Generalized Vitiligo Patients Research Articles

Evaluation of IL-38, a Newly-introduced Cytokine, in Sera of Vitiligo Patients and Its Relation to Clinical Features.

Vitiligo is thought to be an autoimmune disorder caused by melanocytes dysfunction and depigmentation. Among different executors of the immune system in developing the disease, the role of various cytokines has been defined. We have focused on IL-38, the tenth member of IL-1 cytokine family with a proposed anti-inflammatory role, which has not hitherto been introduced as an anti-inflammatory factor in vitiligo. Sixty-nine generalized vitiligo patients and 72-year-old- and sex-matched healthy individuals were included in this study. IL-38 level was evaluated in sera of all participants using ELISA assay. The relation of IL-38 level to patients characteristics was evaluated. IL-38 serum level in vitiligo patients (159.5±39.7 pg/ml) was lower than the healthy controls (166.7±34.8pg/ml) (P = 0.039). A weak negative correlation between the age of male patients and their IL-38 serum levels was identified (r = 0.38, P = 0.058). Evaluation of the IL-38 serum levels relationship with patients clinical characteristics showed no correlation with disease onset, stage of depigmentation, and disease activity status. The lower levels of IL-38 as an anti-inflammatory cytokine support the inflammatory nature of vitiligo. It indicates the difference of IL-38 in sera of vitiligo patients and healthy controls, as the first report of the lower level of this cytokine in the context of vitiligo. The reason of this difference remains to be clarified; as there are not sufficient study reports revealing the role of gender, ethnicity and inflammation on the cytokine network in the context of vitiligo.

HLAB27 may confer protection to COVID-19 in generalized vitiligo patients from South Gujarat population

Coronavirus disease 2019 (COVID-19) is caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), while generalized vitiligo(GV) is an autoimmune disease that causes the loss of functional melanocytes, resulting in white patches all over the body. Human Leukocyte Antigen (HLA) plays a crucial role in immune response to pathogens. Studies assessing the link between GV and COVID-19 are lacking; therefore, our current study was aimed to establish the association between GV and HLAB27 by genotyping the HLAB27 allele in 150 GV patients and 150 controls from South Gujarat population through polymerase chain reaction-sequence-specific primers (PCR-SSP) method. Additionally, we assessed the correlation of GV with COVID-19 and the influence of HLAB27 on COVID-19 development. Interestingly, our study suggested that the HLAB27 allele was prevalent in GV patients as compared to controls (52% vs 35.33%; p = 0.0051). Moreover, the occurrence of COVID-19 was significantly lower in GV patients than in controls (10% vs 32.66%; p < 0.0001). Disease activity-based analysis suggested that COVID-19 occurrence was significantly lower in active vitiligo (AV) patients as compared to stable vitiligo (SV) patients(6.87% vs 31.57%; p = 0.0045). Furthermore, COVID-19 development was significantly reduced in HLAB27 positive individuals as compared to HLAB27 negative individuals (p = 0.0025). Overall, our study suggests, for the first time, that HLAB27 allele might be a genetic risk factor for GV susceptibility, and an ongoing immune response in GV patients, more specifically in AV patients, might protect against COVID-19 infection in South Gujarat population. Additionally, our study highlighted the likely role of HLAB27 in protection against COVID-19 development.

Altered Regulatory T cell-mediated Natural Killer Cells Suppression may lead to Generalized Vitiligo

Generalized vitiligo(GV) is characterized by white patches due to autoimmune loss of melanocytes. Regulatory T cells(Tregs) maintain immune homeostasis, while NK cells eliminate pathogens and tumors. Increased NK cell frequency and reduced Treg frequency and suppressive capacity are observed in vitiligo patients. However, studies assessing Treg-mediated suppression of NK cell functions in GV are lacking. Therefore, our study aimed to assess in vitro Treg-mediated suppression of NK cells function over K562 and SK-Mel-28 cells in 31 GV patients and 30 controls using the BrdU-cell proliferation assay. We found decreased Treg-mediated suppression of NK cell function in GV patients(p=0.0289). Moreover, increased NK cell-mediated K562 and SK-Mel-28 cells’ suppression was observed in GV patients(p=0.0207,p=0.0419). Disease activity-based analysis, suggested reduced Treg-mediated suppression of NK cell function and increased NK cell function in active vitiligo patients(p=0.03,p=0.0436). Interestingly, age-based analysis suggested decreased Treg-mediated suppression of NK cell function in 1-20 and 21-40 years age groups compared to 41-60 years age group of GV patients(p=0.005,p=0.0380). Overall, our study, for the first time, suggests that decreased Treg-mediated suppression of NK cells may lead to increased destruction of melanocytes in GV, and this knowledge may help in developing effective therapeutics based on Tregs and NK cells for GV.

