Impairment of endothelial and subendothelial sites by a circulating plasma factor associated with minimal change disease

Abstract

The pathogenesis of albuminuria in minimal change disease (MCD) is unknown. A human plasma factor (denoted as 100KF) is able to induce minimal change-like glomerular alterations, i.e. loss of glomerular sialoglycoproteins and decreased expression of glomerular ecto—ATPase, following in vitro incubation with kidney tissue. In addition, increased (selective) permeability for plasma proteins occurs after perfusion of 100KF into the rat kidney ex vivo . As in kidney tissue from subjects with minimal change disease, subendothelial injury has also been observed, i.e. reduced anionic sites in the lamina rara interna, the question was raised whether injury induced by the plasma factor 100KF involves vascular endothelium or subendothelial matrix of the glomerular capillary wall. Permeability studies were carried out by using confluent endothelial monolayers (HUVEC) cultured on a standard two—compartment system. The permeability for a macromolecular marker (horse radish peroxidase) was tested following incubation of the monolayers with either the native plasma factor 100KF or the control factor (heat—inactivated 100KF), in combination with histochemical evaluation of the cells for ecto—ATPase expression. Also quantification of glomerular anionic sites at the ultrastructural level was carried out, after ex vivo perfusion of 100KF or control factor into rat kidneys. The plasma factor 100KF is able to increase the permeability of human endothelial monolayers for macromolecules in a dose—dependent manner (relative increase 122.4±24, 178.4±34 and 236.1 ±58% after preincubation with 0.05; 0.5 and 1.5 mg/ml 100KF respectively), concomitant with induction of minimal change-like histochemical alterations such as reduced expression of ecto—ATPase. The number of anionic sites in the lamina rara interna of the glomerular capillary wall is significantly diminished following perfusion with the plasma factor 100KF versus control factor (7.58±1.60 versus 12.57±2.05 per 1000nm; P <0.02); in contrast to the lamina rara externa (22.71±3.15 versus 22.27±2.92 per 1000 nm; statistically not significant). Endothelial cells and subendothelial matrix along the glomerular filtration barrier may be considered as target structures for the plasma factor 100KF, leading to initial minimal change-like alterations associated with glomerular albumin leakage.