Identification and characterization of a new autoimmune protein in membranous nephropathy by immunoscreening of a renal cDNA library

PLoS One. 2012;7(11):e48845. doi: 10.1371/journal.pone.0048845. Epub 2012 Nov 8.

Abstract

Membranous Nephropathy (MN) represents a large amount of Nephrotic Syndromes in the adult population and its definitive diagnosis is currently carried out through biopsy. An autoimmune condition has been demonstrated in idiopathic MN (iMN) in which some kidney structures are targeted by patient autoantibodies. Some candidate antigens have been described and other likely involved target proteins responsible for the disease are not known yet. In this work our aim is to identify these proteins by screening a lambda-phage library with patients' sera. We enrolled four groups of patients: two MN groups of 12 full iMN patients; one control group of 15 patients suffering from other renal diseases; one control group of 15 healthy individuals. A commercial cDNA phagemide library was screened using the above described sera, in order to detect positive signals due to antigen-antibody bond. We detected one phagemide clone expressing a protein which was shown to be targeted by the antibodies of the iMN sera only. Control sera were negative. The sequence analysis of cDNA matched the Synaptonemal Complex protein 65 (SC65) coding sequence. Further proteomic analyses were carried out to validate our results. We provide evidence of an involvement of SC65 protein as an autoimmune target in iMN. Considering the invasiveness and the resulting risk coming from renal biopsy, our ongoing aim is to set a procedure able to diagnose affected patients through a little- or non-invasive method such as blood sampling rather than biopsy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Aged, 80 and over
  • Autoantibodies / immunology*
  • Autoantibodies / metabolism
  • Autoantibodies / physiology
  • Autoantigens / chemistry
  • Autoantigens / genetics
  • Autoantigens / immunology
  • Autoantigens / metabolism*
  • Autoantigens / physiology
  • Biopsy
  • Blotting, Western
  • DNA, Complementary / chemistry
  • Female
  • Fluorescent Antibody Technique
  • Gene Library
  • Glomerulonephritis, Membranous / immunology*
  • Humans
  • Kidney / pathology
  • Male
  • Middle Aged
  • Polymorphism, Restriction Fragment Length
  • Sequence Analysis, DNA
  • Synaptonemal Complex

Substances

  • Autoantibodies
  • Autoantigens
  • DNA, Complementary
  • P3H4 protein, human

Grants and funding

This work was supported by grant 2004064212 from the Italian Ministry of Education, University and Research (MIUR). The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.