Compromised melanocyte survival due to decreased suppression of CD4+ & CD8+ resident memory T cells by impaired TRM-regulatory T cells in generalized vitiligo patients.

Regulatory T cells (Tregs) are involved in the suppression of activated T cells in generalized vitiligo (GV). The study was aimed to investigate resident memory (TRM)-Tregs and antigen-specific Tregs' numbers and functional defects in 25 GV patients and 20 controls. CD4+ & CD8+ TRM cell proliferation was assessed by BrDU assay; production of IL-10, TGF-β, IFN-γ, perforin and granzyme B were assessed by ELISA and enumeration of TRM cells was done by flowcytometry. GV patients showed significantly increased frequency and absolute count of CD4+ & CD8+ TRM cells in lesional (L), perilesional (PL) and non-lesional (NL) skin compared to controls (p = 0.0003, p = 0.0029 & p = 0.0115, respectively & p = 0.0003, p = 0.003 & p = 0.086, respectively). Whereas, TRM-Treg (p < 0.0001 & p = 0.0015) and antigen-specific Tregs (p = 0.0014 & p = 0.003) exhibited significantly decreased frequency and absolute counts in L & PL skin. GV patients showed reduced suppression of CD8+ & CD4+ TRM cells (with increased IFN-γ, perforin & granzyme B) and decreased TRM-Tregs and antigen-specific Tregs (with decreased IL-10 & TGF-β production) and reduced proliferation of SK-Mel-28 cells in co-culture systems. Immunohistochemistry revealed increased expression of TRM stimulating cytokines: IL-15 & IL-17A and reduced expression of TGF-β & IL-10 in L, PL, NL skins compared to controls. These results for the first time suggest that decreased and impaired TRM-Tregs and antigen-specific Tregs are unable to suppress CD4+ & CD8+ TRMs' cytotoxic function and their proliferation due to decrease production of immunosuppressive cytokines (IL-10 & TGF-β) and increased production of TRM based IFN-γ, perforin and granzyme B production, thus compromising the melanocyte survival in GV.

Genetic association of nuclear factor of activated T cells’ 3′UTR and structural polymorphisms with susceptibility to generalized vitiligo in Gujarat population

Generalized vitiligo (GV) is an autoimmune skin depigmenting disease characterized by loss of functional melanocytes. Nuclear factor of activated T cells (NFATs) play a key role in regulatory T cells’ (Tregs) activation and function. Our previous studies have highlighted the role of reduced NFATs expression and activity in impaired Tregs suppressive capacity, leading to GV pathogenesis. 3′UTR region and structural single nucleotide polymorphisms(SNPs) could lead to reduced NFAT expression and activity. Therefore, we studied the association of NFATs 3′UTR [NFATC2 rs4811198 (T > G) &NFATC4 rs11848279 (A > G)] and structural [NFATC1 rs754093 (T > G) &NFATC2 rs12479626 (T > C)] SNPs in 427 GV patients and 415 controls from Gujarat population by Polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP). Additionally, we carried out genotype-phenotype correlation and in silico analysis to assess the effect of NFATs SNPs on NFATs expression and structure. NFATC2 rs4811198 (T > G) 3′ UTR &NFATC2 rs12479626 (T > C) structural SNPs were significantly associated with GV (p < 0.0001). Interestingly, for NFATC2 rs4811198 (T > G) SNP, there was a significant difference in the TT vs GG genotypes’ frequencies (p = 0.0034; Table 2), and for NFATC2 rs12479626 (T > C) SNP there was a significant difference between TT vs TC and CC genotypes’ frequencies (p < 0.0001 & p = 0.0002) between GV patients and controls. Furthermore, Odds ratio suggested that the susceptible alleles for NFATC2 rs4811198 (T > G) &NFATC2 rs12479626 (T > C) SNPs increased the risk of GV by 1.38 & 3.04 fold. However, the NFAT 3′ UTR [NFATC2 rs4811198 (T > G)] and structural [NFATC1 rs754093 (T > G)] SNPs were not significantly associated with GV. Interestingly, the genotype-phenotype correlation suggested that the susceptible ‘G’ allele of NFATC2 rs4811198 (T > G) &NFATC4 rs11848279 (A > G) 3′ UTR SNPs lead to reduced NFATC2 and NFATC4 expression (p < 0.0001). Furthermore, in silico analysis suggested that hsa-miR-3183 & hsa-miR-6720-3p miRNAs specifically bound to ‘G’ allele of NFATC2 rs4811198 SNP and has-miR-4652-3p miRNA specifically bound to ‘G’ allele of NFATC4 rs11848279 SNP. Overall, our study suggests that NFATC2 rs4811198 (T > G) 3′ UTR &NFATC2 rs12479626 (T > C) structural SNPs may be associated with GV susceptibility in Gujarat population. Moreover, the susceptible alleles for the 3′ UTR SNPs could lead to reduced NFATs levels, which may further possibly, affect the Treg suppressive function leading to GV.

