Noncicatrizing alopecias; with special reference to alopecia areata

Since the epochal work of Hamilton there has been general acceptance of the causal relationship of the male sex hormone, age and familial inheritance in development of male pattern baldness. Some of the medicaments used in recent years may cause a diffuse loss of scalp hair. Alopecia that accompanies disease states is probably due to generalized toxemia and disturbances in metabolism. Sometimes male pattern baldness occurs in physiologic states, as exemplified by diffuse hair loss occasionally in the postpartum period. Alopecia areata deserves a critical appraisal, since it may be evidence of underlying neuropsychotic states that need psychiatric diagnosis and treatment. The development of alopecia totalis or universalis in 50 per cent of the prepuberal cases of alopecia areata is of real significance, especially since so very few patients recover their normal scalp hair. The conclusions reached by the authors of two articles reporting on 368 cases of alopecia areata, alopecia totalis and alopecia universalis that the evidence is overwhelming against the malfunction of the endocrine glands as the cause of alopecia areata must be considered real contributions to our understanding of this condition.A few conditions simulate alopecia areata. Probably the ones which are seen most often are trichotillomania and patchy baldness caused by agents used in hair waving and straightening. Our findings in 22 cases of alopecia areata of a persistent inflammatory perivascular and perifollicular infiltrate, massive plugging of the ostia, disappearance of robust hair follicles and diminution in total number of hair follicles and sometimes fibrosis are not necessarily diagnostic of alopecia areata but seem to be very definitely characteristic. Treatment for alopecia areata is of little avail. At this time we do not recommend the general use of the corticosteroids despite the improvement of scalp appearance in the majority of instances in which the systemic administration of these hormones have been employed.     

NONCICATRIZING ALOPECIAS—With Special Reference to Alopecia Areata

Since the epochal work of Hamilton there has been general acceptance of the causal relationship of the male sex hormone, age and familial inheritance in development of male pattern baldness.Some of the medicaments used in recent years may cause a diffuse loss of scalp hair. Alopecia that accompanies disease states is probably due to generalized toxemia and disturbances in metabolism. Sometimes male pattern baldness occurs in physiologic states, as exemplified by diffuse hair loss occasionally in the postpartum period.Alopecia areata deserves a critical appraisal, since it may be evidence of underlying neuropsychotic states that need psychiatric diagnosis and treatment. The development of alopecia totalis or universalis in 50 per cent of the prepuberal cases of alopecia areata is of real significance, especially since so very few patients recover their normal scalp hair.The conclusions reached by the authors of two articles reporting on 368 cases of alopecia areata, alopecia totalis and alopecia universalis that the evidence is overwhelming against the malfunction of the endocrine glands as the cause of alopecia areata must be considered real contributions to our understanding of this condition.A few conditions simulate alopecia areata. Probably the ones which are seen most often are trichotillomania and patchy baldness caused by agents used in hair waving and straightening.Our findings in 22 cases of alopecia areata of a persistent inflammatory perivascular and perifollicular infiltrate, massive plugging of the ostia, disappearance of robust hair follicles and diminution in total number of hair follicles and sometimes fibrosis are not necessarily diagnostic of alopecia areata but seem to be very definitely characteristic.Treatment for alopecia areata is of little avail. At this time we do not recommend the general use of the corticosteroids despite the improvement of scalp appearance in the majority of instances in which the systemic administration of these hormones have been employed.     

Contribution of interleukin 1beta and KM loci to alopecia areata

Alopecia areata is a common skin disease in which proinflammatory cytokines such as IL-1beta may play a pathogenic role. In this study, we examined the distribution of genotypes of an IL-1beta single base change polymorphism at position +3953 in patients with alopecia areata. The distribution of immunoglobulin kappa light chain (KM) genotypes was similarly examined. The data obtained showed that the IL-1beta and KM loci act cooperatively to significantly increase susceptibility to alopecia areata.     

