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. 2014 Jul;6(2):76-9.
doi: 10.4103/0974-2727.141500.

C4d immunohistochemistry in membranous nephropathy

Monalisa Hui  1 , Megha S Uppin  1 , Aruna K Prayaga  1 , Sree Bhushan Raju  2 , Liza Rajasekhar  3
Affiliations

C4d immunohistochemistry in membranous nephropathy

Monalisa Hui et al. J Lab Physicians. 2014 Jul.

Abstract

Background: Membranous nephropathy (MN) is the most common cause of nephropathy in adults. The diagnosis is based on characteristic light microscopic, electron microscope and immunofluorescence (IF) findings. In early MN, the light microscopic findings may be difficult to differentiate from minimal chain disease. In the absence of fresh frozen tissue for IF, immunohistochemistry with C4d aids in the diagnosis.

Materials and methods: A total 48 cases of MN diagnosed on renal biopsy were analyzed. The formalin fixed paraffin embedded tissues were stained with routine hematoxylin and eosin stains along with periodic acid-Schiff and silver methenamine stains to highlight the basement membrane. Fresh frozen tissues were available for IF in 40 cases. Immunostaining with C4d was done on paraffin-embedded sections by polymer-Horse Radish Peroxidase (HRP) technique using polyclonal antiserum to C4d (Biogenex, India).

Results: There were 25 cases of idiopathic MN, 17 cases of Class V lupus nephritis and 2 cases were secondary to hepatitis C infection with cirrhosis. The glomerular basement membrane (GBM) was diffusely thickened with formation of spikes in 28 cases. In 11 cases the capillary loops were rigid but spikes were not seen and in 9 cases there was no apparent thickening of the basement membrane. All the cases showed diffuse positivity for C4d along the GBM.

Conclusion: C4d is a reliable method to establish the diagnosis of MN and also a sensitive marker of complement activation reflecting the pathogenesis of MN.

Keywords: C4d; complement activation; immunohistochemistry; membranous nephropathy.

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Conflict of interest statement

Conflict of Interest: None declared.

Figures

Figure 1
Figure 1
Classic case of membranous nephropathy with (a) thickened basement membrane, (b) showing spikes on silver methanamine-periodic acid-Schiff (SM-PAS) stain and (c) granular positivity for C4d along the basement membrane. Horseradish peroxidase polymer; C4d. Another biopsy of a 36/M with 24 h proteinuria of 3.5 g. The glomerulus on SM-PAS stain, (d and e) showing almost normal glomerulus without apparent thickening of the basement membrane, (f) immunofluorescence showed granular positivity for immunoglobulin G, (g) however C4d showed positivity along the glomerular basement membrane
Figure 2
Figure 2
(a) A case of class IV lupus nephritis with diffuse proliferation and neutrophils, (b) C4d positivity along the glomerular basement membrane
See this image and copyright information in PMC

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