Complement activation induces the expression of decay-accelerating factor on human mesangial cells

Toru Shibata, Fernando G Cosio, Daniel J. Birmingham

Research output: Contribution to journalArticle

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Abstract

In the present study we evaluated the effect of complement activation by immune complexes (IC) on the expression of decay-accelerating factor (DAF) on human mesangial cells (MC). MC in culture were incubated with an Ag (DNP-Gelatin) that binds to fibronectin present in the MC matrix. Subsequently, MC were incubated with anti-DNP antibodies in the presence of human serum. By immunoperoxidase staining we showed that these incubations resulted in IC formation and deposition of human C3 and terminal complement components (C5b-9) on the mesangial matrix and on the surface of MC. By immunoperoxidase staining and by RIA we showed that IC formation and complement activation significantly increased DAF expression on the MC plasma membrane. The induction of DAF expression was a consequence of deposition of terminal complement components on the MC because, zymosan-activated serum and IC formation in the presence of C5- or C8-deficient serum failed to increase MC DAF expression. Furthermore, the observed increased DAF expression was the consequence of increased DAF synthesis by MC. Thus, both cycloheximide and actinomycin D blocked the increase on MC DAF observed after incubation with IC and serum. MC DAF had biophysical and functional characteristics similar to DAF in other cells. Thus, 1) MC DAF was resistant to trypsin but was removed from the MC membrane by pronase; 2) phosphatidylinositol-specific phospholipase C removed 48 ± 4% of MC DAF indicating that MC DAF is anchored in the cell membrane by phosphatidylinositol groups; 3) DAF isolated from MC-inhibited complement-mediated hemolysis and demonstrated a molecular mass of 83 kDa. In conclusion, deposition of terminal complement components on human MC trigger new synthesis and membrane expression of DAF. Because DAF protects cells against complement-mediated lysis, we postulate that DAF may protect glomerular cells during IC and complement-mediated glomerulonephritis.

Original languageEnglish (US)
Pages (from-to)3901-3908
Number of pages8
JournalJournal of Immunology
Volume147
Issue number11
StatePublished - Dec 1 1991
Externally publishedYes

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CD55 Antigens
Mesangial Cells
Complement Activation
Antigen-Antibody Complex
Cell Membrane
Complement C5b
Complement C9
Serum
Staining and Labeling
Phosphoinositide Phospholipase C
Complement Membrane Attack Complex
Pronase

ASJC Scopus subject areas

  • Immunology

Cite this

Complement activation induces the expression of decay-accelerating factor on human mesangial cells. / Shibata, Toru; Cosio, Fernando G; Birmingham, Daniel J.

In: Journal of Immunology, Vol. 147, No. 11, 01.12.1991, p. 3901-3908.

Research output: Contribution to journalArticle

Shibata, T, Cosio, FG & Birmingham, DJ 1991, 'Complement activation induces the expression of decay-accelerating factor on human mesangial cells', Journal of Immunology, vol. 147, no. 11, pp. 3901-3908.
Shibata, Toru ; Cosio, Fernando G ; Birmingham, Daniel J. / Complement activation induces the expression of decay-accelerating factor on human mesangial cells. In: Journal of Immunology. 1991 ; Vol. 147, No. 11. pp. 3901-3908.
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abstract = "In the present study we evaluated the effect of complement activation by immune complexes (IC) on the expression of decay-accelerating factor (DAF) on human mesangial cells (MC). MC in culture were incubated with an Ag (DNP-Gelatin) that binds to fibronectin present in the MC matrix. Subsequently, MC were incubated with anti-DNP antibodies in the presence of human serum. By immunoperoxidase staining we showed that these incubations resulted in IC formation and deposition of human C3 and terminal complement components (C5b-9) on the mesangial matrix and on the surface of MC. By immunoperoxidase staining and by RIA we showed that IC formation and complement activation significantly increased DAF expression on the MC plasma membrane. The induction of DAF expression was a consequence of deposition of terminal complement components on the MC because, zymosan-activated serum and IC formation in the presence of C5- or C8-deficient serum failed to increase MC DAF expression. Furthermore, the observed increased DAF expression was the consequence of increased DAF synthesis by MC. Thus, both cycloheximide and actinomycin D blocked the increase on MC DAF observed after incubation with IC and serum. MC DAF had biophysical and functional characteristics similar to DAF in other cells. Thus, 1) MC DAF was resistant to trypsin but was removed from the MC membrane by pronase; 2) phosphatidylinositol-specific phospholipase C removed 48 ± 4{\%} of MC DAF indicating that MC DAF is anchored in the cell membrane by phosphatidylinositol groups; 3) DAF isolated from MC-inhibited complement-mediated hemolysis and demonstrated a molecular mass of 83 kDa. In conclusion, deposition of terminal complement components on human MC trigger new synthesis and membrane expression of DAF. Because DAF protects cells against complement-mediated lysis, we postulate that DAF may protect glomerular cells during IC and complement-mediated glomerulonephritis.",
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