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Routine immunohistochemical staining in membranous nephropathy: in situ detection of phospholipase A2 receptor and thrombospondin type 1 containing 7A domain

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Abstract

Background

Membranous nephropathy (MN) can be idiopathic (iMN) or manifest as a result of systemic underlying conditions as a secondary epiphenomenon. For the prognostic and predictive consequences of this discrimination, the routine use of reliable markers is crucial. This large MN series aimed to evaluate the routine and standardized immunohistochemical (IHC) employment of a panel of 3 biomarkers—phospholipase A2 receptor (PLA2R), thrombospondin type-1 domain-containing 7A (THSD7A), and immunoglobulin (Ig)G4—in the differential diagnosis of MN forms, contributing to the validation of the technique and the correct interpretation of reproducible patterns of reactivity.

Methods

We classified 95 patients with a biopsy proven diagnosis of MN as primary (n = 72) or secondary (n = 23) cases based on clinical data. After performing an IHC assay directed against PLA2R, THSD7A and IgG4 antigens, samples were interpreted by three different nephropathologists to assess the positivity/negativity of the staining according to new interpretation criteria.

Results

Useful interpretation criteria were introduced to exclude false positive patterns of reactivity and to identify only true granular membranous or mesangial deposits in MN. The IHC directed against PLA2R resulted positive in 51 iMN cases and negative in 21, while 4/23 secondary forms were considered positive. Based on these data the technique showed a sensitivity of 71% and specificity of 83%. On the other hand, the IHC analysis for IgG4 resulted positive in 44 cases of iMN and negative in 28 cases, while only 4/23 secondary forms were positive (same cases positive to PLA2R). Finally, THSD7A was found to be positive only in 1 case, which was negative to PLA2R and IgG4. The combination of the results allowed a classification of the series into two major groups: “double-positive” (PLA2R+/IgG4+/THSD7A−) and “triple-negative” (PLA2R−/IgG4−/THSD7A−) cases.

Conclusions

Based on these data, the diagnostic performance of the three biomarkers used in a “tandem fashion” can reach 79% sensitivity and 83% specificity, significantly reducing the risk of a false-positive or false-negative result and improving the routine characterization of this frequent glomerulonephritis.

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Acknowledgements

Our special thanks go to Carla Scalia, Loredana Tusa and Lorella Riva for their technical support and to nephrological team of the San Gerardo Hospital for the routine execution of the renal biopsies.

Funding

No support was used for this study.

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Authors and Affiliations

  1. Department of Medicine and Surgery, Pathology, University of Milano-Bicocca, San Gerardo Hospital, Monza, Italy

    Vincenzo L’Imperio & Fabio Pagni

  2. Nephrology Unit, Department of Medicine and Surgery, University of Milano-Bicocca, Monza, Italy

    Federico Pieruzzi & Renato Alberto Sinico

  3. Department of Biomedical and Clinical Sciences L. Sacco, University of Milan, Milan, Italy

    Manuela Nebuloni

  4. Research Center for Renal Immunopathology, University of Milan, Milan, Italy

    Manuela Nebuloni & Fabio Pagni

  5. Department of Nephrology, San Giovanni Di Dio Hospital, Agrigento, Italy

    Antonio Granata

  6. Proteomics and Metabolomics Unit, Department of Medicine and Surgery, University of Milano-Bicocca, Monza, Italy

    Andrew Smith

  7. Microbiology and Virology Department, San Carlo Borromeo Hospital, Milan, Italy

    Antonella Radice

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  1. Vincenzo L’Imperio
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  2. Federico Pieruzzi
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Correspondence to Fabio Pagni.

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All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

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L’Imperio, V., Pieruzzi, F., Sinico, R.A. et al. Routine immunohistochemical staining in membranous nephropathy: in situ detection of phospholipase A2 receptor and thrombospondin type 1 containing 7A domain. J Nephrol 31, 543–550 (2018). https://doi.org/10.1007/s40620-018-0489-z

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