Vitiligo is a disease in which melanocytes are selectively destroyed. The disease is thought to be an autoimmune process being there are antibodies to pigment cells in the sera of patients and animals with vitiligo. In the present study, sera from vitiligo patients were examined for reactivity with the human melanoma cell line, SK-Mel-28, by Western blot analysis of solubilized membrane antigens of these cells to identify the pigment cell antigens defined by antibodies in the patients with vitiligo. Antibody reactivity to human melanoma cells (SK-Mel-28) was investigated in 14 patients with vitiligo, and 16 with normal control individuals. Antibodies to the 116-113, 60, 40 KD antigens were associated with vitiligo being present in 79%, 86%, and 43% respectively of the patients with vitiligo, but in only 6%, 38% and 6% of the normal controls. In contrast, antibodies to the 160-155, 78 and 64 KD antigens were equally common in vitiligo and in normal individuals. The results suggest that autoreactivity to pigment cells occurs more commonly in patients with vitiligo than in the normal control and high autoreactivity to pigment cells in the vitiligo sera might be an impertinent epiphemenon to destroyed pigment cell.
Antibodies, Neoplasm/*blood ; Antigens, Neoplasm/immunology ; Autoantibodies/blood ; Blotting, Western ; Human ; Melanoma/*immunology ; Vitiligo/*immunology

To more fully define the nature of the antibody response to melanocytes which is associated with vitiligo, a Western immunoblot assay was used to test the sera of 28 patients with vitiligo (21 with active non-segmental, and 7 with stable segmental diseases) and 26 normal individuals for antibodies to antigens in detergent extracts of melanocyte membrane fractions. Antibodies to melanocytes were found in 26 (93%) of the patients with vitiligo, and in 16 (62%) of the control individuals. Patients with vitiligo and control individuals both had antibodies to an 80 approximately 83 kD antigen. The patient with vitiligo, in addition, had antibody responses to antigens with MWs of 45, 65, and 110 kD. Antibodies to these antigens were present in 46, 25, and 31% of vitiligo patients, but in only 19%. 0%, amd 0%, respectively, of the normal individuals. The heterogeneity of the antibody responses to melanocytes in vitiligo was further confirmed by the presence of antibodies to at least 3 distinct antigens in one-third of vitiligo patients but in none of the normal individuals. There was no difference in antibody response between patients with generalized and segmental vitiligo, suggesting that the pathogenesis of diseases was similar in both cases.
Antibodies/*analysis ; Antigens/immunology ; Blotting, Western ; Human ; Melanocytes/*immunology ; Reference Values ; Support, Non-U.S. Gov't ; Vitiligo/*immunology/*pathology

OBJECTIVETo construct vitiligo-specific HLA-A*0201-peptide tetramers and to apply the constructed tetramers in detection of vitiligo-specific cytotoxic T lymphocytes (CTL).

METHODSProteins HLA-A0201*-BSP and β2M were obtained by effective prokaryotic expression. The purified proteins were refolded with vitiligo antigen peptides MelanA 26-35, gp100 209-217, and tyrosinase 1-9, respectively to form HLA-A*0201-peptide complex. The complex was biotinylated by BirA enzyme and purified by gel-filtration chromatography. The tetramers were generated by mixing the complex with phycoerythrin (PE)-streptavidin at a ratio of 4∶1 and identified by Dot-blot assay. The capacity of tetramer to detect vitiligo-specific CTL was analyzed by flow cytometry.

RESULTSThe biotinylation of vitiligo-specific HLA-A*0201-peptide tetramers were successfully performed by Dot-blot. Flow cytometry analysis indicated that the tetramer effectively bound to specific CTL from peripheral blood of patients with vitiligo.

CONCLUSIONThree kinds of biotinylated vitiligo-specific HLA-A*0201-peptide tetramers have been constructed successfully. The tetramer can detect antigen specific CTL from patients with vitiligo.