Profiling of alopecia areata autoantigens based on protein microarray technology

Protein biochips have a great potential in future parallel processing of complex samples as a research tool and in diagnostics. For the generation of protein biochips, highly automated technologies have been developed for cDNA expression library production, high throughput protein expression, large scale analysis of proteins, and protein microarray generation. Using this technology, we present here a strategy to identify potential autoantigens involved in the pathogenesis of alopecia areata, an often chronic disease leading to the rapid loss of scalp hair. Only little is known about the putative autoantigen(s) involved in this process. By combining protein microarray technology with the use of large cDNA expression libraries, we profiled the autoantibody repertoire of sera from alopecia areata patients against a human protein array consisting of 37,200 redundant, recombinant human proteins. The data sets obtained from incubations with patient sera were compared with control sera from clinically healthy persons and to background incubations with anti-human IgG antibodies. From these results, a smaller protein subset was generated and subjected to qualitative and quantitative validation on highly sensitive protein microarrays to identify novel alopecia areata-associated autoantigens. Eight autoantigens were identified by protein chip technology and were successfully confirmed by Western blot analysis. These autoantigens were arrayed on protein microarrays to generate a disease-associated protein chip. To confirm the specificity of the results obtained, sera from patients with psoriasis or hand and foot eczema as well as skin allergy were additionally examined on the disease-associated protein chip. By using alopecia areata as a model for an autoimmune disease, our investigations show that the protein microarray technology has potential for the identification and evaluation of autoantigens as well as in diagnosis such as to differentiate alopecia areata from other skin diseases.     

Epitope mapping of human aromatic L-amino acid decarboxylase

Autoimmune polyendocrine syndrome type I (APS I) is a rare hereditary condition considered a model disease for organ specific autoimmunity. A wide range of autoantibodies targeting antigens present in the affected organs have been identified. Autoantibodies against aromatic L-amino acid decarboxylase (AADC) are present in about 50% of APS I patients. In order to increase our understanding of autoantibody specificity in APS I, the aim of the present study was to localize target regions on AADC recognized by sera from APS I patients. Using several complementing strategies, we have shown that autoantibodies against AADC mainly recognize conformational epitopes. The major antigenic determinants were detected N-terminally to amino acid residue 237. Replacement of amino acids 227-230 (ERDK) with alanine residues reduced the reactivity towards AADC by >80% in all patient sera tested, suggesting that amino acids 227-230 are an important part of an immunodominant epitope.     

Effector mechanisms of the autoimmune syndrome in the murine model of autoimmune polyglandular syndrome type 1

Mutations in the Aire gene result in a clinical phenomenon known as Autoimmune Polyglandular Syndrome (APS) Type I, which classically manifests as a triad of adrenal insufficiency, hypoparathyroidism, and chronic mucocutaneous infections. In addition to this triad, a number of other autoimmune diseases have been observed in APS1 patients including Sj?gren's syndrome, vitiligo, alopecia, uveitis, and others. Aire-deficient mice, the animal model for APS1, have highlighted the role of the thymus in the disease process and demonstrated a failure in central tolerance in aire-deficient mice. However, autoantibodies have been observed against multiple organs in both mice and humans, making it unclear what the specific role of B and T cells are in the pathogenesis of disease. Using the aire-deficient mouse as a preclinical model for APS1, we have investigated the relative contribution of specific lymphocyte populations, with the goal of identifying the cell populations which may be targeted for rational therapeutic design. In this study, we show that T cells are indispensable to the breakdown of self-tolerance, in contrast to B cells which play a more limited role in autoimmunity. Th1 polarized CD4(+) T cells, in particular, are major contributors to the autoimmune response. With this knowledge, we go on to use therapies targeted at T cells to investigate their ability to modulate disease in vivo. Depletion of CD4(+) T cells using a neutralizing Ab ameliorated the disease process. Thus, therapies targeted specifically at the CD4(+) T cell subset may help control autoimmune disease in patients with APS1.     

甘草酸二胺肠溶胶囊联合液氮冷冻治疗斑秃的疗效

为了研究观察不同剂量的甘草酸二胺肠溶胶囊联合液氮冷冻治疗斑秃的疗效分析,以及检测甘草酸二胺肠溶胶囊对患者肝肾功能的影响,进一步阐明甘草酸二胺肠溶胶囊对斑秃治疗的效果与对人体的健康影响,本研究以仅使用液氮冷冻治疗斑秃为对照组,设低剂量甘草酸二胺肠溶胶囊联合液氮为低剂量组(50 mg/次),高剂量甘草酸二胺肠溶胶囊联合液氮为高剂量组(200 mg/次),一日3次,共治疗12周,以斑秃皮损面积及严重程度指数(PASI)评分为疗效评价。采用血液生化检验(肝,肾功检查)检测3组治疗前后的丙氨酸氯基转移酶、尿素氮和肌酐水平。研究结果表明,治疗后高剂量组总有效率(92.50%)高于低剂量组(82.50%)及对照组(55.00%)(p<0.05)。高剂量组PASI评分及治疗前后丙氨酸氯基转移酶、尿素氮、肌酐水平相对于低剂量组及对照组无统计学意义(p>0.05)。本研究初步结论表明,高剂量的甘草酸二胺肠溶胶囊联合液氮治疗斑秃疗效显著高于甘草酸二胺肠溶胶囊联合液氮及仅使用液氮冷冻治疗;甘草酸二胺肠溶胶囊安全可靠,不影响患者的肝肾功能。     