Decreased GZMB, NRP1, ITPR1, and SERPINB9 Transcripts Lead to Reduced Regulatory T Cells Suppressive Capacity in Generalized Vitiligo Patients

Generalized vitiligo (GV) is an autoimmune skin disease characterized by bilateral white patches over the entire body. Regulatory T cells (Tregs) maintain peripheral tolerance; however, they are found to be reduced in numbers and function in vitiligo patients. The exact mechanism for reduced Treg suppressive capacity is unknown. Therefore, we aimed to assess transcript levels of Tregs-associated immunosuppressive genes (GZMB, NRP1, PDCD1, FASLG, and TNFRS18), regulatory molecules of Tregs suppressive function (SERPINB9, ITPR1, and UBASH3A), and Treg-associated transcription factors (GATA2, GATA3, RUNX1, STAT3, and STAT5) in 52 GV patients and 48 controls by real-time PCR (qPCR). We found significantly reduced GZMB, NRP1, SERPINB9, and ITPR1 transcripts in GV Tregs compared to controls (p = 0.03, p = 0.023, p = 0.0045, and p < 0.0001, respectively). There were 0.44-, 0.45-, 0.32-, and 0.54-fold decrease in GZMB, NRP1, SERPINB9, and ITPR1 transcripts in GV Tregs. Additionally, disease activity and severity-based analyses revealed significantly decreased GZMB (p = 0.019 and 0.034), SERPINB9 (p = 0.031 and p = 0.035), and ITPR1 (p = 0.0003 and p = 0.034) transcripts in active vitiligo and severe GV patients' Tregs. Interestingly, we found a positive correlation for ITPR1 with GZMB (r = 0.45, p = 0.0009) and SERPINB9 (r = 0.52, p = 0.001) transcripts in GV Tregs. Moreover, we found positive correlation for percentage Treg mediated suppression of CD4+ and CD8+T cells with ITPR1 (r = 0.54; r = 0.49), GZMB (r = 0.61; r = 0.58), NRP1 (r = 0.55; r = 0.52), and SERPINB9 (r = 0.56; r = 0.48) in GV Tregs. Further, calcium treatment of Tregs resulted into significantly increased ITPR1, SERPINB9, and GZMB transcripts in GV Tregs (p = 0.023, p = 0.0345, p = 0.02). Overall, our results for the first time revealed the crucial role of GZMB, NRP1, SERPINB9, and ITPR1 transcripts in decreased Treg suppressive capacity leading to GV pathogenesis, progression, and severity. In addition, our study highlighted that ITPR1 might be linked with decreased GZMB and NRP1 expression in GV Tregs. Moreover, our study for the first time suggest that increased SERPINB9 transcripts may lead to endogenous granzyme B-mediated Tregs apoptosis, and calcium treatment of Tregs may improve the Treg suppressive capacity. These findings may further aid in development of Treg-based therapeutics for GV.

Calcium controlled NFATc1 activation enhances suppressive capacity of regulatory T cells isolated from generalized vitiligo patients.

