Association of HLA haplotype with alopecia areata in Chinese Hans

BACKGROUND: Some studies have shown discrepancies in human leucocyte antigen (HLA) associated with alopecia areata (AA) between different ethnic populations. OBJECTIVE: To investigate whether HLA-I, -DQA1 and -DQB1 alleles and the HLA haplotype are associated with AA, and the correlation between the HLA haplotype profile, age of onset and severity of AA in Chinese Hans. METHODS: The polymerase chain reaction-sequence specific primer (PCR-SSP) method was used to analyse the frequencies of HLA class I, -DQA1 and -DQB1 alleles in 192 patients with AA and 252 controls in Chinese Hans. The linkage disequilibrium was calculated using the 2 x 2 table. RESULTS: The 24 two-locus haplotypes [including A*02-B*18, A*02-B*27, A*02-B*52, A*02-Cw*0704, A*02-DQA1*0104, A*02-DQB1*0604, A*02-DQB1*0606, B*18-Cw*0704, B*18-DQA1*0104, B*18-DQA1*0302, B*18-DQB1*0606, B*27-Cw*0704, B*27-DQA1*0104, B*27-DQA1*0302, B*52-Cw*0704, B*52-DQA1*0104, B*52-DQA1*0302, B52-DQB1*0606, Cw*0704-DQA1*0104, Cw*0704-DQA1*0302, Cw*0704-DQB1*0606, DQA1*0104-DQB1*0604, DQA1*0104-DQB1*0606, DQA1*0302-DQB1*0606 (P<0.05)] were associated with AA, while eight extended haplotypes (A*02-B*18-DQA1*0104, A*02-B*27-DQA1*0104, A*02-B*52-DQA1*0104, A*02-B*52-DQA1*0302, A*02-B*52-DQB1*0606, B*52-Cw*0704-DQA1*0104, B*52-Cw*0704-DQA1*0302, A*02-B*52-DQA1*0302-DQB1*0606) were found to be related to AA in Chinese Hans. Through stratified analysis, we found that the extended haplotype B*52-Cw*0704-DQA1*0302 was related to early onset of AA, and no haplotype was only associated with severe AA. CONCLUSION: This is the first detailed report to elucidate HLA haplotypes associated with AA and that demonstrates the significant HLA haplotypes in Chinese Hans AA. The haplotype B*52-Cw*0704-DQA1*0302 was identified to be related to early onset of AA. Our results provide some information for future research on predisposing genes in HLA regions in Chinese Hans.     

The association between endothelin-1 gene polymorphisms and susceptibility to vitiligo in a Korean population

BACKGROUND: Vitiligo is a skin disorder affected by genetic, environmental, local and endocrine factors. Endothelin-1, which is expressed by keratinocytes, has paracrine effects on melanocytes, influencing their homeostasis, proliferation and pigmentation. It is thought to play a role in the skin response to 311-nm, narrow-band ultraviolet irradiation. OBJECTIVE: To investigate the association of endothelin-1 gene (EDN1) polymorphisms with vitiligo in a Korean population. METHODS: To evaluate the expression of endothelin-1 in cultured human keratinocytes after irradiation with narrow-band ultraviolet B (NBUVB), we performed RT-PCR and ELISA. In addition, we genotyped 312 vitiligo patients and 313 matched-healthy controls, and compared the genotype, allele and haplotype frequencies of EDN1 polymorphisms (intron 4 G/A, rs2071942 and exon 5 G/T, rs5370) between the two groups, using PCR-restriction fragment length polymorphism. The effects of sex, onset age, the presence of autoimmune diseases, family history and clinical type were analysed statistically. RESULTS: NBUVB induced the expression of endothelin-1 in cultured human keratinocytes. The genotype distributions and allele frequencies of EDN1 polymorphisms did not differ significantly between vitiligo patients and healthy controls. Moreover, the results were not related to sex, onset age, the presence of autoimmune diseases or family history. Interestingly, the haplotype frequencies of EDN1 polymorphisms differed significantly between vitiligo patients and healthy controls. When analysed according to clinical type, the haplotype frequencies in the focal and segmental clinical types differed significantly from healthy controls. CONCLUSION: This study suggests that EDN1 is related to the development of vitiligo in the Korean population.     

