Autoantibodies to hLAMP-2 in ANCA-Negative Pauci-Immune Focal Necrotizing GN

Andrea Peschel, Neil Basu, Alexandra Benharkou, Ricarda Brandes, Markus Brown, Régis Dieckmann, Andrew J Rees, Renate Kain

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Abstract

Pauci-immune focal necrotizing GN (piFNGN) is usually associated with ANCAs that are thought to be pathogenic. However, 10%-15% of patients are ANCA negative and the cause of their injury is unknown. We previously reported a high frequency of autoantibodies to human lysosome-associated membrane protein-2 (hLAMP-2) in ANCA-associated piFNGN, and have now investigated whether the same is true in ANCA-negative patients. Of 11 patients, 8 (73%) had anti-hLAMP-2 antibodies detected by ELISA and confirmed by immunoblotting and indirect immunofluorescence. The autoantibodies from all 8 patients bound to native LAMP-2 purified from human glomeruli and recombinant hLAMP-2 expressed in ldlD cells, both with molecular masses of 110 kD. However, in contrast to anti-LAMP-2 antibodies from ANCA-positive patients, these antibodies from ANCA-negative patients failed to bind the more complexly glycosylated native neutrophil hLAMP-2 (190 kD). Treatment with the deglycosylating enzyme, endo-β-galactosidase, reduced the mass of neutrophil hLAMP-2 to 110 kD and enabled autoantibody binding. Similarly, pretreating neutrophils with endo-β-galactosidase or neuraminidase converted ANCA assay results from negative to positive. Finally, IgG from LAMP-2-positive ANCA-negative patients bound specifically to normal human kidney sections and to human glomerular endothelial cells in culture. In conclusion, in patients with ANCA-negative piFNGN, we have identified autoantibodies to hLAMP-2 that bind native glomerular but not neutrophil hLAMP-2, suggesting a role in pathogenesis.
Original languageEnglish
Pages (from-to)455-463
Number of pages9
JournalJournal of the American Society of Nephrology
Volume25
Issue number3
Early online date7 Nov 2013
DOIs
Publication statusPublished - Mar 2014

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Lysosome-Associated Membrane Glycoproteins
Antineutrophil Cytoplasmic Antibodies
Autoantibodies
Neutrophils
Galactosidases
Antibodies
Neuraminidase
Indirect Fluorescent Antibody Technique
Immunoblotting
Endothelial Cells
Cell Culture Techniques
Immunoglobulin G
Enzyme-Linked Immunosorbent Assay

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Peschel, A., Basu, N., Benharkou, A., Brandes, R., Brown, M., Dieckmann, R., ... Kain, R. (2014). Autoantibodies to hLAMP-2 in ANCA-Negative Pauci-Immune Focal Necrotizing GN. Journal of the American Society of Nephrology, 25(3), 455-463. https://doi.org/10.1681/ASN.2013030320

Autoantibodies to hLAMP-2 in ANCA-Negative Pauci-Immune Focal Necrotizing GN. / Peschel, Andrea; Basu, Neil; Benharkou, Alexandra; Brandes, Ricarda; Brown, Markus; Dieckmann, Régis; Rees, Andrew J; Kain, Renate.

In: Journal of the American Society of Nephrology, Vol. 25, No. 3, 03.2014, p. 455-463.

Research output: Contribution to journalArticle

Peschel, A, Basu, N, Benharkou, A, Brandes, R, Brown, M, Dieckmann, R, Rees, AJ & Kain, R 2014, 'Autoantibodies to hLAMP-2 in ANCA-Negative Pauci-Immune Focal Necrotizing GN' Journal of the American Society of Nephrology, vol. 25, no. 3, pp. 455-463. https://doi.org/10.1681/ASN.2013030320
Peschel, Andrea ; Basu, Neil ; Benharkou, Alexandra ; Brandes, Ricarda ; Brown, Markus ; Dieckmann, Régis ; Rees, Andrew J ; Kain, Renate. / Autoantibodies to hLAMP-2 in ANCA-Negative Pauci-Immune Focal Necrotizing GN. In: Journal of the American Society of Nephrology. 2014 ; Vol. 25, No. 3. pp. 455-463.
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