Identification and Characterization of a New Autoimmune Protein in Membranous Nephropathy by Immunoscreening of a Renal cDNA Library

Fabrizio Cavazzini, Riccardo Magistroni, Luciana Furci, Valentina Lupo, Giulia Ligabue, Maria Granito, Marco Leonelli, Alberto Albertazzi, Gianni Cappelli

Research output: Contribution to journalArticle

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.

Original languageEnglish
Article numbere48845
JournalPLoS One
Volume7
Issue number11
DOIs
Publication statusPublished - Nov 8 2012

Fingerprint

Membranous Glomerulonephritis
kidney diseases
Gene Library
cDNA libraries
kidneys
Kidney
Biopsy
Synaptonemal Complex
synaptonemal complex
biopsy
Proteins
proteins
Serum
nephrotic syndrome
antigens
Bacteriophage lambda
Antigens
Control Groups
Bacteriophages
antibodies

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Cavazzini, F., Magistroni, R., Furci, L., Lupo, V., Ligabue, G., Granito, M., ... Cappelli, G. (2012). Identification and Characterization of a New Autoimmune Protein in Membranous Nephropathy by Immunoscreening of a Renal cDNA Library. PLoS One, 7(11), [e48845]. https://doi.org/10.1371/journal.pone.0048845

Identification and Characterization of a New Autoimmune Protein in Membranous Nephropathy by Immunoscreening of a Renal cDNA Library. / Cavazzini, Fabrizio; Magistroni, Riccardo; Furci, Luciana; Lupo, Valentina; Ligabue, Giulia; Granito, Maria; Leonelli, Marco; Albertazzi, Alberto; Cappelli, Gianni.

In: PLoS One, Vol. 7, No. 11, e48845, 08.11.2012.

Research output: Contribution to journalArticle

Cavazzini, F, Magistroni, R, Furci, L, Lupo, V, Ligabue, G, Granito, M, Leonelli, M, Albertazzi, A & Cappelli, G 2012, 'Identification and Characterization of a New Autoimmune Protein in Membranous Nephropathy by Immunoscreening of a Renal cDNA Library', PLoS One, vol. 7, no. 11, e48845. https://doi.org/10.1371/journal.pone.0048845
Cavazzini, Fabrizio ; Magistroni, Riccardo ; Furci, Luciana ; Lupo, Valentina ; Ligabue, Giulia ; Granito, Maria ; Leonelli, Marco ; Albertazzi, Alberto ; Cappelli, Gianni. / Identification and Characterization of a New Autoimmune Protein in Membranous Nephropathy by Immunoscreening of a Renal cDNA Library. In: PLoS One. 2012 ; Vol. 7, No. 11.
@article{a43774d2ec3a44c3ba8c82de0d0b0745,
title = "Identification and Characterization of a New Autoimmune Protein in Membranous Nephropathy by Immunoscreening of a Renal cDNA Library",
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.",
author = "Fabrizio Cavazzini and Riccardo Magistroni and Luciana Furci and Valentina Lupo and Giulia Ligabue and Maria Granito and Marco Leonelli and Alberto Albertazzi and Gianni Cappelli",
year = "2012",
month = "11",
day = "8",
doi = "10.1371/journal.pone.0048845",
language = "English",
volume = "7",
journal = "PLoS One",
issn = "1932-6203",
publisher = "Public Library of Science",
number = "11",

}

TY - JOUR

T1 - Identification and Characterization of a New Autoimmune Protein in Membranous Nephropathy by Immunoscreening of a Renal cDNA Library

AU - Cavazzini, Fabrizio

AU - Magistroni, Riccardo

AU - Furci, Luciana

AU - Lupo, Valentina

AU - Ligabue, Giulia

AU - Granito, Maria

AU - Leonelli, Marco

AU - Albertazzi, Alberto

AU - Cappelli, Gianni

PY - 2012/11/8

Y1 - 2012/11/8

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=84869004122&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84869004122&partnerID=8YFLogxK

U2 - 10.1371/journal.pone.0048845

DO - 10.1371/journal.pone.0048845

M3 - Article

C2 - 23144993

AN - SCOPUS:84869004122

VL - 7

JO - PLoS One

JF - PLoS One

SN - 1932-6203

IS - 11

M1 - e48845

ER -