北方汉族银屑病家系HLA基因复合体相关研究

目的研究与中国北方汉族银屑病存在基因易感性的HLAⅠ类基因的分布,并分析与银屑病相关的HLA易感单倍型。方法将核心家系成员分为银屑病患者组(27例)和对照组(9例),应用序列特异性引物-聚合酶链反应扩增HLAⅠ类等位基因。HLA各位点等位基因频率以及基因型频率分布采用直接计数法计算,并做Hardy-Weinberg吻合度检验。两组间的等位基因频率的比较采用四格表x~2检验,HLA各位点间连锁不平衡关系应用连锁不平衡参数计算法。结果在两组受检者中,发现HLA-B*37、HLA-Cw*0602在患者中分布频率高于对照组,是该病的致病基因,HLA-B*48在患者组中分布频率低于对照组,是该病的保护基因。发现在银屑病患者组中有显著性连锁不平衡的HLA单倍型9条。结论证实了HLA基因复合体与银屑病之间存在密切关联。     

Reduction of skin-homing cytotoxic T cells (CD8+ -CLA+) in patients with vitiligo

Background: Vitiligo is a frequently acquired, hereditary disease, characterized by achromic macules due to the absence of melanocytes. In contrast with earlier studies, in which the main pathogenic role was attributed to anti‐melanocyte antibodies, recent papers have emphasized a role for CD8⁺ cytotoxic T lymphocytes in melanocyte destruction. Fifteen percent of peripheral T cell express cutaneous lymphocyte‐associated antigen (CLA), responsible for skin‐homing T cell. Phototherapy is used to treat patients with generalized vitiligo and it has been shown to interfere with CLA⁺ T cells in other skin diseases. Objective: To describe peripheral blood T cell subpopulations' frequency and ability to express the skin‐homing molecule (CLA) in patients with non‐segmental vitiligo, before and after photochemotherapy (PUVA). Patients and Methods: Twenty‐two patients with generalized and active spreading vitiligo were submitted to 30 PUVA‐8MOP sessions. Lymphocyte immunophenotyping was performed by flow cytometry using anti‐CD3, anti‐CD8 and anti‐CLA monoclonal antibodies. Fifteen healthy volunteers, sex‐ and age‐matched, were included as a control group. Results: CD8⁺–CLA⁺ T cells were significantly reduced in number in untreated vitiligo patients (P=0.008) when compared with control individuals, albeit with a more intense CLA expression (P=0.028). These findings were not altered after PUVA. No significant difference was noticed in CD4/CD8 ratios nor in CD4–CLA⁺ T cell numbers between vitiligo patients and controls, both before and after PUVA. Conclusions: CD8–CLA⁺ T cells are reduced in peripheral blood of patients with non‐segmental vitiligo. This finding may be related to the previously reported increase of CD8⁺ cells in both lesions and perilesional skin of these patients.     

Vitiligo in association with human immunodeficiency virus infection

Autoimmune disorders such as vitiligo have been previously reported in patients with human immunodeficiency virus (HIV) infection. We describe a case of vitiligo in association with HIV where a rising CD4 lymphocyte count due to highly active antiretroviral therapy (HAART) closely correlated with changes in the skin. This effect, to our knowledge, has not been previously observed.     

中国汉族人群白癜风6q27区域易感基因精细定位研究

目的:对中国汉族人群白癜风6q27易感区域进行精细定位研究,以期发现白癜风易感基因。方法:收集2942例白癜风患者和2850名正常对照标本,对6q27易感区域内的24个标签SNP位点进行基因分型。采用Plink 1.07和SPSS19.0软件进行统计分析,同时进行条件回归分析和连锁不平衡分析。结果:(1)SNP位点rs2236313、rs9457258、rs720325、rs968334、rs3093025、rs55802221、rs1012656、rs6930998和rs1331299的等位基因频率在病例组和对照组之间具有显著差异,但是通过条件回归分析结果提示没有新的独立信号存在;(2)连锁不平衡分析结果显示rs2236313、rs9457258、rs720325、rs968334和rs30930255位点间具有中度连锁关系(D'0.7,r20.4)。结论:本文证实既往报道的SNP是标签SNP,根据基因型填补以后的注释结果认为白癜风6q27区域的关联基因为RNASET2基因。     