NFATs and FOXP3 are linked with impaired regulatory T-cells (Tregs) in generalized vitiligo (GV). To elucidate calcium mediated NFATc1 signalling pathway and its effect on Treg suppressive capacity in GV. Calcium levels, calcineurin, NFATc1 and GSK-3β activity and cell proliferation were assessed in 52 GV patients and 50 controls by calcium assay kit, calcineurin phosphatase assay kit, TransAM NFATc1 kit, GSK-3β ELISA and BrdU cell proliferation assay. Transcripts (CNB, CAM, GSK3B, DYRK1A and calcium channel genes) and protein (IFN-γ, IL-10 and TGF-β) expressions were assessed by qPCR and ELISA, respectively. Reduced plasma and intracellular Tregs calcium levels and ORAI1 transcripts suggested altered calcium homeostasis in GV Tregs (p=0.00387, p=0.0048, p < 0.0001), which led to decreased calcineurin and NFATc1 activity in GV Tregs (p=0.0299, p < 0.0001). CNB and CAM transcripts were reduced in GV Tregs (p < 0.0001, p=0.0004). GSK-3β activity, GSK3B and DYRK1A transcripts significantly increased in GV Tregs (p=0.0134, p < 0.0001 and p < 0.0001). Plasma (p=0.0225, p=0.032) and intracellular Treg (p=0.0035, p=0.005) calcium levels, calcineurin (p=0.001) and NFATc1 (p=0.001, p < 0.0001) activity and ORAI1 (p=0.0093, p < 0.0001), CAM and CNB (p=0.0214) transcripts significantly decreased in active vitiligo (AV) and severe GV (sGV) Tregs. Calcium treatment significantly increased intracellular calcium and ORAI1 transcripts in GV Tregs (p=0.0042, p=0.0035). Moreover, calcium treatment enhanced calcineurin and NFATc1 activity in GV Tregs (p=0.0128, p < 0.0001). Remarkably, calcium treatment increased Treg mediated suppression of CD4+ and CD8+ T-cells (p=0.015, p=0.006) in GV and increased Tregs associated cytokines: IL-10 (p=0.0323, p=0.009), TGF-β (p=0.0321, p=0.01) and decreased IFN-γ production (p=0.001, p=0.016) by CD4+ and CD8+ T-cells. Intracellular calcium levels positively correlated with calcineurin (r=0.83; p < 0.0001) and NFATc1 (r=0.61; p < 0.0001) activity, suggesting the enhanced Treg immunosuppressive capacity after calcium treatment. Our study for the first time suggests that reduced plasma calcium and ORAI1 transcripts are linked to calcium uptake defects in Tregs, which leads to reduced calcineurin and NFATc1 activation, thereby contributing to decreased Tregs immunosuppressive capacity in GV. Elevated GSK-3β activity and GSKB and DYRK1A transcripts are involved in reduced NFATc1 activity in GV Tregs. Overall, the study suggests that calcium-NFATc1-signalling pathway is likely to be involved in defective Tregs function and can be implicated for development of effective Treg mediated therapeutics for GV.

Aktif ve Generalize Vitiligolu Hastalarda Azalmış Oksidatif Stres Belirteçleri

Objective&#x0D; Oxidative stress has been shown to play a role in the pathophysiology of several diseases, making it a popular yet contentious research area. There is some evidence that selective melanocyte destruction may have developed in vitiligo patients as a result of elevated oxidative stress. The purpose of this study is to investigate the impact of oxidative stress on lipid, protein, and nucleic acid metabolism in vitiligo patients.&#x0D; Method&#x0D; We used ELISA method to measure serum oxidative stress markers in patients with generalized vitiligo who had newly formed lesions in the previous three months but had not been treated, as well as healthy controls. Malondialdehyde (MDA), 2,4-dinitrophenyl hydrazone (DNPH), 8-hydroxy-2'-deoxyguanosine (8-OHdG), and uncoupling protein 2 (UCP2) levels were measured to assess the influence of reactive oxygen derivatives on lipid, protein, nucleic acid metabolism, and mitochondria, respectively. &#x0D; Results&#x0D; The study included 84 participants, including 64 active generalized vitiligo patients and 20 healthy controls with similar age and gender distribution. In the serum of vitiligo patients, we detected significantly lower levels of MDA (ng/mL, mean±SD=12±19; 33.4±35.9), DNPH (ng/mL, mean±SD=2±3.1; 6±7.4), 8-OHdG (ng/mL, mean±SD=11.7±17.9; 32.7±37) and UCP2 (ng/mL, mean±SD=8.7±13.7; 21.5±28.4).&#x0D; &#x0D; Conclusion&#x0D; Although there is significant evidence that oxidative stress plays a role in the pathophysiology of vitiligo, the studies should be interpreted cautiously due to the heterogeneity in the methodology, complexity of the oxidative stress pathways, and potential publication bias. Large-scale studies using a standardized methodology are required to determine how significant oxidative stress is in the core pathophysiology of vitiligo and which pathways it primarily affects.

A pilot study of combined oral minocycline and narrowband UVB phototherapy in vitiligo: A randomized, double-blind, placebo-controlled trial.

Narrowband ultraviolet B (NBUVB) phototherapy is an effective therapeutic option for generalized vitiligo. Previous reports showed the potential benefit of minocycline to stop disease progression in vitiligo. Meanwhile, minocycline has antioxidative, anti-inflammatory, and immunomodulating properties. There is no clinical study combining oral minocycline and NBUVB for treating generalized vitiligo. This study aims to compare the efficacy and safety of the combination treatment of NBUVB plus oral minocycline with NBUVB alone in generalized vitiligo. A randomized, double-blinded, placebo-controlled pilot study was conducted. Patients were randomly treated with either combined oral minocycline 100 mg per day plus NBUVB phototherapy or placebo plus NBUVB. All patients recieved NBUVB two times per week, for 12 weeks. The outcomes were assessed using Vitiligo Area Scoring Index score (VASI) percent change, quartile grading scale (QGS) of repigmentation, and Vitiligo Disease Activity Index (VIDA) score. Fourteen generalized vitiligo patients were included, and seven cases were assigned in each group. At week 12, the mean VASI score was decreased by 28.87% (24.15) in the minocycline group compared to 27.26% (7.98) in placebo group (p= 0.886). No significant difference was observed between both treatment modalities in QGS of repigmentation and mean VIDA score change. Two of the seven patients (29%) receiving minocycline developed hyperpigmentation, dark-brown and muddy brown discoloration, which was only confined to some vitiliginous patches. In conclusion, combination therapy with oral minocycline does not enhance the efficacy of NBUVB in generalized vitiligo. Due to the high incidence of drug-induced skin hyperpigmentation, minocycline plus NBUVB should be avoided.

