Punctate IgG staining particles localize in the budding ballooning clusters of reactive foot processes in minimal change disease
Ping L. ZhangVasudevan D. MahalingamBrandon D. MetcalfYazan Al-OthmanWei LiHassan D. KanaanGuillermo A. Herrera
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Abstract:
The etiology of minimal change disease (MCD) remains a mystery as the only characteristic findings are the diffuse effacement of foot processes seen on electron microscopy (EM). Punctate IgG staining found floating outside glomerular capillary loops in MCD cases was recently identified as autoimmune antibodies against nephrin of podocytes. We hypothesized that the punctate IgG staining is located on budding ballooning clusters (BBC) of reactive foot processes in Bowman’s space found on EM. We identified seven patients with MCD cases showing IgG staining that were subsequently evaluated for BBC on EM. We concurrently examined 12 negative controls, either unremarkable cases or tubulointerstitial diseases, by EM. Immunogold labeling was performed to confirm the presence of IgG and determine localization. In seven MCD cases, there were positive punctate IgG staining particles outside of the glomerular basement membranes (GBM) along with concurrent punctate staining for C3, kappa, and lambda. By EM, all seven (100%) MCD cases revealed BBC that was characterized by ballooning foot processes ranging from 1 to 6 µm and was either budding or detached from the GBM in 3–7 clusters; no electron-dense materials were seen in BBC. BBC was also seen in only 1 of 12 (8%) negative controls. Immunogold labeling identified IgG particles within BBC of MCD by EM, but not in the negative control. Our data suggest that BBC are EM structures of reactive foot processes that are most likely correlated with punctate IgG staining seen in cases of MCD, supported by immunogold labeling for IgG.Keywords:
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Electron microscopy permits the detailed study of cell relationships within tissues and organelles within cells. Electron immunocytochemistry is the high resolution study of antigens within cells and their relation to cell ultrastructure. Fixation to achieve optimal fine structural detail for electron microscopy is exactly that which damages antigens with respect to reaction with specific antibody. Cell preparation for electron microscopy is therefore a compromise between retaining sufficient antigenicity while preserving the cell ultrastructure.
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Effects of Lugol's iodine pretreatment upon immunogold-silver staining were examined in terms of the intensification of the reaction products obtained by physical development. Iodine pretreatment has diminished the duration necessary for the development, whereas the staining reaction of an appropriate intensity could hardly be obtained by the pretreatment. In addition, essential staining reactions tended to be accompanied by marked nonspecific silver precipitations. It is concluded that iodine pretreatment should either be omitted or used with cautions in immunogold-silver staining procedures for the precise detection of antigenic sites.
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Journal Article Immunogold Labeling of Cultured Cells and Virus Particles for Electron Microscopy and Cryo-Electron Microscopy Applications. Get access Hong Yi, Hong Yi Robert P. Apkarian Integrated Electron Microscopy Core, Emory University, Atlanta, Georgia, USA Search for other works by this author on: Oxford Academic Google Scholar Joshua D Strauss, Joshua D Strauss Department of Pediatrics, Emory University School of Medicine, Children’s Healthcare of Atlanta,Atlanta, Georgia, USA Search for other works by this author on: Oxford Academic Google Scholar Jason E Hammonds, Jason E Hammonds Department of Pediatrics, Emory University School of Medicine, Children’s Healthcare of Atlanta,Atlanta, Georgia, USA Search for other works by this author on: Oxford Academic Google Scholar Ravi Dyavar Shetty, Ravi Dyavar Shetty Department of Microbiology and Immunology, Emory University, Atlanta, Georgia, USA Search for other works by this author on: Oxford Academic Google Scholar Rama R Amara, Rama R Amara Department of Microbiology and Immunology, Emory University, Atlanta, Georgia, USA Search for other works by this author on: Oxford Academic Google Scholar Paul W Spearman, Paul W Spearman Department of Pediatrics, Emory University School of Medicine, Children’s Healthcare of Atlanta,Atlanta, Georgia, USA Search for other works by this author on: Oxford Academic Google Scholar Elizabeth R Wright Elizabeth R Wright Robert P. Apkarian Integrated Electron Microscopy Core, Emory University, Atlanta, Georgia, USADepartment of Pediatrics, Emory University School of Medicine, Children’s Healthcare of Atlanta,Atlanta, Georgia, USA Search for other works by this author on: Oxford Academic Google Scholar Microscopy and Microanalysis, Volume 20, Issue S3, 1 August 2014, Pages 1220–1221, https://doi.org/10.1017/S1431927614007831 Published: 27 August 2014
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