Table 1.
Demographics and clinical features of vitiligo subjects.
Table 2.
Demographics and clinical features of normal control subjects.
Figure 1.
Transcriptome Analysis of Vitiligo and Normal Skin Biopsies.
A heat map is constructed by Gene Spring software (see methods) comparing the relative expression levels of the 30 significantly altered genes in vitiligo skin. Depicted are the expression levels of these genes in individual samples relative to their corresponding expression reference levels, which are the averages of expression in the 16 normal skin biopsies. Red squares: Genes with up-regulation in that sample compared with normal skin of healthy volunteers. Green squares: Genes with down-regulation in that specific sample compared with normal skin of healthy volunteers. Yellow squares: no significant change between the sample and the normal skin of healthy volunteers.
Table 3.
Down-regulated genes in vitiligo skin.
Table 4.
Up-regulated genes in vitiligo skin.
Table 5.
Enriched molecular pathways in vitiligo differentially expressed genes*.
Figure 2.
Explant culture analysis of natural killer cell infiltrates in biopsies of vitiligo lesional and non-lesional skin.
Natural killer (NK) cells from 6 pairs of vitiligo skin explants (lesional and non-lesional) and 5 normal skin explants were cultured on Cellfoam matrices (see methods section) and analyzed using flow cytometry, with the gate set on total live cells A: Skin-resident CD56bright CD3-ve NK cells in normal control skin, vitiligo non-lesional skin and lesional skin by scatter plot. B: Further gating on the CD56bright CD3-ve cells revealed that majority of the NK cells in vitiligo skin were granzyme B-positive. C: Dot plot of all samples analyzed for CD56bright CD3- natural killer cells. The difference in the proportion of resident natural killer cells between normal skin and the respective vitiligo non-lesional and lesional skin is statistically significant (p = 0.0043; mean ± SEM). Comparisons between the respective groups are indicated in the figure by lines with an asterisk (*) denoting statistical significance (p<0.05). Abbreviations: NS: normal skin; NLS: non-lesional skin; LS: lesional-skin.
Figure 3.
Distribution of natural killer cells and cytotoxic T cells in vitiligo lesional and non-lesional skin.
Skin biopsies taken from 12 vitiligo patients and 6 normal individuals were subjected to immunofluorescence analysis of natural killer (NK) cells. A: Micrographs showing natural killer cells (CD3−/NKG2D+) (red) present in vitiligo lesional and non-lesional skin but absent from the normal skin of healthy volunteers. Some NK cells are in close proximity to the basal epidermal layer where melanocytes reside (arrows). In addition, increased numbers of cytotoxic T cells (CD3+/NKG2D+) (yellow: co-localization of red and green) as well as non-cytotoxic T cells (CD3+/NKG2D−) (green) were also found in both vitiligo peri-lesional and lesional skin. B: Quantification of cells demonstrates a statistically significant increase in NK cells, cytotoxic T cells and non-cytotoxic T cells in vitiligo non-lesional skin (p = 0.0021, 0.0015, 0.001; mean ± SEM) and lesional skin (p = 0.021, 0.0017, 0.0023; mean ± SEM) as compared with normal skin. Color keys: Green: CD3 (a pan-T cell marker); Red: NKG2D (NK cell activation receptor); and blue: DAPI (nuclear stain). Comparisons between the respective groups are indicated in the figure by lines with an asterisk (*) denoting statistical significance (p<0.05). Abbreviations: NS: normal skin; NLS: non-lesional skin; LS: lesional-skin. Magnification: 400×; scale bar: 20 µm.