FIGURE 11.

HDI reduce autoantibodies, immunopathology, disease symptoms, and extend lifespan of MRL/Fas^lpr/lpr mice. (A-E) Female MRL/Fas^lpr/lpr mice were started on HDI-water or untreated water at the age of 6 wk, and analyzed at the age of 17 wk. (A) Rostral and dorsal images show characteristic lupus of skin lesion, including a “butterfly” rash. (B) Kidney pathology. The representative photomicrographs of kidney sections stained with hematoxylin and eosin (H & E, left) or FITC-labeled rat mAb to mouse IgG1 and IgG2a (right); scale bars, 100 μm. Data are from one of three independent experiments yielding similar results. (C) ANA visualized by indirect immunofluorescence on HEp-2 cells that were incubated with sera (1:40 dilution) from the MRL/Fas^lpr/lpr mice using FITC-labeled rat mAb to mouse IgG1 and IgG2a; scale bars, 20 μm. Data are from one of three independent experiments yielding similar results. (D) Skin lesion score in the MRL/Fas^lpr/lpr mice (n = 15; 11 of 15 untreated mice, but only 4 of 15 HDI-treated mice, developed skin lesions). p values, unpaired t-test. (E) Titers of circulating anti-dsDNA IgG1 and IgG2a. p values, unpaired t-test. (F-I) Fifty female MRL/Fas^lpr/lpr mice were on untreated water throughout their lives; 15 female MRL/Fas^lpr/lpr mice were on untreated water for the first 6 wk of life, at which age they were started on HDI-water; 15 female MRL/Fas^lpr/lpr mice were on untreated water for the first 17 wk of life, at which age they were showing active disease (skin lesions, proteinuria, anti-dsDNA autoantibodies) and were started on HDI-water. (F) Dorsal images showing severe skin lesions in four female MRL/Fas^lpr/lpr mice before (top, 17 wk of age) and after 4 wk of HDI administration (bottom, 21 wk of age). (G) Skin lesion scores in 15 female MRL/Fas^lpr/lpr mice before (17 wk of age) and after 4 wk of HDI administration (21 wk of age). p values, paired t-test. (H) anti-dsDNA IgM, IgG, IgG1 and IgG2a autoantibody titers in female MRL/Fas^lpr/lpr mice before (17 wk of age) and after 4 wk of HDI administration (21 wk of age). p values, paired t-test. (I) Survival of the 50 female MRL/Fas^lpr/lpr mice given untreated water throughout their lives (black line); the 15 female MRL/Fas^lpr/lpr mice started on HDI-water at 6 wk of age (blue line), and the 15 female MRL/Fas^lpr/lpr mice started on HDI-water at 17 wk of age when they showed already active disease (green line).