 agonist or apoptotic neutrophils for 6 hours. TLR activation was measured using IFN-a mRNA expression by qPCR. Results: JSLE patients have a significantly increased PBMC TLR 3, 8 and 9 protein and mRNA expression compared to controls (p

Item Type: Unspecified
Additional Information: Date: 2010-08 (completed)
Subjects: R Medicine > RC Internal medicine
R Medicine > RC Internal medicine > RC0254 Neoplasms. Tumors. Oncology (including Cancer)
Divisions: ?? sch_repro ??
Depositing User: Symplectic Admin
Date Deposited: 11 Jan 2011 10:58
Last Modified: 08 May 2020 16:28
URI: http://livrepository.liverpool.ac.uk/id/eprint/1452