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American Journal of Pathology, Vol 142, 529-539, Copyright © 1993 by American Society for Investigative Pathology

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Rat mesangial cell lysis in vitro is induced by cationic polypeptides

JA Broestl and SN Emancipator
Department of Pathology, Case Western Reserve University School of Medicine, Cleveland, Ohio.

Immunization and challenge with cationic proteins induces immune complex glomerulonephritis in rodents. Cationic (c) bovine gamma- globulin (BGG), when added to rat mesangial cells in vitro, induced release of lysosomal beta-glucuronidase, an enzyme capable of degrading basement membrane components. Native (n) BGG, alone or in the presence of specific antibody, did not. Morphological changes (cellular swelling) in response to cBGG suggested cell injury; indeed, significant lactate dehydrogenase (LDH) was released into the media, depending on dose, time, calcium, and temperature. Prior trypsinization of cBGG abrogated this response. The synthetic polycation, poly L- lysine > 50 kd, similarly elicited LDH release; 4-kd poly L-lysine or protamine sulfate had little or no effect. Anionic heparin sodium inhibited cBGG-induced morphological changes and, when coincubated with cBGG, significantly reduced LDH release (P < 0.0001) to levels equal to or less than those with the nBGG control. This heparin effect was lost if addition was delayed until 10 minutes after the addition of cBGG, indicating an irreversible effect of cBGG within this time. We conclude that charge alone is not sufficient for polycations to induce LDH release. Moreover, the cellular swelling and rapidity of LDH release suggest that cytotoxicity results from direct plasma membrane destruction, perhaps due to altered sodium ion concentrations.




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Copyright © 1993 by the American Society for Investigative Pathology.