Expression of the components and regulatory proteins of the alternative complement pathway and the membrane attack complex in normal and diseased synovium.
We have studied synthesis of the complement components and regulatory proteins of the alternative pathway and the membrane attack complex in synovial membrane. RNA was extracted from synovial tissue of patients with rheumatoid arthritis (RA) or osteoarthritis (OA) as well as from normal synovial membrane. Dot blot analysis showed the presence of mRNAs for all the complement components and regulatory proteins (C3, factor B, factor D, C5, C6, C7, C9, factor H, factor I, S-protein, SP-40, 40, DAF, MCP, CR1, CD59), except for properdin, C8 alpha, C8 beta and C8 gamma in all three types of synovial membrane studied. In an attempt to determine which components were synthesised by each cell type, monocytes (mononuclear phagocytes), human umbilical vein endothelial cells (HUVEC), synovial membrane fibroblasts (from normal, OA and RA synovial membrane) and peripheral blood lymphocytes were cultured in vitro and secretion rates of individual components were measured and total cellular RNA analysed by northern blotting. Monocytes secreted properdin, C3, and factor H but not factor B, factor I, C5, C6, C7, C8 or C9. Fibroblasts and endothelial cells secreted factor B, factor H and factor I, but not properdin, C5, C6, C7, C8 or C9. Lymphocytes did not secrete any of these components. mRNAs encoding C3, factor B, factor H, S-protein, SP-40, 40, MCP and DAF were detected in all three other cell types (monocytes, fibroblasts and HU-VEC), but factor I and CD59 mRNAs were not detected in monocytes. C5, C6, C7, C8 alpha, C8 beta, CD8 gamma and C9 mRNAs were not detected in any of the cell types studied.(ABSTRACT TRUNCATED AT 250 WORDS)
Guc
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Beatson Institute for Cancer Research, Glasgow, UK.
Gulati
P
P
Lemercier
C
C
Lappin
D
D
Birnie
G D
GD
Whaley
K
K
D001172:Arthritis, Rheumatoid / Q000378:metabolism* / Q000473:pathology / Q000503:physiopathology; D001483:Base Sequence; D015152:Blotting, Northern; D002478:Cells, Cultured; D003176:Complement C3 / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D003182:Complement C5 / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D003183:Complement C6 / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D003184:Complement C7 / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D003186:Complement C9 / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D017242:Complement Factor H / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D015938:Complement Membrane Attack Complex / Q000032:analysis* / Q000378:metabolism / Q000502:physiology; D003170:Complement Pathway, Alternative / Q000502:physiology*; D004730:Endothelium, Vascular / Q000737:chemistry / Q000166:cytology / Q000502:physiology; D005347:Fibroblasts / Q000737:chemistry / Q000166:cytology / Q000502:physiology; D006801:Humans; D007962:Leukocytes / Q000737:chemistry / Q000473:pathology / Q000502:physiology; D008562:Membrane Glycoproteins / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D009000:Monocytes / Q000737:chemistry / Q000473:pathology / Q000502:physiology; D015345:Oligonucleotide Probes; D010003:Osteoarthritis / Q000378:metabolism* / Q000473:pathology / Q000503:physiopathology; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D013583:Synovial Membrane / Q000737:chemistry* / Q000473:pathology / Q000502:physiology*; D019096:Vitronectin
D003176:Complement C3; D003182:Complement C5; D003183:Complement C6; D003184:Complement C7; D003186:Complement C9; D017242:Complement Factor H; D015938:Complement Membrane Attack Complex; D008562:Membrane Glycoproteins; D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D019096:Vitronectin; C493495:CFH protein, human