Regulation of Complement Activation at Sites of Chronic Inflammation

Guc, Dicle (1993) Regulation of Complement Activation at Sites of Chronic Inflammation. PhD thesis, University of Glasgow.

Abstract

Biosynthesis of the complement regulatory proteins Cl-INH, C4-bp, factor H, factor I, S protein, SP-40,40, DAF, MCP, CD59 and CRl has been investigated in synovial membrane tissue cultures obtained from normal individuals, and patients with osteoarthritis (OA) and rheumatoid arthritis (RA) by a combination of immunological and molecular biological techniques. Normal, OA and RA synovial membranes were capable of s5mthesizing this battery of regulatory complement proteins. The abundances of the mRNAs encoding the classical and alternative pathway secreted proteins were elevated in RA synovium as was that of CD59 mRNA. However, tissue samples were small and they may not reflect the whole situation in RA joints. The cells present in synovial membrane, endothelial cells, monocytes, synovial fibroblasts and peripheral blood lymphocytes, were cultured and investigated in order to find which cell types provide which regulatory proteins detected in synovial membrane. Cell-specific differences were observed in the regulatory complement proteins synthesized. ELISA, immunohistochemical staining. Northern blot and doubling-dilution dot-blot analysis showed that both monocytes and fibroblasts expressed nine, endothelial cells expressed eight and lymphocytes expressed six of the eleven components under standard culture conditions. All of the cell types investigated synthesized factor H, DAF, MCP, CD59 and MIP. Endothelial cells and fibroblasts did not synthesize C4-bp and CRl. Also cell-specific differences were observed in the amounts of secretion for some proteins. Furthermore, cell specific differences were, observed for the expression of different mRNA species of DAF, MCP and CD59 between endothelial cells and fibroblasts. The effect of a cytokine, interferon-gamma (IFNgamma), on the synthesis of these complement regulatory proteins was also investigated. Treatment of endothelial cells with IFN? caused an increase of transcription rate of DAF, MCP and CD59 but this did not correlate with the protein levels on the cell surface detected by flow cytometry. This study indicated that complement regulatory protein s3mthesis is constitutive and varies between the different types of cells in synovial membrane. To some extent, the amounts of some complement regulatory proteins is regulated by IFN?.

• Item Type: Thesis (PhD)
• Qualification Level: Doctoral
• Additional Information: Adviser: George D Birnie
• Keywords: Immunology, Molecular biology
• Date of Award: 1993
• Depositing User: Enlighten Team
• Unique ID: glathesis:1993-74989
• Copyright: Copyright of this thesis is held by the author.
• Date Deposited: 27 Sep 2019 14:44
• Last Modified: 27 Sep 2019 14:44
• URI: https://theses.gla.ac.uk/id/eprint/74989