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Characterization of THSD7A-antibodies not binding to glomerular THSD7A in a patient with diabetes mellitus but no membranous nephropathy

Membranous nephropathy (MN) is an autoimmune disease caused by autoantibodies against the podocyte antigens phospholipase A(2) receptor 1 (PLA(2)R1) and thrombospondin type 1 domain containing protein 7A (THSD7A) in 80% and 2–3% of patients, respectively. THSD7A antibodies are considered to be patho...

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Autores principales: Reinhard, Linda, Thomas, Cindy, Machalitza, Maya, Lattwein, Erik, Weiss, Lothar S., Vitu, Jan, Wiech, Thorsten, Stahl, Rolf A. K., Hoxha, Elion
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8355258/
https://www.ncbi.nlm.nih.gov/pubmed/34376704
http://dx.doi.org/10.1038/s41598-021-94921-y
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author Reinhard, Linda
Thomas, Cindy
Machalitza, Maya
Lattwein, Erik
Weiss, Lothar S.
Vitu, Jan
Wiech, Thorsten
Stahl, Rolf A. K.
Hoxha, Elion
author_facet Reinhard, Linda
Thomas, Cindy
Machalitza, Maya
Lattwein, Erik
Weiss, Lothar S.
Vitu, Jan
Wiech, Thorsten
Stahl, Rolf A. K.
Hoxha, Elion
author_sort Reinhard, Linda
collection PubMed
description Membranous nephropathy (MN) is an autoimmune disease caused by autoantibodies against the podocyte antigens phospholipase A(2) receptor 1 (PLA(2)R1) and thrombospondin type 1 domain containing protein 7A (THSD7A) in 80% and 2–3% of patients, respectively. THSD7A antibodies are considered to be pathogenic and highly specific for MN patients. Using an indirect immunofluorescence test (IIFT) we detected THSD7A-antibodies (titre 1:10) in the serum of a patient with high proteinuria who, however, in the kidney biopsy was diagnosed with diabetic nephropathy and MN was excluded as a possible cause of proteinuria. Different immunofluorescence assays and Western blot techniques using recombinant THSD7A (rTHSD7A) or THSD7A from different human tissues revealed that the circulating THSD7A-autoantibodies were only of the IgG3 subclass. The patient serum reacted exclusively with rTHSD7A and only when the antigen was present in reducing Western blot conditions, or on formaldehyde-fixed cells for the IIFT. Our findings show for the first time the existence of circulating THSD7A-antibodies recognizing denatured/reduced rTHSD7A, which do not react with glomerular THSD7A in vivo and are thus presumptively non-pathogenic. As a consequence, kidney biopsy or Western blot analyses of THSD7A under non-reducing conditions should be performed to confirm the diagnosis of THSD7A-associated MN, especially in cases with low THSD7A-antibody levels in the IIFT.
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spelling pubmed-83552582021-08-11 Characterization of THSD7A-antibodies not binding to glomerular THSD7A in a patient with diabetes mellitus but no membranous nephropathy Reinhard, Linda Thomas, Cindy Machalitza, Maya Lattwein, Erik Weiss, Lothar S. Vitu, Jan Wiech, Thorsten Stahl, Rolf A. K. Hoxha, Elion Sci Rep Article Membranous nephropathy (MN) is an autoimmune disease caused by autoantibodies against the podocyte antigens phospholipase A(2) receptor 1 (PLA(2)R1) and thrombospondin type 1 domain containing protein 7A (THSD7A) in 80% and 2–3% of patients, respectively. THSD7A antibodies are considered to be pathogenic and highly specific for MN patients. Using an indirect immunofluorescence test (IIFT) we detected THSD7A-antibodies (titre 1:10) in the serum of a patient with high proteinuria who, however, in the kidney biopsy was diagnosed with diabetic nephropathy and MN was excluded as a possible cause of proteinuria. Different immunofluorescence assays and Western blot techniques using recombinant THSD7A (rTHSD7A) or THSD7A from different human tissues revealed that the circulating THSD7A-autoantibodies were only of the IgG3 subclass. The patient serum reacted exclusively with rTHSD7A and only when the antigen was present in reducing Western blot conditions, or on formaldehyde-fixed cells for the IIFT. Our findings show for the first time the existence of circulating THSD7A-antibodies recognizing denatured/reduced rTHSD7A, which do not react with glomerular THSD7A in vivo and are thus presumptively non-pathogenic. As a consequence, kidney biopsy or Western blot analyses of THSD7A under non-reducing conditions should be performed to confirm the diagnosis of THSD7A-associated MN, especially in cases with low THSD7A-antibody levels in the IIFT. Nature Publishing Group UK 2021-08-10 /pmc/articles/PMC8355258/ /pubmed/34376704 http://dx.doi.org/10.1038/s41598-021-94921-y Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Reinhard, Linda
Thomas, Cindy
Machalitza, Maya
Lattwein, Erik
Weiss, Lothar S.
Vitu, Jan
Wiech, Thorsten
Stahl, Rolf A. K.
Hoxha, Elion
Characterization of THSD7A-antibodies not binding to glomerular THSD7A in a patient with diabetes mellitus but no membranous nephropathy
title Characterization of THSD7A-antibodies not binding to glomerular THSD7A in a patient with diabetes mellitus but no membranous nephropathy
title_full Characterization of THSD7A-antibodies not binding to glomerular THSD7A in a patient with diabetes mellitus but no membranous nephropathy
title_fullStr Characterization of THSD7A-antibodies not binding to glomerular THSD7A in a patient with diabetes mellitus but no membranous nephropathy
title_full_unstemmed Characterization of THSD7A-antibodies not binding to glomerular THSD7A in a patient with diabetes mellitus but no membranous nephropathy
title_short Characterization of THSD7A-antibodies not binding to glomerular THSD7A in a patient with diabetes mellitus but no membranous nephropathy
title_sort characterization of thsd7a-antibodies not binding to glomerular thsd7a in a patient with diabetes mellitus but no membranous nephropathy
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8355258/
https://www.ncbi.nlm.nih.gov/pubmed/34376704
http://dx.doi.org/10.1038/s41598-021-94921-y
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