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. Author manuscript; available in PMC: 2015 Mar 25.
Published in final edited form as: J Immunol. 2014 Oct 22;193(11):5434–5443. doi: 10.4049/jimmunol.1401462

Figure 3. IgA deposition in kidneys of C4-/- mice is positively correlated with dsDNA-specific IgA serum levels.

Figure 3

A) Representative glomerular IgA staining in kidneys harvested from wild type and C4-/- mice 5 months post-Pneumovax (Px) immunization alone (left panels) or 5 months post-Pneumovax (Px) immunization with i.p. WU2 infection challenge on d14 (middle panels). IgA staining of aged (9-10 month-old) naïve wild type and C4-/-mice is shown (right panels). FITC-conjugated goat anti-mouse IgA was used for detection (400X magnification). B) Quantitation of mean IgA and Ig fluorescence (mean fluorescence intensity) in kidney sections of C4+/+ and C4-/- mice 5 months post Pneumovax immunization (Px), 5 months post Pneumovax immunization with i.p. WU2 infection on d14 (Px + infx), and age-matched naïve C4-/- mice. Fluorescence intensity of glomerular Ig staining was quantified using ImageJ software and expressed as density per unit area (with the average fluorescence measured from 10 or more glomeruli). Values for individual mice are shown. Mean fluorescent intensity is indicated as horizontal bars. Significant differences in means are indicated (*p<0.05). C) Graphs depict the association (pairwise comparison) between the mean fluorescence intensity signals of IgA staining in kidneys (as shown in A and quantitated in B) and the levels of dsDNA-specific serum IgA 5 months post immunization measured as absorbance values obtained at a 1:100 dilution (left panel) or endpoint titers (right panel). Pearson correlation coefficients are indicated (r values) along with significance (p values). D-E) Glomerular cellularity (D; glomerular mesangial nuclei counts were performed by counting nuclei in 10 random glomerular corpuscles and expressed as average nuclei per glomerulus) and mesangial proliferation (E; glomerular basement membrane thickening or mesangial expansion using a 0-4 scoring system [0 = inapparent, 1 = <50% of glomeruli, segmental; 2 = >50% of glomeruli, segmental; 3 = <50% of glomeruli, predominantly diffuse; and 4 = >50% of glomeruli, predominantly diffuse]) was determined by analysis of periodic-Schiff stained 3 μM thick formalin-fixed paraffin-embedded kidney sections (at least 20 random glomeruli were assessed per section). Age-matched wild type and C4-/- groups are as described for (B).