Journal of Investigative Dermatology

Published by Elsevier BV

Online ISSN: 1523-1747


Print ISSN: 0022-202X


The Environment and Human Health
  • Article

June 2006


52 Reads

David A Schwartz

Circadian Activity of Topical 0.05% Betamethasone Dipropionate in Human Skin In Vivo

June 1994


82 Reads

The influence of treatment duration, vehicle, and time of day of application on topical 0.05% betamethasone dipropionate uptake into human stratum corneum and the resulting skin-blanching response was investigated in human subjects. Drug uptake into stratum corneum and the resulting skin color changes measured with a chromameter demonstrate an equilibrium delay. Maximal drug uptake occurred at 2 h, whereas maximal skin color changes occurred 6 h after a single application. Extent of decreased skin color was dependent on vehicle, treatment duration, and time of day of application. Time of maximal decreased skin color occurred at midnight independent of vehicle, treatment duration, or time of day of application. This time of maximal drug activity coincides with the well-known time period of lowest circulating cortisol concentrations (2000-0400 h). Application of a single 2- or 6-h dose of the 0.05% cream at 1600 h produced more extensive and prolonged changes in skin color over 24 h than a 0900-h application in the same subject. These data demonstrate that the extent and duration of topical corticosteroid activity in human skin is influenced by vehicle, treatment duration, and time of day of application. The prolonged changes in skin color measured with a single dose applied at 1600 h suggest that a once-a-day dosing regimen in the late afternoon may be sufficient for dermatologic therapy. Elucidation of these circadian responses with topical corticosteroids may provide a rational basis for the future re-evaluation of the appropriate therapeutic regimen with this class of drugs in dermatologic medicine.

Pharmacokinetics of 0.1% Tacrolimus Ointment After First and Repeated Application to Adults with Moderate to Severe Atopic Dermatitis

July 2005


130 Reads


Rita Gutmane


Nora Valdmane




The systemic exposure to tacrolimus after first and repeated application of 0.1% tacrolimus ointment was investigated in 32 adults with moderate to severe atopic dermatitis. Patients were allocated to treatment groups according to the size of the affected area to be treated: Group 1</=3000 cm(2) (N=11); Group 2>3000 cm(2)</=6000 cm(2) (N=12); Group 3>6000 cm(2)</=10,000 cm(2) (N=9). Ointment was applied twice daily for 13 d and once daily on Day 14; the size of application area remained the same irrespective of healing. Blood samples were collected on Days 1 (first application), 4, and 14 (last application) and analyzed by a validated HPLC-MS/MS method. Systemic exposure to tacrolimus was generally low with 96% of blood samples assayed containing concentrations below 1 ng per mL and 23% of samples below the lower limit of quantification (0.025 ng per mL). Peak concentrations after first ointment application were </=2.8 ng per mL, and the mean area under the concentration-time curve between 0 and 12 h using the trapezoidal rule (AUC(0-12)) values were 1.1, 1.6, and 4.8 ng h per mL for Groups 1, 2, and 3, respectively. The corresponding mean values on Day 14 were similar indicating negligible systemic accumulation of tacrolimus after repeated ointment applications. Both the rate and extent of topical absorption decreased as the skin lesions healed.

Studies by electron microscopy on epidermis after topical application of mercuric chloride: Morphologic and histochemical findings in epidermal cells of human subjects who do not show allergic or primary intact reaction to mercuric chloride (0.1%)

March 1971


24 Reads

The morphologic and histochemical changes seen by electron microscopy after a single topical application of mercuric chloride 10.1% to the skin of subjects not sensitive to it were studied. Whereas signs of injury are not detectable clinically in subjects not sensitive to mercury, they can be detected at the ultrastructural level. The ultrastructure shows varying degrees of cell degeneration which becomes more pronounced the longer the exposure to mercuric chloride. After topical application of mercuric chloride the following was observed: 1) glycogen deposits appeared in cytoplasm of some Langerhans' cells: 2) lysosome-like bodies appeared in keratinocytes, Langerhans' cells and rarely in melanocytes; 3) electron-dense deposits were demonstrable in keratinocytes, Langerhans' cells and melanocytes after specimens were processed with glutaraldehyde, ammonium sulfide and osmium tetroxide; and 4) cells with features of both kerationcytes and Langerhans' cells appeared. Cells in different parts of each specimen showed various degrees of involvement. The findings reported here serve as a basis for comparison of changes seen in clinically reactive skin sites in allergic sensitivity or primary irritant reactions to mercuric chloride.

Figure 1: Flow diagram of the study.Download Power Point slide (238 KB)
Maintenance Therapy of Adult Vitiligo With 0.1% Tacrolimus Ointment: A Randomized, Double Blind, Placebo-Controlled Study.
  • Article
  • Full-text available

