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Overexpression of Crry protects mesangial cells from complement-mediated injury.

M Nangaku, R J Quigg, S J Shankland, N Okada, R J Johnson and W G Couser
JASN February 1997, 8 (2) 223-233; DOI: https://doi.org/10.1681/ASN.V82223
M Nangaku
Division of Nephrology, University of Washington, Seattle 98195-6521, USA.
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R J Quigg
Division of Nephrology, University of Washington, Seattle 98195-6521, USA.
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S J Shankland
Division of Nephrology, University of Washington, Seattle 98195-6521, USA.
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N Okada
Division of Nephrology, University of Washington, Seattle 98195-6521, USA.
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R J Johnson
Division of Nephrology, University of Washington, Seattle 98195-6521, USA.
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W G Couser
Division of Nephrology, University of Washington, Seattle 98195-6521, USA.
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Abstract

Crry is a membrane-associated complement regulatory protein expressed on glomerular mesangial, endothelial, and epithelial cells, which reduces C3/C5 convertase activity. This study utilized an overexpression strategy to determine the functional significance of Crry in cultured rat mesangial cells. A Crry expression vector was constructed and was tagged with a c-myc epitope that allowed transfected Crry to be distinguished from the constitutively expressed protein. In stable clones, overexpressed Crry was clearly detected immunocytochemically both by anti-c-myc and anti-Crry antibody in a membrane localization. The overexpression of Crry was also confirmed by Western blotting and immunoprecipitation. To determine if overexpression of Crry by mesangial cells confers a protective effect from complement attack, complement-mediated cell lysis assays were performed. Crry-transfected mesangial cells demonstrated complete resistance to complement-mediated cell lysis, which was reversed by neutralization of Crry with both monoclonal antibody and F(ab')2 fragments of the antibody. This study also investigated the role of Crry in protecting cells from the effects of sublytic complement attack. Overexpressed Crry suppressed antibody/complement induced production of superoxide, one of the inflammatory mediators induced by sublytic complement attack. Immunocytochemical staining confirmed a reduction in C3 and C5b-9 deposition in Crry-transfected cells. These results demonstrate directly that transfected Crry functions as a potent protector of mesangial cells against complement-mediated injury. Crry may play an important role in modulating the glomerular response to immune injury in vivo.

  • Copyright © 1997 by American Society of Nephrology
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Journal of the American Society of Nephrology
Vol. 8, Issue 2
1 Feb 1997
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Overexpression of Crry protects mesangial cells from complement-mediated injury.
M Nangaku, R J Quigg, S J Shankland, N Okada, R J Johnson, W G Couser
JASN Feb 1997, 8 (2) 223-233; DOI: 10.1681/ASN.V82223

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Overexpression of Crry protects mesangial cells from complement-mediated injury.
M Nangaku, R J Quigg, S J Shankland, N Okada, R J Johnson, W G Couser
JASN Feb 1997, 8 (2) 223-233; DOI: 10.1681/ASN.V82223
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