Use this URL to cite or link to this record in EThOS: https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.723770
Title: Immune regulation induced by apoptotic cells in health and in systemic lupus erythematosus (SLE)
Author: Simpson, Joanne Elizabeth
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 2016
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Abstract:
Systemic lupus erythematosus (SLE) is a chronic autoimmune disease where failure to remove apoptotic cells, due to a defect in phagocytic cells, or deficient opsonisation, leads to secondary necrosis and the release of DNA and chromatin. The nuclear constituents from apoptotic cells are targeted by autoantibodies, which form immune complexes. Immune complex-mediated TLR9 activation of plasmacytoid dendritic cells (pDCs) and subsequent secretion of interferon (IFN-α) is thought to drive inflammation in SLE. It is currently believed that pDCs do not normally respond to apoptotic cells, as self-DNA is hidden from TLR9. However, DNA and chromatin expressed on membrane bound apoptotic bodies is essential for inducing IL-10 secreting regulatory B cells through TLR9 stimulation. The overall objective of this thesis was to understand how apoptotic cells influence immune responses in health and in patients with SLE. Splenic mouse pDCs were activated with the synthetic TLR7 agonist R848 and TLR9 agonists CpGB and CpGA and were co-cultured with apoptotic cells, or with freeze-thawed necrotic cells. PDCs co-cultured with apoptotic cells down-regulated the expression of CD40 and CD86. When pDCs were activated by R848 or CpGB, IL-10, IFN-γ and IL-6 secretion was significantly induced in the presence of apoptotic cells. PDCs so cultured induced T cells to secrete immune-regulatory IL- 10. In contrast, co-culturing apoptotic cells with pDCs activated by CpGA, augmented IFN-α secretion. These cytokine responses by pDCs were only stimulated by DNA on whole apoptotic cells; not by free nucleic acids derived from necrotic cells. This data demonstrates that the inflammatory context in which pDCs sense whole apoptotic cells is crucial to determining the threshold of tolerance to apoptotic self. It questions the perception that pDCs see all apoptotic cells and their necrotic cellular debris as dangerous and suggests that there may be something intrinsically different about SLE apoptotic cells, which causes inflammation. SNPs near ATG5, a protein of the cell survival pathway autophagy, have been linked to SLE susceptibility, but the role of autophagy in SLE pathogenesis is unclear. We hypothesised that dysfunctional autophagy is linked to abnormal apoptosis of SLE lymphocytes. Western blotting revealed that ATG5-ATG12 protein complex expression was significantly reduced in SLE lymphocytes and they failed to convert LC3-I to LC3- II, the hallmark of a functioning autophagy pathway, which caused accelerated secondary necrosis. Apoptotic SLE lymphocytes had an impaired ability to stimulate IL-10 secreting regulatory B cells and they induced pro-inflammatory cytokine secretion by monocyte-derived macrophages. Phagocytosis of apoptotic SLE lymphocytes by healthy macrophages was also impaired; however this was independent of ATG5 protein expression. The novel findings of this thesis suggest SLE apoptotic lymphocytes are intrinsically pro-inflammatory, which may be caused by diminished autophagy leading to an inability of lymphocytes to correctly execute apoptosis. Furthermore, inefficient clearance of SLE apoptotic cells results from a defect in the apoptotic cell, rather than the phagocytic cell.
Supervisor: Gray, Mohini Sponsor: Medical Research Council (MRC)
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID: uk.bl.ethos.723770  DOI: Not available
Keywords: apoptosis ; autoimmunity ; lymphacyte ; plasmacytoid dendritic cells ; pDCs ; systemic lupus erythematosus ; SLE ; tolerance
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