Differential Expression and in Silico Functional Analysis of Plasma MicroRNAs in the Pathogenesis of Non-segmental Vitiligo.

Vitiligo is a disease characterized by acquired depigmentation, white macules, and patches on the skin due to the dysfunction of epidermal melanocytes. In this study, we attempt to profile the microRNA (miRNA) expression patterns and predict the potential targets, assessing the biological functions of differentially expressed miRNAs in the blood of generalized vitiligo patients. Peripheral blood samples were taken from all participants, and the expression levels of 89 identified miRNAs were analyzed with real-time quantitative polymerase chain reaction (PCR). The results indicated significant upregulation of six miRNAs and downregulation of 19 miRNAs in the plasma of vitiligo patients. The top three upregulated miRNAs were hsa-miR-451a, hsa-miR-25-3p, and hsa-miR-19a-3p, and the top three downregulated miRNAs were hsa-miR-146a-5p, hsa-miR-940, and hsa-miR-142-3p. Moreover, the miRNA expression profiles of patients with Type 3 and Type 4 phototypes were substantially different in such a way that the patients with Type 3 phototype would be more prone to the emergence of melanoma and cancer. While significant variations in the expression patterns of miRNAs in male and female vitiligo patients were demonstrated, miR-let-7i-5p, miR-19a-3p, miR-25-3p, and miR-451a were commonly upregulated, and miR-142-3p and miR-146a-5p were commonly repressed in both sexes. This study may shed light on the roles of differentially expressed miRNAs in vitiligo patients by examining the miRNA expression patterns and the combined effects of miRNA and their predicted targets.

Levels of TNF-α, IL-6, IL-17, IL-37 cytokines in patients with active vitiligo

Objectives Vitiligo is an autoimmune disease, and its pathogenesis involves changes in cytokine levels in the affected patients. Tumor necrosis factor-alpha (TNF-α), interleukin (IL)-6, and IL-17 from pro-inflammatory cytokines, IL-37 in a recently detected anti-inflammatory activity. The aim of our study was to determine serum TNF-α, IL-6, IL-17, IL-37 levels in patients with vitiligo to understand their possible roles in the disease etiology and to compare the results with the healthy controls. Methods The study included 48 generalized vitiligo patients who were diagnosed with vitiligo, had an increase in the lesions within the last 3 months, and did not receive any systemic or topical treatment during this period; furthermore, 18 healthy controls were included. Results Patient group: n = 48, mean age = 30.48 ± 9.86 years; control group: n = 18, mean age = 28.27 ± 9.66 years. Individuals in the patient group had significantly higher serum levels of IL-37(t = 3.90, p < .001), IL-6 (t = 3.39, p < .05), IL-17 (t = 2.08, p < .05), and TNF-α (t = 4.69 p < .001) than in the control group. Conclusion The high levels of (pro-anti) inflammatory cytokines in vitiligo patients draw attention to the importance of cytokines in the pathogenesis of the disease.

Association of FOXP3 and GAGE10 promoter polymorphisms and decreased FOXP3 expression in regulatory T cells with susceptibility to generalized vitiligo in Gujarat population.