Methods of hair loss evaluation in patients with endocrine disorders

Hair loss may accompany several endocrine disorders, including hypopituitarism, hypothyreosis, hyperthyreosis, hypoparathyroidism, diabetes mellitus, growth hormone deficiency, hyperprolactinaemia, polycystic ovary syndrome, SAHA syndrome, congenital adrenal hyperplasia, Cushing syndrome, or virilising tumours. Most patients with endocrine disorders present with diffuse non-scarring alopecia, such as anagen effluvium, telogen effluvium or androgenetic alopecia. Focal non-scarring alopecia, such as alopecia areata coexisting with autoimmune thyroiditis, is less frequent and scarring alopecia is a rare finding in patients with endocrine abnormalities. In some cases an endocrine disorder may be suspected based on dermatological findings during hair loss evaluation. Classic methods of hair evaluation include hair weighing, pull test, wash test, the trichogram, and histopathological examination. Newly developed non-invasive diagnostic techniques include the phototrichogram, trichoscan, trichoscopy, and reflectance confocal microscopy.     

Km1 allotype association with one subgroup of alopecia areata.

The distribution of the immunoglobulin kappa light-chain determinant, Km1, was examined in 52 patients with alopecia areata, 22 of whom possessed serum antibodies to thyroid antigens. The allotype frequency was significantly higher than that found in a referent population of 105 healthy subjects. Furthermore, it was determined that within the patient population, the Km1 allotype was associated with the absence of detectable serum autoantibodies. In such patients, this association may be due to linkage disequilibrium between the chromosome 2 gene coding for Km1 and a chromosome 2 gene coding for susceptibility to alopecia areata.     

Topical minoxidil in the treatment of alopecia areata.

A modified double blind crossover study was performed to assess the effect of 1% topical minoxidil as compared with placebo in 30 patients with alopecia areata and alopecia totalis. The active preparation produced a highly significant incidence of hair regrowth. A cosmetically acceptable response was noted in 16 patients. No side effects were seen. The study confirmed that topical minoxidil will induce new hair growth in alopecia areata but that it is less likely to do so in more severe and extensive disease. Furthermore, patients with alopecia universalis and totalis may not respond at all. Nevertheless, as compared with other drugs minoxidil applied topically is relatively non-toxic, is easy to use, and has no systemic or local side effects.     

HLA-DQA1*0301-associated susceptibility for autoimmune polyglandular syndrome type II and III.

No significant differences were reported for the frequency of DR3-DQ2 and DR4-DQ8 haplotypes in a recent study of one of the largest cohorts worldwide of patients with isolated Addison's disease compared to patients with APS II. However, previous studies had suggested that the HLA-DQ genes, especially DQA1*0102, may be a genetic marker for resistance to autoimmune thyroid disease, which is the most frequent disease in APS II or III. Until now, HLA-DQA1 alleles have not been systematically investigated in APS. We determined the HLA-DR and HLA-DQA1 association in 112 unrelated patients with APS II (n = 29), APS III (n = 83) and 184 unrelated patients with single-component diseases without further manifestations of APS: Graves' disease (n = 70), Hashimoto's thyroiditis (n = 53), autoimmune Addison's disease (n = 15), vitiligo (n = 16) and alopecia (n = 30), and 72 healthy controls - German Caucasians - to identify possible predisposing and protective HLA class II alleles in APS. In agreement with previous studies, we detected a significantly higher frequency of DR 3 and/or DR 4 in patients with APS II and III compared to controls. In patients with APS II, we detected a significantly higher frequency of DQA1*0301 and *0501 compared to controls confirming the increased frequency of an extended HLA DRB1-*04-DQA1-*03-DQB-*03 haplotype as previously described. In contrast, only DQA1*0301 was increased in our patients with APS III compared to controls. Moreover, we detected an increased frequency of DQA1*0301 in patients with APS, whereas DQA1*0301 was only significantly elevated in alopecia in patients with single-component diseases without APS. Therefore, our results indicate an association between DQA1*0301 and APS II or III since this allele was otherwise not significantly associated with any of its component diseases except alopecia. Moreover, our data imply that the allele DQA1*0301 is a marker of increased risk for further APS manifestations in patients who suffer from an organ-specific autoimmune disease.     

