Interleukin-2 receptor alpha-chain expression in patients with alopecia areata

Interleukin-2 (IL-2) is a lymphokine produced by activated T-cells. Its receptor, IL-2R, is expressed on T-cells. Several clinical and experimental findings point towards IL-2 as a crucial mediator inducing immunologic reaction against human follicle in alopecia areata. The objective of our study was to analyze the expression of IL-2R as a sign of T-cell activation in scalp biopsies of patients suffering from alopecia areata. An immunohistochemical analysis was used to determine the difference in cytokine-regulated expression of IL-2R between 45 patients with active and stable phase of alopecia areata and 23 healthy control subjects. In the patients with alopecia areata in active phase, the expression of IL-2R in scalp biopsies was significantly stronger than that in the patients with stable disease and in controls. The increase of IL-2R+ cells in early phase of the disease could suggest that T-lymphocyte activation with IL-2 secretion and IL-2R expression may initiate the immune inflammatory mechanism of alopecia areata.     

The TRAF1/C5 locus confers risk for familial and severe alopecia areata

Background Alopecia areata (AA) is a common hair loss disorder with a complex mode of inheritance. Autoimmune mechanisms are presumed to be crucial aetiologically. It is plausible that a number of autoimmune disorders may share a common genetic background. This phenomenon has been demonstrated in previous studies, which have shown an overlap of susceptibility alleles between AA and other autoimmune disorders. Recent studies have shown that genetic variants on the TRAF1/C5 (tumor necrosis factor receptor‐associated factor 1, complement component 5) locus confer susceptibility to rheumatoid arthritis (RA). Objectives To examine the role of the TRAF1/C5 locus in the development of AA using a large sample of 1195 patients with AA and 1280 controls. Methods We genotyped the two most significant single nucleotide polymorphisms (SNPs) (rs10818488, rs2416808) from a former RA candidate gene study. After having obtained evidence for association, we performed a fine‐mapping study and genotyped the locus with an additional 27 SNPs. Results While no significant result was obtained for the overall sample, rs2416808 showed significant associations in the analysis of the subgroups with severe AA and with a positive family history. The most significant P‐value for rs2416808 was in familial cases (P = 0·004, P_corr = 0·026). The fine mapping revealed significant associations for four additional SNPs in the analysis of subgroups, with rs2416808 remaining the most significant marker. Conclusions Our results point to the involvement of the TRAF1/C5 locus in the aetiology of familial and severe AA, and provide further support for a shared aetiology between AA and other autoimmune disorders.     

Interleukin-1 receptor antagonist production by human keratinocytes.

Human keratinocytes produce biologically active pro-IL-1 alpha and inactive pro-IL-1 beta with most protein remaining intracellular. IL-1 receptor antagonist (IL-1ra) is a newly described member of the IL-1 family that is secreted by stimulated monocytes and binds competitively to IL-1 receptors without stimulating target cells. We examined the characteristics of IL-1ra production by cultured human keratinocytes. By ELISA, keratinocyte lysates contained 390 ng IL-1ra/mg total protein with little IL-1ra detected in supernatants. In contrast, monocytes produced 297 ng IL-1ra/mg total protein during 24 h of culture on adherent IgG with about half of the IL-1ra detected in supernatants. By Western blot analysis, keratinocyte IL-1ra was approximately 20 kD in size and was slightly larger than recombinant monocyte IL-1ra. In contrast to monocytes, human keratinocyte IL-1ra was not secreted in 22-25-kD molecular weight glycosylated forms. Affinity-purified keratinocyte IL-1ra exhibited identical biologic activity to recombinant monocyte IL-1ra, each inhibiting IL-1-dependent augmentation of murine thymocyte proliferation to the same degree per amount of protein. An IL-1ra mRNA of 1.8 kb was detected by Northern blot analysis in RNA extracted from keratinocytes. In order to determine the effect of differentiation on IL-1 and IL-1ra production, human keratinocytes were cultured for 72 h in low (0.03 mM), medium (0.15 mM), or high (1.0 mM)-calcium concentrations. The absolute amounts of IL-1ra increased twofold and the ratio of IL-1ra to IL-1 alpha in keratinocyte lysates increased from approximately 12:1 to 25:1 during differentiation. These results indicate that keratinocytes constitutively produce large amounts of a biologically active intracellular variant of IL-1ra that increase with differentiation. IL-1ra released during keratinocyte damage may be important in modifying the inflammatory effects of IL-1 alpha in human skin.     