Biotinylation ; Flow Cytometry ; HLA-A2 Antigen ; Humans ; Peptides ; T-Lymphocytes, Cytotoxic ; cytology ; Vitiligo ; diagnosis ; immunology

OBJECTIVETo detect the serum levels of melanocyte antibodies and explore the relation between melanoma antigen recognized by T-cells (MART-1) and vitiligo in children.

METHODSThe serum samples were collected from children with vitiligo to test the autoantibodies, and divided into low- and high-titer group according to the test results. Melanocytes were incubated with the serum samples, and the changes of melanocyte surface antigen were evaluated using specific MART-1 antibody.

RESULTSThe serum melanocyte antibody levels in children with vitiligo were significantly higher than those in normal subjects. The expression level of melanocyte surface antigen MART-1 increased obviously after incubation of the melanocyte with high antibody titer serum samples, and MART-1 was found to specifically bind to specific MART-1 antibody.

CONCLUSIONMelanocytes MART-1 may correlate to the autoimmune mechanism in children with vitiligo.

Adolescent ; Autoantibodies ; blood ; Child ; Child, Preschool ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; MART-1 Antigen ; immunology ; Male ; Melanocytes ; immunology ; T-Lymphocytes ; immunology ; Vitiligo ; immunology

Vitiligo is frequently associated with autoimmune diseases, such as multiple glandular insuffi ciencies and thyroid diseases. In addition, various circulating antiorgan antibodies are found in patients with vitiligo. This raises the possibility that vitiligo might also be an antibody associated au toimmune disease. Variou. alterations in peripheral mononuclear cells, especially T-cells and T-cell subsets have been desiribed in patients with vitiligo. The discovery of circulating antimelanocyte antibodies in patients with vitiligo demonstrateci that vitiligo may be associted with alterations in the specific immunity to melanocytes. These vit iligo antibodies, which are more common in patients with vitiligo than in normal individuals, react with cell surface pigment cell antigens with MWs of approximately 150, 90, 75, 40-45, and 35 kDa, and can kill rnelanocytes in vitro. It has been suggested tiat melanocytes are much more sensitive to toxic or immune mediatece injury that other cutaneou; cell types, thus explaining their apparently selective destruction in vitiligo despite the rather bro d specificity of these vitiligo antibodies. However vitiligo autoantibodies are not found in all vitilio patients. Some of t,hem are present in patients without vitiligo. Tak ing into account the common occurrence of circulation autoantibodies irrelevant to the pathogene sis of the cutaneous hypomelanosis in vitiligo patients, the pathogenetic role of these vitiligo anti bodies has not yet been demonstrated, and the possibility that they represent an impertineni epiphenomenon in vitiligo cannot be ruled out.
Allergy and Immunology* ; Antibodies ; Autoantibodies ; Autoimmune Diseases ; Humans ; Hypopigmentation ; Melanocytes ; Sensitivity and Specificity ; T-Lymphocyte Subsets ; T-Lymphocytes ; Thyroid Diseases ; Vitiligo*

Vitiligo is an acquired, progressive depigmenting disorder of unknown etiology. In this study, to clarify pathogenesis of vitiligo, the marginal skin of actively spreading and stable vitiligo was examined using ICAM-1, HLA-DR, CD4 and CD8 monoclonal antibodies. In immunohistochemical study, ICAM-1 was expressed in four of five epidermis in active lesions, but not in stable lesion. Dermal ICAM-1 was also expressed in all active and stable lesions. HLA-DR was also expressed in all active epidermis in active lesions, but two of five epidermis in stable lesion. Dermal HLA-DR was also expressed in all active and stable lesion. CD4 lymphocytes were expressed more strongly in active lesion, but CD8 lymphocytes were not different in both lesions. There was no significant difference of degree of positivity with CD4 and CD8 in normal control specimens. In conclusion, we think that ICAM-1 and HLA-DR expression, cytokines released from keratinocytes, melanocytes or lymphocytes and infiltration of activated T-lymphocytes play an important role in disease activity.
Antigens, CD4/metabolism ; Antigens, CD8/metabolism ; Comparative Study ; HLA-DR Antigens/metabolism ; Human ; Immunohistochemistry ; Intercellular Adhesion Molecule-1/metabolism ; Skin/immunology ; Vitiligo/*immunology

OBJECTIVETo observe the clinical efficacy of Zengse Pill ( ZSP) on patients with vitiligo of qi-stagnancy and blood-stasis syndrome type (V-QB), and to preliminarily explore its mechanism of action.