A mouse model of clonal CD8+ T lymphocyte-mediated alopecia areata progressing to alopecia universalis

Alopecia areata is among the most prevalent autoimmune diseases, yet compared with other autoimmune conditions, it is not well studied. This in part results from limitations in the C3H/HeJ mouse and DEBR rat model systems most commonly used to study the disease, which display a low frequency and late onset. We describe a novel high-incidence model for spontaneous alopecia areata. The 1MOG244 T cell expresses dual TCRA chains, one of which, when combined with the single TCRB present, promotes the development of CD8(+) T cells with specificity for hair follicles. Retroviral transgenic mice expressing this TCR develop spontaneous alopecia areata at nearly 100% incidence. Disease initially follows a reticular pattern, with regionally cyclic episodes of hair loss and regrowth, and ultimately progresses to alopecia universalis. Alopecia development is associated with CD8(+) T cell activation, migration into the intrafollicular region, and hair follicle destruction. The disease may be adoptively transferred with T lymphocytes and is class I and not class II MHC-dependent. Pathologic T cells primarily express IFNG and IL-17 early in disease, with dramatic increases in cytokine production and recruitment of IL-4 and IL-10 production with disease progression. Inhibition of individual cytokines did not significantly alter disease incidence, potentially indicating redundancy in cytokine responses. These results therefore characterize a new high-incidence model for alopecia areata in C57BL/6J mice, the first to our knowledge to apply a monoclonal TCR, and indicate that class I MHC-restricted CD8(+) T lymphocytes can independently mediate the pathologic response.     

Quantitation of tyrosine hydroxylase, protein levels: spot immunolabeling with an affinity-purified antibody

Tyrosine hydroxylase was purified from bovine adrenal chromaffin cells and rat pheochromocytoma using a rapid (less than 2 days) procedure performed at room temperature. Rabbits were immunized with purified enzyme that was denatured with sodium dodecylsulfate, and antibodies to tyrosine hydroxylase were affinity-purified from immune sera. A Western blot procedure using the affinity-purified antibodies and 125I-protein A demonstrated a selective labeling of a single Mr approximately 62,000 band in samples from a number of different tissues. The relative lack of background 125I-protein A binding permitted the development of a quantitative spot immunolabeling procedure for tyrosine hydroxylase protein. The sensitivity of the assay is 1-2 ng of enzyme. Essentially identical standard curves were obtained with tyrosine hydroxylase purified from rat pheochromocytoma, rat corpus striatum, and bovine adrenal medulla. An extract of PC 12 cells (clonal rat pheochromocytoma cells) was calibrated against purified rat pheochromocytoma tyrosine hydroxylase and used as an external standard against which levels of tyrosine hydroxylase in PC12 cells and other tissue were quantified. With this procedure, qualitative assessment of tyrosine hydroxylase protein levels can be obtained in a few hours and quantitative assessment can be obtained in less than a day.     

PTPN22 gene polymorphism in Egyptian alopecia areata patients and its impact on response to diphencyprone immunotherapy

PTPN22 1858C>T gene polymorphism has been associated with several autoimmune disorders including alopecia areata. The aim of the current study was to investigate the effect of the inherited genetic polymorphism 1858C>T of PTPN22 gene on the predisposition to severe forms of alopecia areata and its effect on the response to DPC treatment. To achieve our aim, PTPN22 1858C>T genotyping was performed by PCR-based restricted fragment length polymorphism (PCR-RFLP) analysis. The study included 103 Egyptian patients with extensive alopecia areata treated by DPC. Hundred healthy age and sex matched blood donors were included in the current study as a control group. Results of genotyping showed that PTPN22 CT and TT mutant genotypes were significantly higher in AA patients compared to controls and conferred increase risk of AA (OR = 2.601, 95% CI = 1.081–6.255). Statistical comparison between AA patients with wild and mutant genotypes revealed that the duration of the illness was significantly longer in those harboring the mutant genotypes. Moreover, the association of other autoimmune diseases as atopy and diabetes mellitus was higher in patients with mutant genotypes. Furthermore, PTPN22 1858C>T genetic polymorphism did not affect the patients' response to DPC immunotherapy.     