The genetic basis of alopecia areata: HLA associations with patchy alopecia areata versus alopecia totalis and alopecia universalis

Many diseases, notably those having a strong autoimmune component, have been shown to have an association with specific human leukocyte antigens (HLA). The molecular basis for this genetic association with disease is the fact that HLA bind and present peptides derived from self and foreign protein antigens to the immune system for recognition and activation of the immune response. Previous studies with heterogeneous groups of alopecia areata (AA) patients have suggested associations with some HLA class I and class II antigens. For this study we selected only patients with long-standing disease and stratified them into two groups by strict definitions of duration and extent of disease: those with patchy AA and those with either alopecia totalis (AT) or alopecia universalis (AU). The patients were tissue typed for HLA class II antigens by biomolecular methods that provided antigen discrimination at an allele level. More than 80% of all of the AA patients typed were positive for the antigen DQB1*03 (DQ3), suggesting that this antigen is a marker for general susceptibility to AA. In addition, two other antigens were found significantly increased in frequency only in the group of AT/AU patients, DRB1*0401 (DR4) and DQB1*0301(DQ7). This strongly suggests that the two clinical types of AA, namely patchy AA versus AT/AU, can be distinguished by a genetically based predisposition to extent of disease.     

Melanocortin receptor type 2 (MC2R,ACTH receptor) expression in patients with alopecia areata

Please cite this paper as: Melanocortin receptor type 2 (MC2R, ACTH receptor) expression in patients with alopecia areata. Experimental Dermatology 2010; 19 : 1020–1022. Abstract: Human skin expresses elements of the hypothalamo‐pituitary‐adrenal (HPA) axis that function as a local stress response system. Because adrenocorticotropic hormone (ACTH) is an intermediate in the HPA axis from corticotropin‐releasing hormone (CRH) signal to cortisol secretion, MC2R that binds only ACTH may be important in the stress response of skin. We investigated the local expression of MC2R by immunohistochemistry to identify the role of ACTH/MC2R in stress‐associated alopecia areata (AA). MC2R appeared to be highly compartmentalized in scalp skin including the epidermal cells of hair follicles and epidermis, sebaceous and eccrine glands, as well as dermal fibroblasts. The expression of MC2R was lower in AA lesions than in normal scalp tissue in almost all scalp skin cells, especially in epithelial cells. These findings demonstrate that MC2R expression is aberrant in AA and suggest a deficit in ACTH/MC2R activity may play an important role in the pathophysiology of AA.     

CD44 expression in alopecia areata and androgenetic alopecia

CD44 is a widely distributed cell surface protein thought to be involved in multiple steps of normal immune cell function, including T-cell activation, and in cellular adhesion where it mediates cell attachment to hyaluronate. In normal skin, CD44 is found by immunohistochemical means to be primarily in eccrine coil cells. In this study, we have looked at the expression of CD44 in normal scalp and in two different hair disorders, androgenetic alopecia and alopecia areata. In normal scalp and androgenetic alopecia, CD44 was found in its normal distribution in eccrine coil cells. In scalp of 30 patients with alopecia areata, there was no expression of this glycoprotein. Patients were also assessed pre and post treatment for their alopecia areata, and even though they had no significant hair regrowth, 2 patients regained expression of CD44, indicating a variable expression of this protein in the alopecia areata disease process. The absence of CD44 expression in alopecia areata-affected scalp may give further information regarding the pathogenesis of this disease.     

Lack of association between Vitamin D receptor FokI polymorphism and alopecia areata

Vitamin D receptor (VDR) is expressed in the hair follicle and the lack of it leads to alopecia. In this study, we investigated whether there was a relationship between VDR FokI gene polymorphism and alopecia areata (AA). This is the first study investigating the relationship between VDR gene polymorphism and AA. Twenty-five patients with the extensive forms of AA (alopecia totalis; AT, alopecia universalis; AU and AT/AU) and 27 healthy control subjects were genotyped. Their genotypes were determined by a polymerase chain reaction (PCR) and restriction fragment length polymorphism analysis. The genotypes were classified as FF (absence of the FokI site) and ff (presence of the FokI site). Allele frequencies for F and f alleles were 76.0% and 24.0% in the alopecic group and 72.2% and 27.7% in the control group (p > 0.05). The frequencies for the FF, Ff and ff genotypes were 56.0%, 40.0% and 4.0% in the patient group, and 48.1%, 48.1% and 3.7% in the control group, respectively. No statistically significant differences were observed in the frequencies of the VDR FokI genotype between the patient and the control groups. However, to conclude that there is no relationship between VDR gene polymorphism and AA, the VDR FokI polymorphism should be further studied in other populations, larger groups, and the distribution of other VDR polymorphisms such as BsmI, Tru9I, ApaI, TaqI and polyA.     