Alterations in regulatory T (Treg) cells have been observed in generalized vitiligo (GV) patients and decreased Forkhead Box P3 (FOXP3) has been implicated in the disease pathogenesis. The present study examined FOXP3 rs3761547(A>G), rs3761548(C>A), rs2232365(A>G) and GAGE10 rs11798415(A>T) promoter single nucleotide polymorphisms (SNPs) in 419 GV patients and 429 controls from Gujarat population using PCR-RFLP and ARMS-PCR. Real-time PCR and flow cytometry were used for assessment of FOXP3 mRNA and protein levels respectively in 96 GV patients and 90 controls. The frequency of genotypes (p<0.001) and alleles (p=0.012 & p=0.040) for rs3761547(A>G) and rs11798415(A>T) SNPs significantly differed between GV patients and controls. FOXP3 mRNA and protein levels were significantly decreased (p<0.001) in GV Tregs compared to controls. Active vitiligo (AV) Tregs showed significantly reduced FOXP3 mRNA and protein levels compared to that of stable vitiligo (SV) (p=0.02 & p=0.039). The correlation of genotype-phenotype of FOXP3 SNPs suggested reduced FOXP3 mRNA (p=0.019, p<0.001 & p<0.001) and protein (p=0.028, p<0.001 & p=0.022) levels in patients with susceptible GG, AA and GG genotypes respectively. The GAGT, GCGT & ACGT haplotypes were prevalent in GV patients (p=0.004, p=0.004 & p=0.016); however, GAGT & GCGT were overrepresented in patients with AV (p=0.044 & p=0.024). The susceptible GAGT and GCGT haplotypes in patients exhibited reduction in FOXP3 mRNA (p=0.014 & p=0.019) and protein (p=0.024 & p=0.028). DNA-protein docking analysis revealed reduced binding for transcription factor C/EBP to the susceptible allele 'G' (rs3761547) compared to A allele. For the first time, the study suggests significant association of FOXP3 rs3761547(A>G) & GAGE10 rs11798415(A>T) SNPs with susceptibility to GV in Gujarat population. In addition, the likely role of these SNPs in altered FOXP3 expression of Tregs may possibly affect Treg suppressive function in GV.

Decreased suppression of CD8+ and CD4+ T cells by peripheral regulatory T cells in generalized vitiligo due to reduced NFATC1 and FOXP3 proteins.

Regulatory T cells (Tregs) are involved in the suppression of activated T cells in generalized vitiligo (GV). The study was aimed to investigate Tregs functional defects in Treg:CD8+ and Treg:CD4+ T cells' co-culture systems of 55 GV patients and 45 controls. CD8+ and CD4+ T-cell proliferation was assessed by BrdU assay; production of IL-10, TGF-β and IFN-γ cytokines was assessed by ELISA; and FOXP3, CD25, NFATC1 and CD44 proteins were measured by flow cytometry. Generalized vitiligo patients showed reduced suppression of CD8+ and CD4+ T cells (P=.0384, P=.0084), increased IFN-γ (P<.0001, P=.0019), decreased IL-10 and TGF-β (P<.0001) and decreased FOXP3, CD25 and NFATC1 proteins (P<.0001). Active vitiligo (AV) patients showed reduced suppression of CD8+ & CD4+ T cells (P=.006, P=.015), increased IFN-γ (P=.036, P=.045), decreased IL-10 (P=.009, P=.021), FOXP3 (P=.0244) and NFATC1 (P=.019). Severe GV (50%-75% VASI) patients showed reduced suppression of CD8+ and CD4+ T cells (P=.0003, P=.001), increased IFN-γ (P=.0029, P<.0001), decreased IL-10 (P=.0057, P=.0017), FOXP3 (P=.002) and NFATC1 (P=.0347). VASI score was positively correlated with the suppression of CD8+ and CD4+ T cells (P=.0006, P<.0001), IL-10 (P=.0096, P=.029), FOXP3 (P=.0008) and NFATC1 (P=.043), whereas it was negatively correlated with IFN-γ (P=.0029, P=.0017). Early age of onset patients' Tregs demonstrated decreased suppression of CD8+ and CD4+ T cells (P=.0156, P=.0074), decreased TGF-β (P=.0212, P=.0083) and NFATC1 (P=.0103). NFATC1 was positively correlated with FOXP3 in Tregs (P<.0001). Our results suggest impaired Tregs suppressive function in GV patients due to decreased NFATC1, FOXP3, CD25, IL-10 and TGF-β resulting into increased CD8+ and CD4+ T-cell proliferation and IFN-γ production. For the first time, decreased NFATC1 levels were correlated with decreased FOXP3, thereby altering Treg cell function in GV patients. Additionally, decreased Treg cell function also affected onset, activity and severity of GV.

Altered expression of nuclear factor of activated T cells, forkhead box P3, and immune‐suppressive genes in regulatory T cells of generalized vitiligo patients

The study was aimed to analyze expression of nuclear factor of activated T cells (NFATs), forkhead box P3 (FOXP3), and their associated genes (sCTLA4, flCTLA4, IL10, TGFB, IL2, IL4, CD25) in regulatory T cells (Tregs) of 48 generalized vitiligo (GV) patients and 45 unaffected controls. The transcripts of NFATC1 to NFATC4, FOXP3, IL10, flCTLA4 (p<.0001), NFAT5 (p=.0003), sCTLA4 (p=.001), and FOXP3 protein in Tregs and plasma IL-10 levels were reduced significantly (p<.0001) in GV Tregs compared to controls. The FOXP3 promoter polymorphisms [rs3761548(C>A), rs3761547(A>G), and rs2232365(A>G)] revealed significantly decreased FOXP3 protein levels in patients' Tregs with susceptible AA, GG, and GG genotypes (p<.0001, p=.028, p=.022, respectively). The active vitiligo Tregs showed reduced levels of NFATC3, NFATC4, NFAT5, FOXP3, TGFB, and flCTLA4 transcripts (p=.0005, p=.0003, p=.0002, p=.020, p<.0001, p=.006, respectively) and FOXP3 and TGF-β proteins (p=.0394 and p=.0013) compared to stable vitiligo. Early-onset patients (1-20years) demonstrated decreased IL-10, sCTLA-4, flCTLA-4, TGFB, and FOXP3 transcripts and FOXP3 protein as compared to late-onset patients (41-60years) (p=.001, p=.003, p=.009, p=.005, p=.038, p=.0226, respectively). Overall, our results for the first time suggest a likely role of NFATs and FOXP3 together with Treg immune-suppressive genes in GV pathogenesis and disease progression, warranting additional investigations.

