Pathogenic autoantibodies from patients with lupus nephritis cause reduced tyrosine phosphorylation of podocyte proteins, including tubulin.
BACKGROUND
The tertiary structure of normal podocytes prevents protein from leaking into urine. Patients with lupus nephritis (LN) develop proteinuria, and kidney biopsies from these patients display a number of podocyte abnormalities including retraction of podocyte processes. Autoantibodies have been shown to deposit in the kidneys of patients and mice with LN and are believed to play a key role in causing renal inflammation and dysfunction. The objective of this research was to study the effects of IgG antibodies from patients with LN on cultured human podocytes.


METHODS
We exposed a human podocyte cell line to heat-inactivated (HI) plasma and purified polyclonal IgG from the following groups of subjects; patients with LN, patients with lupus without nephritis, patients with rheumatoid arthritis and healthy controls. We measured expression and intracellular distribution of podocyte-specific proteins and global phosphorylation of tyrosine. We then used mass spectrometry to identify the major protein targets of this phosphorylation.


RESULTS
HI LN plasma did not alter expression or cellular distribution of podocyte-specific proteins but caused a significant reduction in podocyte protein tyrosine phosphorylation compared with plasma from healthy controls (p=0.0008). This result was replicated using purified IgG but was not seen with plasma from rheumatoid arthritis or non-renal lupus patients. The dominant tyrosine phosphorylated protein in podocytes was 55 kDa in size and was identified as tubulin.


CONCLUSIONS
Since tubulin is an important component of podocyte major processes, these results suggest that autoantibodies from LN patients may exert an important pathogenic effect by dephosphorylation of this protein in podocytes.
Division of Medicine , Centre for Rheumatology Research, University College London, Rayne Institute , London , UK.
Jessica J
JJ
Manson
Clinical and Molecular Genetics Unit , Institute for Child Health and Great Ormond Street Hospital , London , UK.
Kevin
K
Mills
Division of Medicine , Centre for Rheumatology Research, University College London, Rayne Institute , London , UK.
Elizabeth
E
Jury
Division of Medicine , Centre for Rheumatology Research, University College London, Rayne Institute , London , UK.
Lesley
L
Mason
Lupus Research Unit , St Thomas' Hospital , London , UK.
David P
DP
D'Cruz
Department of Renal Medicine , University of Bristol , Bristol , UK.
Lan
L
Ni
Department of Renal Medicine , University of Bristol , Bristol , UK.
Moin
M
Saleem
Department of Renal Medicine , University of Bristol , Bristol , UK.
Peter
P
Mathieson
Division of Medicine , Centre for Rheumatology Research, University College London, Rayne Institute , London , UK.
David
D
Isenberg
Division of Medicine , Centre for Rheumatology Research, University College London, Rayne Institute , London , UK.
Anisur
A
Rahman
nephritis; phosphorylation; podocyte