title: Direct effect of plasma permeability factors from patients with idiopatic FSGS on nephrin and podocin expression in human podocytes.
abstract: The presence of circulating plasma factors (PF) altering renal permeability to proteins has been previously described in patients with focal segmental glomerulosclerosis (FSGS). Since these patients show reduced nephrin and podocin expression at renal biopsy, we evaluated the effect of serum and PF from patients with FSGS on nephrin and podocin expression in human podocytes. We studied 7 sera from patients with steroid-resistant FSGS, 3 from patients with nephrotic syndrome caused by non-immune disease, and 6 from healthy subjects. PF was prepared from plasmapheresis eluates of 2 patients with post-transplant recurrence of FSGS. Purification procedure was based on protein A Sepharose chromatography and differential precipitation in ammonium sulphate. Nephrin and podocin expression was semi-quantitatively evaluated by immunofluorescence. We found that serum and PF from FSGS patients rapidly induced redistribution and loss of nephrin in podocytes. This effect was associated with cytoskeleton redistribution and inhibited by cytochalasin B and sodium azide. On the contrary, podocin expression was unchanged after incubation with serum and PF from FSGS patients for short periods, but markedly reduced at 24 h. Our results demonstrate that serum and PF from FSGS patients may directly affect nephrin and podocin in human podocytes, thus providing new insights into the mechanisms causing proteinuria in FSGS.
authors.affiliation: Center of Experimental Research and Medical Sciences, Department of Internal Medicine, University of Turin, Turin, Italy.
authors.forename: Sophie
authors.initials: S
authors.lastname: Doublier
authors.forename: Luca
authors.initials: L
authors.lastname: Musante
authors.forename: Enrico
authors.initials: E
authors.lastname: Lupia
authors.forename: Giovanni
authors.initials: G
authors.lastname: Candiano
authors.forename: Tiziana
authors.initials: T
authors.lastname: Spatola
authors.forename: Gianluca
authors.initials: G
authors.lastname: Caridi
authors.forename: Cristina
authors.initials: C
authors.lastname: Zennaro
authors.forename: Michele
authors.initials: M
authors.lastname: Carraro
authors.forename: Gian Marco
authors.initials: GM
authors.lastname: Ghiggeri
authors.forename: Giovanni
authors.initials: G
authors.lastname: Camussi
mesh_terms: D000293:Adolescent; D000328:Adult; D001798:Blood Proteins / Q000378:metabolism*; D002478:Cells, Cultured; D002648:Child; D003599:Cytoskeleton / Q000378:metabolism / Q000473:pathology; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D015870:Gene Expression; D005923:Glomerulosclerosis, Focal Segmental / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism*; D008875:Middle Aged; D010539:Permeability; D050199:Podocytes / Q000378:metabolism*
chemical_list: D001798:Blood Proteins; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins; C407223:NPHS2 protein; C113267:nephrin