METHODSSixty-five V-QB patients, with their diagnosis confirmed by clinical examination, were randomized by digital table method into two groups, with 31 patients in the control group and 34 in the treatment group. Cobamamide (2 tablets) was administered orally to all patients, and Psoralea tincture (a self-formulated preparation) was applied externally thrice a day. In addition, for patients in the treatment group, ZSP was given orally, at 5 pills per dose, 3 times every day. The therapeutic course for both groups was 3 months. Patients were re-examined every half-month, and changes in the skin lesions were observed and recorded. The levels of lymphocyte subsets, serum immune globulin, and complement C3 and C4 in patients were determined before and after the therapeutic course and compared with the corresponding indexes determined in 21 healthy subjects.

RESULTSThe total effective rate in the treatment group was 82.4%, which was markedly higher than that in the control group (54.8%), showing a significant difference (P<0.05). After treatment, CD(4) (+) percentage, CD(4) (+)/CD(8) (+) ratio, and blood levels of C3 and C4 increased, while CD(8) (+) percentage decreased in the treatment group (P<0.05 or P<0.01). All these indexes remained unchanged in the control group, and the respective comparisons between groups showed significant differences (P<0.01).

CONCLUSIONZSP has a definite clinical effect on the treatment of V-QB but with no evident adverse reactions, and it can increase the CD(4) (+) percentage, CD(4) (+)/CD(8) (+) ratio, and the levels of serum C3 and C4, thus regulating the immunity of the organism, which might be one of its mechanisms of action.

Adolescent ; Adult ; CD4-CD8 Ratio ; Complement C3 ; analysis ; Complement C4 ; analysis ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Immunoglobulins ; blood ; Lymphocyte Subsets ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Qi ; Vitiligo ; drug therapy ; immunology

PURPOSE: Vitiligo prevalence and its associated comorbidities rate have been reported variably among different populations. We aimed to determine the prevalence of vitiligo in Korea along with the baseline rate of comorbidities and compared the risks to the general population using hospital visit information of the total population in Korea. MATERIALS AND METHODS: We assessed demographic characteristics of vitiligo patients in Korean population from 2009 to 2011 in a nationwide data from Health Insurance Review Assessment Service. Patients who had at least one visit to Korea's primary, secondary, or tertiary referral hospitals with International Classification of Diseases, 10th Revision, Clinical Modification diagnosis code for vitiligo were identified. As a supplementary study, comorbidities associated with vitiligo were selected for further review to calculate relative risks compared to the general population. RESULTS: The annual prevalence of vitiligo determined by hospital-visiting rate in Korea was 0.12% to 0.13% over a three year period. In sync with other previous epidemiological studies, there was bimodal distribution among the age groups and no difference between genders. Also, vitiligo in Korean population was associated with various autoimmune/non-autoimmune diseases such as thyroiditis, atopic dermatitis, and psoriasis. CONCLUSION: This study was by far the most comprehensive review on prevalence of vitiligo using a data of total population in Korea. The prevalence is within a range of those reported in previous literatures, and increased risk of comorbidities such as thyroid diseases and psoriasis in vitiligo might aid clinicians in the initial work up of vitiligo patients and concurrent follow ups.
Adult ; Aged ; Autoimmune Diseases/*epidemiology/immunology ; Comorbidity ; Diabetes Mellitus, Type 1/epidemiology ; Female ; Humans ; Male ; Middle Aged ; Population Surveillance ; Prevalence ; Republic of Korea/epidemiology ; Socioeconomic Factors ; Thyroid Diseases/epidemiology ; Vitiligo/*epidemiology