600例白癜风毛发及指甲变化的分析

目的：了解白癜风毛发和指甲的变化情况。方法：对600例白癜风患者的毛发和指甲变化情况进行临床分析。结果：600例白癜风患者中，80例（13．33％）出现点状白甲，2例出现线状白甲，1例出现双手全指甲营养不良，4例指甲出现点状凹陷，40例（6．67％）出现程度和数量不等的甲横沟、纵嵴或甲分离；385例（64．17％）头部出现数量不等的白发；28例（4．67％）部分眉毛变白，37例（6．17％）部分胡须变白，45例（7．50％）部分阴毛变白，25例（4．17％）部分腋毛变白，31例（5．17％）躯干或四肢部分毳毛变白，而白发区无皮肤变白；554例（92．33％）有白斑区部分毛发变白；41例患者（6．83％）合并斑秃。结论：毛发和指甲可能是白癜风皮肤以外较为常见的受累部位。     

Comparisons of clinical features of HLA‐DRB1*07 positive and negative vitiligo patients in Chinese Han population

Background Human leucocyte antigen (HLA)‐II alleles have been found to be associated with vitiligo in different populations, and several studies also suggested that HLA class II alleles/haplotypes were associated with a different type vitiligo. Of HLA class II alleles, DRB1*07 has consistently shown a positive association with vitiligo in Chinese Han population. Objective To further explore the relationship between DRB1*07 and vitiligo and to evaluate the DRB1*07 effect on the clinical features of vitiligo in Chinese Han population. Methods This study investigated DRB1*07 allele distribution in 1178 unrelated Chinese vitiligo patients and 1743 healthy controls using polymerase chain reaction/sequence specific primer method and observed clinical differences between DRB1*07 positive and DRB1*07 negative patients. Results The analysis of the 1178 cases and 1743 controls revealed a highly association between DRB1*07 allele and vitiligo [odds ratio (OR) = 1.97, P = 2.13 × 10^?17]. DRB1*07 positive patients had early disease onset (OR = 1.49, P = 0.001), higher frequency of family history (OR = 1.44, P = 0.006) compared with DRB1*07 negative patients. Conclusions The DRB1*07 showed significant association with vitiligo in the study population. This study confirmed that DRB1*07 positive patients had some obvious clinical differences from DRB1*07 negative patients in the Chinese Han population.     

An association of twenty-nail dystrophy with vitiligo

A rare association of twenty-nail dystrophy with segmental vitiligo is described in two patients. Vitiligo preceded the nail dystrophy. In both cases, all twenty nails were uniformly affected with the nail plates showing longitudinal striations and loss of luster. Longitudinal nail biopsy revealed a histological picture suggestive of eczematous changes and lichen planus respectively. Intramatrix injections of triamcinolone acetonide into the proximal and lateral nail folds were administered with considerable improvement.     

Expression of transporters associated with antigen processing and human leucocyte antigen class I in malignant melanoma and its association with prognostic factors

BACKGROUND: Low expression of transporters associated with antigen processing (TAP) and human leucocyte antigen (HLA) class I, due to defects in the antigen presentation pathway, is frequently found in human tumours, including malignant melanoma (MM). This immune evasion renders many tumours unrecognizable by the host immune surveillance system and appears to play a role in the clinical course of the tumour, probably because it provides tumour cells with a mechanism to escape cytotoxic T-lymphocyte recognition and destruction. However, the histopathological significance of TAP and HLA class I antigen defects in MM remains unclear. OBJECTIVE: To study the expression of TAP and HLA class I antigen in MM and the relationship between them. To investigate the correlation between histopathological characteristics and expression of these molecules in MM. METHODS: Tissue sections from 77 patients with MM and 20 with naevi were examined using immunohistochemistry and morphological quantitative analysis for protein expression of TAP1, TAP2 and HLA class I antigen. RESULTS: Positive TAP1, TAP2 and HLA class I antigen immunostaining was observed in 23%, 12% and 64% of MM lesions, respectively, and the expression of HLA class I was positively correlated with that of TAP1 and TAP2. However, expression of these molecules was positive in all of the pigmented naevi lesions. Reduced TAP1 and TAP2 protein expression in melanoma lesions was significantly associated with invasive growth, Clark's level and tumour-infiltrating lymphocytes. Reduced HLA class I antigen protein expression was only associated with tumour-infiltrating lymphocytes. CONCLUSIONS: Our data suggest that reduced TAP1, TAP2 and HLA class I antigen protein expression in MM may contribute to the immune escape phenotype of human melanoma cells, and the main cause of reduced HLA class I expression may be the decreased TAP1 and TAP2 levels.     