1550nm非剥脱点阵激光治疗斑秃的临床随机对照试验

比较1 550 nm非剥脱点阵激光和醋酸曲安奈德注射液皮损内注射治疗斑秃的临床疗效。将72例斑秃患者随机分成两组,治疗组36例、对照组36例,治疗组用1 550 nm非剥脱点阵激光治疗,对照组用醋酸曲安奈德注射液皮损内注射,每2周1次,共4次,12周后观察结果。治疗12周后治疗组治愈29例(80.5%),显效5例,总有效率94.4%;对照组治愈12例(40.0%),显效8例,总有效率66.7%;治疗组有效率和治愈率显著高于对照组(P0.01和P0.01)。治疗结束后6个月,治疗组复发3例(8.8%);对照组复发5例(25%),两组复发率比较,差异无统计学意义(P0.05)。治疗组的不良反应发生率(8.8%)比对照组(25%)小,不良反应发生率差异有显著性(P0.01)。1 550 nm非剥脱点阵激光治疗斑秃疗效显著。     

复方甘草酸苷片联合米诺地尔酊局部治疗斑秃的疗效及其对外周血IFN-γ、IL-10水平的影响

目的：探讨复方甘草酸苷片联合米诺地尔酊局部治疗斑秃的疗效及其对外周血IFN-γ、IL．10水平的影响。方法：选取斑秃患者120例,随机分为观察组与对照组,每组各60例。对照组口服灵丹片,于皮损处外涂米诺地尔酊;观察组在此基础上,再给予复方甘草酸苷片;两组疗程均为3个月,治疗前后观察SALT评分、血清IFN-γ和IL-10水平,评价临床疗效,记录不良反应。结果：治疗后,观察组与对照组总有效率分别为91．67％、71．67％（P〈0．05）,SALT评分均不同程度的降低,且观察组SALT评分低于对照组（P〈0．05）;观察组血清IFN-γ水平降低,IL-10升高（P〈0．05）,且与对照组相应指标比较,差异具有统计学意义（P〈0．05）。观察组与对照组不良反应发生率分别为1．67％、3．33％（P〉0．05）。结论：复方甘草酸苷片联合米诺地尔酊局部治疗斑秃具有较高的临床总有效率,改善了斑秃的皮损状况,调节Th1／Th2细胞因子趋于平衡,且具有较高的安全性,对斑秃的临床治疗具有一定的指导借鉴意义,值得临床推广应用。     

Failure to induce alopecia areata in C3H/HeJ mice with exogenous interferon gamma

Alopecia areata is a cell mediated autoimmune disease that targets actively growing, anagen stage hair follicles in several mammalian species. Upregulation of MHC I due to interferon gamma is considered to be one of the initiating steps. To test this hypothesis we used the spontaneous C3H/HeJ mouse model, induced anagen by wax stripping the skin, and injected recombinant murine interferon gamma. Alopecia areata is a complex polygenic trait with low penetrance in these mice. Injection of interferon gamma did not change the frequency or time of onset of alopecia in these mice suggesting this protein alone is not sufficient to initiate disease.     

A point mutation in the tyrosine hydroxylase gene associated with Segawa's syndrome

We have examined the molecular basis of Segawa's syndrome in six families with seven affected children. In one family two siblings with this disease carried a point mutation in exon 11 of the tyrosine hydroxylase gene, resulting in an amino acid exchange of Gln³⁸¹ to Lys³⁸¹. These results suggest that a change in tyrosine hydroxylase causes this form of Segawa's syndrome.     

Isoosmotic isolation of rat brain synaptic vesicles, some of which contain tyrosine hydroxylase

Rat brain synaptic vesicles were isoosmotically isolated and examined for Mg(2+)-ATPase [EC 3.6.1.3.] and tyrosine hydroxylase [EC 1.14.16.2.] associated with the synaptic vesicles. Synaptosomes in 0.32 M sucrose were disrupted by freezing and thawing treatment, and the cytosol fraction was fractionated on a Sephacryl S-500 column with a mean exclusion size of 200 nm. Peak I at the void volume was a mixture of large vesicular membranes, small amounts of synaptic vesicles and coated vesicles, etc. Peak II consisted of non- and granulated synaptic vesicles of 35-40 nm diameter, and peak III of soluble proteins. The synaptic vesicles in peak II reacted with antibodies against the H(+)-ATPase A-subunit, vesicular acetylcholine transporter, and vesicular monoamine transporter. However, they showed little Mg(2+)-ATPase activity. Tyrosine hydroxylase was observed in either peak II or III on blotting with an anti-tyrosine hydroxylase antibody. These results imply that tyrosine hydroxylase exists in soluble and bound forms to synaptic vesicles in nerve terminals.