Vitamin D receptor gene polymorphisms are not associated with alopecia areata

BACKGROUND: It has been demonstrated that the vitamin D receptor (VDR) is strongly expressed in key structures of hair follicles, and a lack of VDR leads to alopecia. We investigated whether there was any association between VDR gene polymorphisms (BsmI, ApaI, and TaqI) and alopecia areata (AA). METHODS: Thirty-two patients with AA and 27 healthy control subjects were genotyped using polymerase chain reaction and restriction fragment length polymorphism analysis. RESULTS: In the patient group, the B and b allele frequencies were 53.1% and 46.9%, A and a allele frequencies were 70.3% and 29.7%, and T and t allele frequencies were 62.5% and 37.5%, respectively. In the control group, the corresponding values were 51.9% and 48.1%, 63.0% and 37.0%, and 77.8% and 22.2%, respectively. In the patient group, the BB, Bb, and bb genotype frequencies were 25.0%, 56.2%, and 18.8%, AA, Aa, and aa genotype frequencies were 43.8%, 53.1%, and 3.1%, and TT, Tt, and tt genotype frequencies were 37.5%, 50.0%, and 12.5%, respectively. In the control group, the corresponding values were 11.1%, 81.5%, and 7.4%, 29.6%, 66.7%, and 3.7%, and 63.0%, 29.6%, and 7.4%, respectively. None of the allele or genotype frequencies showed statistically significant differences between the patient and control groups. CONCLUSION: These findings suggest that there is no relationship between VDR gene polymorphisms and AA.     

Investigation of the HLA-DRB1 locus in alopecia areata

To further evaluate the nature of the HLA association with alopecia areata (AA), we investigated the HLA-DRB1 locus in 161 AA patients and 165 matched controls from Belgium and Germany. HLA-DRB1 typing was performed using a recently established method that employs a combination of PCR-SSP (sequence specific priming) and Pyrosequencing(TM) technology. No significant differences were observed for HLA allele groups DRB1 *01, *07, *08, *09, *10, *11, *13, *14, *15, and *16. HLA-DRB1*03 was found to confer a protective effect (7.5% versus 13.6%, p = 0.011). Additional genotyping at the allelic level revealed a significant difference in HLA-DRB1*0301 between patients and controls (6.8% versus 11.2%, p = 0.048). The DRB1*04 allele group was confirmed as a risk factor for the development of AA (20.8% versus 13.3%, p = 0.012), with the allele DRB1*0401 accounting for the greatest proportion of the effect (13.4% versus 7.3%, p = 0.014). Results obtained after subgrouping of the patients according to age at onset, severity and family history of the disease suggests that the genetic effects of the HLA system are strongest in familial cases of the disease.     

Oral cyclosporin and alopecia areata.

A 30-year-old woman with severe alopecia areata of the scalp was treated with oral cyclosporin for 3 months. Clinical, histological and immunohistochemical assessments failed to reveal any improvement.     

Epidemiology and genetics of alopecia areata

The frequency of alopecia areata and observed patterns of heritability are in keeping with a polygenic inheritance model but the genetics of alopecia areata is still poorly understood. The role of environmental factors in triggering disease initiation or exacerbation remains almost entirely speculative. Using the candidate gene approach, three susceptibility/severity factors have been identified. HLA alleles were the first to show a strong association with alopecia areata and some DQB and DR alleles have been demonstrated to confer a high risk for disease by both case-control and family-based studies. Interleukin (IL)-1 cluster genes, mainly the IL-1 receptor antagonist, show a strong association with disease severity in alopecia areata and a number of other autoimmune and inflammatory diseases. Finally, the association of alopecia areata with Down's syndrome, the high frequency of alopecia areata in autoimmune polyglandular syndrome type I due to mutations of the autoimmune regulator (AIRE) gene on chromosome 21q22.3 and the finding of association with MX1, another gene in the Down's syndrome region of chromosome 21 indicate this area of the genome as a promising target for future-family based investigations. The role of individual genes of the MHC, IL-1 cluster or chromosome 21q22.3 is difficult to establish and further genetic and functional investigations are needed to confirm their involvement in the pathogenesis of alopecia areata.     

Colocalization of alopecia areata and vitiligo

We report an 18-year-old woman who had a 4-month history of colocalization of alopecia areata and vitiligo, principally on the occipital portion of the scalp.     

Diagnosis and therapy of alopecia areata

In general, the diagnosis of alopecia areata is not difficult. In children, we have to consider trichotillomania as a differential diagnosis; in adult patients pseudopelade and syphilis have to be taken into account. Investigation of the nails may support the diagnosis and help to assess the degree of activity of the disease. The efficacy of topical immunotherapy using diphencyprone or squaric acid dibutylester has been proved by various study groups. Unilateral comparative studies supported our assumption that the therapeutic response is most probably due to a local immunomodulation. The applicability of this method is limited by the fact that the toxicological data available on the contact allergens used are still incomplete.