华东地区汉族HLA-Ⅱ类基因与白癜风临床特征的相关性

目的：探讨华东地区汉族HLA-Ⅱ类基因与白癜风临床特征的相关性。方法：采用聚合酶链反应-序列特异性寡核苷酸探针(PCR-SSOP)方法检测华东地区汉族白癜风患者HLA-DRBl、DQAl和DQBl位点的等位基因，使用SPSS 10．0统计软件分析。结果：HLA-DQAl ^*0103与伴发内分泌疾病的患者有显著正相关，DRB l^*09与C4降低的患者明显负相关。按临床类型分析，发现局限型与DRBI*09和DQAl*03正相关，非节段型与DQAl*03正相关。结论：在华东地区汉族人群中，白癜风的临床特征与HIA-Ⅱ类基因有一定的相关性。     

白癜风相关黑素细胞膜抗原VIT150、VIT90、VIT75纯化及分析

目的：纯化和分析白癜风相关的黑素细胞膜抗原，为蛋白质微量测序及筛选、克隆黑素细胞膜抗原打下基础。方法：培养高纯度的正常人黑素细胞，活细胞ELISA法和蛋白免疫印迹法(Western blot)检测并筛选抗黑素细胞的高滴度IgG抗体血清，生物素标记可溶性黑素细胞膜抗原，加入筛选血清与蛋白A-琼脂糖凝胶(protein A-sepharose)进行免疫共沉淀．沉淀后的抗原抗体复合物行平极电泳及电转印，碱性磷酸酶标记的亲合素进行化学发光法检测及鉴定。结果：活细胞ELISA法和Western blot检测结果中共筛选了10例白癜风患者高滴度IgG血清，免疫沉淀、免疫印迹、化学发光法检测后发现10例患者均有阳性条带，其抗原相对分子质量为150000、90000、75000、60000。结论：白癜风患者血清中存在抗黑素细胞膜抗原的自身抗体，抗原相对分子质量主要为150000、90000、75000，通过免疫共沉淀初步纯化了150000、90000和75000抗原。     

HLA‐E*0103X is associated with susceptibility to Pemphigus vulgaris

Non‐classical human leucocyte antigen‐E (HLA‐E) mediates natural killer and CD8+ T‐cell activity, suggesting a role in the regulation of autoimmunity. HLA‐E*0103X/*0103X has been associated with Behcet's disease and HLA‐E *0101/*0103X with childhood onset diabetes. We investigated HLA‐E allele status in 52 Caucasian and Ashkenazi Jewish Pemphigus vulgaris (PV) patients and 51 healthy controls by restriction fragment length polymorphism–polymerase chain reaction and amplification refractory mutation system. Associations were determined via chi‐square test, Fisher's exact test and logistical regression analysis. HLA‐E outcomes included presumed homozygous *0101/*0101 or *0103X/*0103X genotype status or *0101/*0103X heterozygous status. PV did not significantly associate with either *0101/*0101 or *0101/*0103X genotypes. HLA‐E*0103X/*0103X (presumed homozygote) is significantly increased in patients with PV versus controls (P = 0.0146, OR = 3.730, 95%CI = 1.241–11.213). Our data provide the first evidence that HLA‐E*0103X is a marker for genetic risk in PV.     

中国北方汉族泛发型白癜风与HLA-DRB1等位基因的相关

目的:探讨HLA-DRB1等位基因与中国北方汉族泛发型白癜风的相关性。方法:采用聚合酶链反应-序列特异引物(PCR-SSP)技术检测34例北方汉族泛发型白癜风患者的HLA-DRB1等位基因。结果:与262例正常对照组相比较,泛发型白癜风患者HLA-DRB1*0701/02、DRB1*1201/02基因频率显著增高(Pc<0.0001),HLA-DRB1*0901、DRB1*11基因频率降低(但经校正后Pc>0.05);有明确家族史的患者HLA-DRB1*1201/02基因频率显著增高(Pc<0.0001);无家族史者HLA-DRB1*0701/02基因频率显著升高(Pc<0.0001),DRB1*1201/02基因频率显著增高(经校正后Pc>0.05),DRB1*0901基因频率降低(经校正后Pc>0.05)。结论:中国北方汉族人群,HLA-DRB1*0701/02、DRB1*1201/02等位基因可能与泛发型白癜风的发病有关,而DRB1*0901、DRB1*11等位基因可能是防止其发病的“保护因子”,为进一步揭示泛发型白癜风的易感基因及免疫遗传发病机制提供线索。     

