Aquaporin-4 autoantibodies in neuromyelitis optica

AQP4 isoform-dependent sensitivity and specificity

Francesco Pisani, Angelo Sparaneo, Carla Tortorella, Maddalena Ruggieri, Maria Trojano, Maria Grazia Mola, Grazia Paola Nicchia, Antonio Frigeri, Maria Svelto

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Neuromyelitis Optica (NMO) is an autoimmune demyelinating disease, characterized by the presence of autoantibody (NMO-IgG) to Aquaporin-4 (AQP4). NMO-IgG identification supports NMO diagnosis and several diagnostic tests have been developed, but their sensitivity is too variable, and some assay show low sensitivity. This impairs correct diagnosis of NMO. By cell based assay (CBA) we here evaluate the efficacy of different strategies to express AQP4 in mammalian cells in terms of: a) AQP4 translation initiation signals; b) AQP4 isoforms (M1 and M23) and fluorescent tag position; c) NMO serum concentration and AQP4 degradation. Our results demonstrate that when using AQP4-M1, the nucleotide in position -3 of the AUG greatly affects the AQP4-M1/M23 protein ratio, NMO-IgG binding, and consequently test sensitivity. Test sensitivity was highest with M23 expressing cells (97.5%) and only 27.5% with AQP4-M1. The fluorescent tag added to the N-terminus of AQP4-M23 considerably affected the NMO-IgG binding, and test sensitivity, due to disruption of AQP4 suprastructures. Furthermore, sera used at high concentration resulted in AQP4 degradation which affected test sensitivity. To further evaluate the reliability of the M23 based CBA test, samples of one NMO patient collected during about 2 years clinical follow-up were tested. The results of serum titer correlated with disease activity and treatment response. In conclusion, we provide a molecular explanation for the contrasting CBA test data reported and suggest the use of M23 with a C-terminus fluorescent tag as the proper test for NMO diagnosis.

Original languageEnglish
Article numbere79185
JournalPLoS One
Volume8
Issue number11
DOIs
Publication statusPublished - Nov 15 2013

Fingerprint

Aquaporin 4
Neuromyelitis Optica
autoantibodies
aquaporins
Autoantibodies
Protein Isoforms
Sensitivity and Specificity
Assays
Immunoglobulin G
testing
assays
cells
Serum
Degradation
degradation
autoimmune diseases
Demyelinating Diseases
Routine Diagnostic Tests
diagnostic techniques
Autoimmune Diseases

ASJC Scopus subject areas

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

Cite this

Pisani, F., Sparaneo, A., Tortorella, C., Ruggieri, M., Trojano, M., Mola, M. G., ... Svelto, M. (2013). Aquaporin-4 autoantibodies in neuromyelitis optica: AQP4 isoform-dependent sensitivity and specificity. PLoS One, 8(11), [e79185]. https://doi.org/10.1371/journal.pone.0079185

Aquaporin-4 autoantibodies in neuromyelitis optica : AQP4 isoform-dependent sensitivity and specificity. / Pisani, Francesco; Sparaneo, Angelo; Tortorella, Carla; Ruggieri, Maddalena; Trojano, Maria; Mola, Maria Grazia; Nicchia, Grazia Paola; Frigeri, Antonio; Svelto, Maria.

In: PLoS One, Vol. 8, No. 11, e79185, 15.11.2013.

Research output: Contribution to journalArticle

Pisani, F, Sparaneo, A, Tortorella, C, Ruggieri, M, Trojano, M, Mola, MG, Nicchia, GP, Frigeri, A & Svelto, M 2013, 'Aquaporin-4 autoantibodies in neuromyelitis optica: AQP4 isoform-dependent sensitivity and specificity', PLoS One, vol. 8, no. 11, e79185. https://doi.org/10.1371/journal.pone.0079185
Pisani, Francesco ; Sparaneo, Angelo ; Tortorella, Carla ; Ruggieri, Maddalena ; Trojano, Maria ; Mola, Maria Grazia ; Nicchia, Grazia Paola ; Frigeri, Antonio ; Svelto, Maria. / Aquaporin-4 autoantibodies in neuromyelitis optica : AQP4 isoform-dependent sensitivity and specificity. In: PLoS One. 2013 ; Vol. 8, No. 11.
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