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Department of Nephrology, University Hospital Leiden, The
Netherlands
Department of Pathology, University Hospital Leiden, The
Netherlands
Division of Renal and Inflammatory Disease, Department of Medicine,
University Hospital, Nottingham, United Kingdom.
Correspondence to Dr. Mohamed R. Daha, Department of Nephrology, University Hospital Leiden, Building 1, C3-P, P. O. Box 9600, 2300 RC Leiden, The Netherlands. Phone: +31 71 5269364; Fax: +31 71 5248118; E-mail: M.R.Daha{at}Nephrology.MedFac.LeidenUniv.nl
Abstract. Mesangial cell (MC) injury is a characteristic feature in the early phase of Thy.1 nephritis. The present study investigates the contribution of complement to MC apoptosis in this experimental model of kidney disease in rats. Thy.1 nephritis was induced by injection of mouse anti-Thy.1 monoclonal antibody (ER4G). To assess the contribution of the terminal sequence of complement on apoptosis, the studies were performed in complement-sufficient PVG/c (PVG/c+) rats and in rats deficient in complement C6 (PVG/c-). Apoptosis was monitored by assessment of the number of condensed nuclei in kidney sections stained with periodic acid-Schiff (PAS) and by the terminal deoxynucleotidyl transferase-mediated nick end labeling (TUNEL) method and expressed as number of apoptotic cells per 50 glomerular cross sections. In the PAS method, 1 h after intravenous injection of ER4G, PVG/c+ rats exhibited 160.9 ± 49.5 apoptotic cells, whereas PVG/c- rats had only 3.2 ± 1.4 apoptotic cells. Control rats exhibited 0.9 ± 0.6 apoptotic cells. These findings were confirmed with the TUNEL method. In PVG/c- rats, a maximum number of 8.8 ± 3.1 TUNEL-positive (TUNEL+) cells was found at 6 h followed by a decline thereafter. In PVG/c+ rats, apoptosis was associated with deposition of C6 and C5b-9. Restoration of the complement system of PVG/c- rats with purified human C6 resulted in an increase of apoptosis at 1 h after injection of ER4G from minimal numbers to 239.9 ± 52.4 TUNEL+ cells. These studies appear to indicate for the first time that the terminal sequence of complement is involved in induction of apoptosis.
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