Stress in patients with alopecia areata and vitiligo

OBJECTIVE: To study the involvement of stress before the onset/development of alopecia areata and vitiligo. PATIENTS AND METHOD: Forty-five outpatients with alopecia areata and 32 outpatients with vitiligo were enrolled. The design was a case-control study (controls had skin diseases unrelated to stress). Stressful events were evaluated using Holmes and Rahe's social readjustment rating scale. RESULTS: Mean age was around 30 years in both conditions. More than 65% of cases (both alopecia areata and vitiligo) experienced stressful events compared to 22% of controls. The odds ratio was 7.75 for alopecia areata and 6.81 for vitiligo. There was a significant difference in the mean number of stressful events between alopecia areata patients and controls (P = 0.005), and also a significant difference in the number of stressful events between men (P = 0.05) and women (P = 0.001) across these two groups. In the vitiligo group there was a significant difference in the mean number of stressful events between patients and controls only in women (P = 0.02). A potential stressful situation occurred more often in both patient groups. Alopecia areata patients described family problems in 45.6% of patients (especially women), which was statistically significant when compared to controls (P = 0.0004). Personal problems were reported by 35.7% of alopecia areata patients (P = 0.04 compared to controls). Vitiligo patients mentioned personal problems in 47% of cases (one-third were related to exams) and 31% of cases were related to job/financial problems. Again, this was statistically significant when compared to controls (P = 0.0002). CONCLUSIONS: Stress seems to play an important role in the onset and aggravation of both alopecia areata and vitiligo, mostly with one stressful event before disease onset.     

Long-term results of 2-mm punch grafting in patients with vitiligo vulgaris and segmental vitiligo: effect of disease activity

Background Punch grafting is a simple and frequently used technique for the treatment of stable vitiligo, resistant to medical therapy. However, studies reporting long‐term results are exceptional. Objectives To evaluate the long‐term results of 2‐mm punch grafting in patients with vitiligo vulgaris and segmental vitiligo. Methods We studied a prospective cohort study involving 61 patients (25 male, 36 female) with vitiligo vulgaris and nine patients (all male) with segmental vitiligo who underwent 2‐mm punch grafting more than 3 years ago. The main outcome measure was the degree of repigmentation of a single transplanted lesion as measured with a digital image analysis system with a mean follow‐up of 5·2 years. Results In patients with vitiligo vulgaris, 17 lesions (28%) showed excellent, 14 lesions (23%) showed good, 14 lesions (23%) showed fair and 16 lesions (26%) showed poor repigmentation. In patients with segmental vitiligo, seven of nine lesions (78%) showed excellent repigmentation. A cobblestone‐like effect was observed in 19 of 70 patients (27%). Disease activity after punch grafting was reported in 94% of patients with poor repigmentation but in only 18% of patients with excellent repigmentation (χ² test, P < 0·0005). Patients who reported disease activity after transplantation had a lower mean repigmentation than those who did not report disease activity (77% vs. 39%, P < 0·05). Conclusions Two‐millimetre punch grafting in vitiligo is an effective surgical procedure with long‐lasting effect. To prevent a cobblestone‐like effect, we advise the use of smaller grafts (1–1·2 mm). Disease activity after grafting, localization and type of vitiligo, prior ultraviolet B treatment and a Koebnerized donor site influence the long‐term outcome of punch grafting and should be taken into account in the selection of patients eligible for this treatment.     

白癜风患者生活质量评估量表的研究进展

白癜风虽不危及患者生命安全，但疾病的迁延不愈、反复发作给患者造成巨大的心理压力，妨碍患者的人际交往，使患者生活质量明显下降。采用生活质量量表评估患者生活质量是研究疾病对患者影响的常用方法。本文将对白癜风患者生活质量评估量表的应用进展加以综述。