The role of IFIH1 gene rs1990760 and rs2111485 single-nucleotide polymorphisms in generalized vitiligo predisposition

Background/aim Interferon-induced helicase (IFIH1) is a gene locus that has been recently defined as a candidate for susceptibility to generalized vitiligo (GV). The objectives of this study were to assess the association of IFIH1 gene, rs2111485, and rs1990760 single-nucleotide polymorphisms (SNP) with susceptibility to GV and the autoimmune diseases accompanying GV.Materials and methods We prospectively studied GV patients and frequency-matched healthy controls by age and sex. The genotypes of the participants were determined for rs1990760 and rs2111485 SNPs of IFIH1. Dominant, recessive, and additive models were evaluated for each SNP adjusted for age and sex. Results The patients and their controls were observed to be in the Hardy–Weinberg equilibrium for SNP1 (2q24.2, rs1990760, IFIH1, T/C) and SNP2 (2q24.2, rs2111485, IFIH1, G/A), respectively (all P > 0.7). For SNP1, every T allel addition was significantly associated with 1.53 times protectiveness in terms of vitiligo risk (P = 0.033). As for SNP2, every G allel addition was associated with 1.42 times protectiveness, close to statistical significance (P = 0.100). Conclusions: We detected that for SNP1, each T allel and for SNP2, each G allel are protective in terms of vitiligo development. Hereby, we confirmed that IFIH1 gene locus has a role in GV susceptibility.

Dynamic thiol/disulfide homeostasis in serum of patients with generalized vitiligo

Vitiligo is a multifactorial disorder commonly associated with hypo-/depigmentation in the skin and may influence both children and adults psychologically because of the notable leopard-skin-like appearance. This study was designed to investigate the thiol/disulfide homeostasis in patients with generalized vitiligo and to determine its relationship with some of the demographical characteristics. Seventy-six generalized vitiligo patients and 67 healthy individuals were included in the study. Blood serum native thiol, disulfide and total thiol concentrations, together with some hematological parameters, were determined. Results demonstrated that native and total thiol contents, and their ratios, were significantly lower in vitiligo patients. Disulfide/native thiol and disulfide/total thiol ratios were significantly higher in the patient group. Progressivity of the disease strongly regulated the thiol/disulfide homeostasis in such a way that active vitiligo patients had reduced native and total thiol levels but increased disulfide/native thiol and disulfide/total thiol ratios. Moreover, there was a statistically significant negative correlation between both duration of the vitiligo and native and total thiol contents. As these results clearly demonstrated that thiol/disulfide homeostasis was shifted toward disulfide formation in patients with generalized vitiligo, determining the dynamic nature of thiol/disulfide homeostasis can be used to monitor disease progression.

Plasma & urinary catecholamines & urinary vanillylmandelic acid levels in patients with generalized vitiligo.

Background & objectives:Vitiligo is an acquired skin disease characterized by depigmented areas of the skin. Increased release of catecholamines from autonomic nerve endings in microenvironment of melanocytes in affected skin might be involved in the aetiopathogenesis of vitiligo. Levels of catecholamines are considered as being related to onset or worsening of the disease. Therefore, in this study, the role of catecholamines was evaluated in mapping disease stability and outcome of vitiligo patients undergoing melanocyte transfer.Methods:In this study, circulatory and urinary levels of catecholamine (CA) and vanillylmandelic acid (VMA) were determined in 45 individuals (30 vitiligo patients and 15 healthy controls) using ELISA.Results:A significant increase for plasma and urinary catecholamines along with VMA was observed as compared to healthy controls. When the pre- and post-intervention levels were analyzed in responders and non-responders, respectively, only dopamine showed significant decline in urine, rest of the molecules in plasma as well as urine showed non-significant decline except VMA which showed insignificant increase.Interpretation & conclusions:Levels of plasma/urinary epinephrine, and plasma dopamine, could not be established as biomarkers for disease stability or successful outcome of autologous melanocyte transfer in generalized vitiligo patients. However, dopamine (urine) might be of help in determining the stability in patients with generalized vitiligo undergoing melanocyte transfer. Further studies need to be done on a large sample of patients to confirm our findings.

