The aim of present study was to evaluate CD4(+) /CD8(+) ratio and CD4(+) CD25(hi) FoxP3(+) Tregs in GV patients with reference to their effect on disease onset and progression. Flow cytometry was used for determination of CD4(+) /CD8(+) ratio and Tregs in 82 patients and 50 controls. CD8(+) T-cell counts were significantly higher in GV patients as compared with controls (p = 0.003). Active GV patients showed higher CD8(+) T-cell counts compared with stable GV patients (p = 0.001). The CD4(+) /CD8(+) ratio decreased significantly in patients as compared with controls (p = 0.001). Moreover, the ratio in active GV patients significantly lowered as compared with stable GV patients (p = 0.002). Significant decrease in Treg cell percentage and counts in GV patients was observed compared with controls (p = 0.009, p = 0.008) with significant reduction in FoxP3 expression (p = 0.024). Treg cell percentage and counts were significantly decreased in active GV patients compared with stable GV patients (p = 0.007, p = 0.002). Our results suggest that an imbalance of CD4(+) /CD8(+) ratio and natural Tregs in frequency and function might be involved in the T-cell mediated pathogenesis of GV and its progression.

One of the major hypotheses in the pathogenesis of vitiligo is the oxidative stress hypothesis. Pollution plays a major role in the production of free radicals. Gujarat, a highly industrialized state in India has a high prevalence of vitiligo patients. No previous studies were done on the age-dependent antioxidant status of vitiligo patients in Baroda city, Gujarat. Blood samples were collected from vitiligo patients of different age groups (5-15, 16-25, 26-35, 36-45 yr) and from age matched healthy volunteers. Antioxidant enzymes in blood such as catalase, superoxide dismutase, glutathione peroxidase and non-enzymatic antioxidants such as reduced glutathione and plasma vitamin E were estimated. Lipid peroxidation levels in erythrocytes and the reducing equivalent system, i.e. glucose-6-phosphate dehydrogenase were also measured. Significant increase in superoxide dismutase activity and lipid peroxidation levels in erythrocytes was observed in all age groups of vitiligo patients as compared with age-matched healthy controls, wherein an increase of 55% (P < 0.02) was observed in superoxide dismutase activity and lipid peroxidation levels in 36-45 yr age group. Whole blood glutathione levels, erythrocyte glutathione peroxidase and glucose-6-phosphate dehydrogenase activity were decreased significantly, whereas erythrocyte catalase activity and plasma vitamin E levels were not different in vitiligo patients as compared with age-matched healthy controls. No specific age group showed a significant difference. This is the first report on the age-dependent antioxidant status of vitiligo patients in Baroda. The disease affects individuals of any age group as shown in this study and systemic oxidative stress might precipitate the pathogenesis of vitiligo in susceptible patients.

Figure S1. (a-c) Specificity of the antibody was verified in sections of paraffin-embedded HEK cells transiently transfected with human b) and , respectively. The sections were subjected to immunofluorescence; (a) HEK cells transfected with human IL-20 stained with polyclonal rabbit anti-IL-20 and detected with Alexa 488 (green); (b) HEK cells transfected with human IL-20 and stained with a preimmune serum control from the rabbit in which the polyclonal rabbit anti-IL-20 was generated and detected with Alexa 488 (green); (c) HEK cells transfected with human IL-19 were stained with polyclonal rabbit anti-IL-20, and the nuclei were marked with Hoechst (blue). Stained for IL-20 (green); (d-g) localization of IL-20 protein in lesional psoriatic skin (LS). Paraffin-embedded punch biopsies from LS were subjected to immunofluorescence. (d) Stained for IL-20 (green); (e) same tissue as (d) stained with pre-immune-rabbit serum; (f) stained for IL-20 (green) and nuclear DNA (blue); (g) same tissue as (f), the anti-IL-20 has been pre-absorbed with antigen (IL-20), also stained for nuclear DNA (blue). Figure S2. Immunofluorescence stainings for IL-20 and double immunofluorescence stainings for IL-20 and selected CD markers in untreated lesional psoriatic skin. Figure S3. Effect of treatment with calcipotriol (10 patients, mild-to-moderate psoriasis) left panel, or cyclosporin A (10 patients, moderate-severe psoriasis) right panel, before (day 0) and after treatment for 14 and 28 days and compared with non-lesional skin also at day 0.Data S1. Experimental design. Table S1. Antibodies used for staining paraffinembedded punch biopsies. Abstract: Vitiligo is an acquired depigmentation disorder characterized by the loss of functional melanocytes from the epidermis. Two major theories of vitiligo pathogenesis include autoimmunity and oxidative stress-mediated toxicity in melanocytes. The present study aimed to evaluate both the hypotheses in vitiligo patients and to investigate their role in the disease onset and progression. Antimelanocyte antibody levels and lipid peroxidation (LPO) levels were evaluated in 427 patients and 440 controls; antithyroid peroxidase (TPO) antibody levels were estimated in 102 patients and 72 controls. Patients showed a significant increase in LPO and antimelanocyte antibody levels compared to controls. Antimelanocyte antibody and LPO levels were higher in active vitiligo compared to stable. Only 9.8% of patients showed the presence of anti-TPO antibodies in their circulation. Oxidative stress may be the initial triggering event to precipitate vitiligo in Gujarat population, which is exacerbated by contributing autoimmune factors together with oxidative stress.

No abstract

Our results suggest that these polymorphisms of the IL4 gene may be genetic risk factors for susceptibility towards vitiligo and the upregulation of the IL4 transcript, protein and IgE levels in individuals with susceptible haplotypes reveal the crucial role of IL-4 in the pathogenesis of vitiligo.