Association of HLA-DRB1*1201/02, DRB1*0701/02, DQA1*0302 and DQB1*0303 Alleles with population Uygur patients of vitiligo

Vitiligo is associated with different human leukocyte antigens (HLAs), but the relationship of HLAs and Uygur population people is unknown. The study aimed to investigate whether there is an association between HLADRB1* 1201/02, DRB1*0701/02, DQA1*0302, and DQB1*0303 and the incidence of vitiligo in a Chinese Uygur population and further evaluate their effect on the clinical features of vitiligo. A total of 308 Chinese Uygur patients with vitiligo and 310 healthy controls were included in the study. HLA-DRB1*1201/02, DRB1*0701/02, DQA1*0302 and DQB1*0303 allele distribution was investigated by polymerase chain reaction (PCR), the sequence-specific primer method. Family history, clinical types, phase, etc. were evaluated. The data were analyzed by the chi-square test. A significant difference was found in the distribution of HLA-DRB1*1201/02, DRB1*0701/02, DQA1*0302, and DQB1*0303 between the vitiligo patients and healthy subjects in the studied population. A comparison of the clinical features of DRB1*1201/02 (+) and DRB1*1201/02 (-) patients revealed a significant difference between localized and generalized vitiligo patients, (OR: 0.55, 95% CI: 0.31–0.99, P = 0.044). The progressive phase of vitiligo was more common for DRB1*1201-positive patients than for HLA-DRB1*1201/02-negative patients (OR: 3.18, 95% CI: 1.48–6.82, P = 0.002). The other clinical features were similar in both groups. In conclusion, our findings suggest that HLA-DRB1*1201/02, DRB1*0701/02, DQA1*0302, and DQB1*0303 are associated with vitiligo susceptibility, and DRB1*1202/02-positive patients have some obvious clinical differences from DRB1*1202/02-negative patients in the Chinese Uygur population.

Neutrophil to lymphocyte ratio in patients with vitiligo

IntroductionThere are a few studies showing an increased risk of insulin resistance, metabolic syndrome, and oxidative stress in patients with vitiligo.AimTo investigate whether systemic inflammation is increased in vitiligo patients in a case-control study design.Material and methodsNonsegmental vitiligo patients who had been followed at the outpatient dermatology clinic of a university-affiliated teaching hospital, and healthy controls were enrolled in the study. Patients who were receiving systemic treatments and having a systemic disease such as diabetes mellitus and thyroiditis were excluded. Demographic features were recorded and peripheral blood samples were taken from all participants to study serum whole blood count, creatinine, and C-reactive protein (CRP).ResultsFifty patients with localized vitiligo, 43 patients with generalized vitiligo, and 50 healthy volunteers were enrolled in the study. Neutrophil to lymphocyte ratio and serum CRP levels were significantly higher in patients who have generalized vitiligo than those with localized vitiligo and healthy controls. However, there was no significant difference regarding neutrophil to lymphocyte ratio (NLR) and CRP between localized vitiligo and control groups.ConclusionsPatients with generalized vitiligo seem to have increased systemic inflammation compared with localized vitiligo and control subjects in our cohort. To the best of our knowledge, this is the first study in the literature showing increased NLR values in generalized vitiligo patients. Further studies with cardiovascular disease markers are required to elicit this association better.

Investigation of suitable starting doses of narrowband UVB in Asian vitiligo patients: a pilot study.

Phototherapy is the preferred treatment modality for active and generalized vitiligo. One of the widely accepted consensus on starting dose of phototherapy is using a uniform dose of 280 mJ/cm2 regardless of patients' Fitzpatrick skin phototype (SPT). However, in many clinical experiences with Asian vitiligo patients, the protocol seems suboptimal. To gather more evidence on establishing a higher starting dosage for Asian vitiligo patients undergoing phototherapy. We enrolled generalized vitiligo patients with lesions sized adequate enough for phototest. Minimal erythema dose (MED) of vitiligo lesion and non-lesion was measured along with melanin index (MI). Relatively, a wide range of MED and MI was observed even among patients with similar SPT. The range of MED for lesional skin was 300-700 mJ/cm2 and the MED for non-lesion was 500-800 mJ/cm2 . Correlation was noted between lesional MED and non-lesional MI (Spearman correlation, ρ = 0.664 [P < 0.036]) and mean lesional MED was approximately 65% of mean non-lesional MED. Results from phototest and tolerability of patients to doses higher than 280 mJ/cm2 may indicate that higher starting doses might be appropriate for Asian vitiligo patients.