December 2014


399 Reads

The risk of relapse after successful repigmentation in vitiligo is estimated to 40% within the first year. It has been shown in atopic dermatitis that continuous low-level use of topical corticosteroids and calcineurin inhibitors in previously affected skin can prevent new flares.We hypothesized that a twice-weekly application of 0.1% tacrolimus ointment might be effective for maintaining repigmentation in therapeutically repigmented lesions of vitiligo patients. After randomization, sixteen patients with 31 patches were assigned to the placebo group and 19 patients with 41 patches were assigned to the tacrolimus group. In the intention to treat analysis, 48.4% of lesions showed depigmentation in the placebo group whereas 26.8% did in the tacrolimus group (p=0.059). The intention to treat results did not remain significant after adjustment for within-patient clustering, OR 2.55; 95% CI [0.65-9.97] p=0.1765). The per protocol analysis (n=56), showed that 40% of lesions had some depigmentation in placebo group whereas only 9.7% did in tacrolimus group (p=0.0075). The per protocol results remained significant after adjustment for within-patient clustering: OR 6.22; CI 95% [1.48-26.12] p=0.0299. Our study shows that twice-weekly application of 0.1% tacrolimus ointment is effective in preventing the depigmentation of vitiligo patches that have been previously successfully repigmented.Journal of Investigative Dermatology accepted article preview online, 18 December 2014. doi:10.1038/jid.2014.527.

Figure 1: Inhibition of intracellular signaling in keloid fibroblast (KF) by both KU-0063794 and KU-0068650. (a) Differential expression of mammalian target of rapamycin (mTOR) and phosphor-mTOR (p-mTOR) (n=6). (b) Differential expression of mTOR signaling in KFs and extra-lesional fibroblasts (ELFs) using In-Cell Western Blotting (ICWB) (n=11). (c) ICWB average immunoreactivity from (b). (d) ICWB average immunoreactivity of KF (n=11) induced mTOR inhibitors, normalized to β-actin. (e) Both KU-0063794 and KU-0068650 inhibit mTORC1 and mTORC2 in primary KFs. Bar graphs represent the quantification of average protein expression in different treatments from three independent ICWB experiments (n=6). (f) Co-immunoprecipitation for mTOR with raptor and Rictor to assess TORC1 and TORC2 complex inhibition by both the compounds (n=11). The data presented here are the means±SEM of triplicate experiments performed. **P<0.05, *P0.01 indicate significant difference in the treated group compared with the DMSO control group. AKT, also known as protein kinase B (PKB); 4E-BP1, eukaryotic initiation factor 4 E-binding protein-1; GSK3β, glycogen synthase kinase-3; HIF-1α, hypoxia-inducible factor-1alpha; IP, immunoprecipitation; MAPK, mitogen-activated protein kinase; p-MAPK, phosphorylated mitogen-activated protein kinase; Raptor, regulatory associated protein of mTOR; Rictor, rapamycin-insensitive companion of mTOR.Download Power Point slide (512 KB)
Figure 2: Role of mammalian target of rapamycin (mTOR) inhibitors on keloid cell viability/metabolic activity by water-soluble tetrazolium salt-1 (WST-1), cell adhesion, spreading, and proliferation by real-time cell analysis (RTCA) on microelectronic sensor arrays. (a) WST-1 assay was performed 24 hours post treatment with different mTOR inhibitors, for viability/metabolic activity (keloid fibroblast (KF): n=8). (b) WST-1 assay for extra-lesional fibroblasts (n=5). (c) Quantitative analysis of RTCA average cell index of KFs (n=9). Primary KFs were seeded on 96-well E-plate (1 × 104 cells per well) and cells were treated with different mTOR inhibitor concentrations. Cell index (CI) on RTCA was recorded every 15 minutes. **P<0.04, *P0.01 indicate significant difference compared with the DMSO control group. The data expressed are an average means±SEM from four independent experiments.Download Power Point slide (368 KB)
Figure 3: Differential effect of KU-0063794, KU-0068650, and Rapamycin on primary keloid fibroblast (KF) and extra-lesional fibroblast (ELF) migration and invasion properties. (a) Fibroblast migration response toward the 2-mm migration zone, in an in vitro two-dimensional collagen assay. Representative micrographs are shown from three independent experiments. (b) The average number of migrated fibroblasts in the migration zone with and without compounds. *P<0.05, significant difference compared with the DMSO group. **P0.01, significant difference compared with the Rapamycin group. #P<0.05, significant difference in KF migration compared with ELFs. (c) Fibroblasts’ invasive response toward the 2-mm invasion zone in an in vitro three-dimensional basement membrane extract model. (d) The average number of invaded cells in the invasion zone. **P<0.01 indicates significant difference compared with primary ELFs. *P<0.03 indicates significant difference in primary KFs compared with the DMSO group. #P<0.05 indicates significant difference in primary ELFs compared with the DMSO group.Download Power Point slide (524 KB)
Figure 4: Effect of KU-0063794 and KU-0068650 compounds compared to Rapamycin in in vitro and ex vivo keloid models. (a) Both KU-0063794 and KU-0068650 inhibit the expression of collagen, fibronectin, and α-smooth muscle actin (α-SMA) at messenger RNA (mRNA) levels (keloid fibroblast (KF): n=8). (b) In-Cell Western Blotting analysis of the expression of ECM proteins, Cyclin D, and proliferating cell nuclear antigen (PCNA), 24 hours post treatment with different mammalian target of rapamycin inhibitors (KF: n=8). (c) Shrinkage of keloid organ culture (OC) after different compound treatments. Average weight of the keloid OC (n=10) at different time points are indicated in the bar graph. Four millimeter keloid explant biopsies were removed from the collagen gel matrix and its weight was determined in triplicates. (d) In MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay both, KU-0063794 and KU-0068650 showed much greater inhibitory effect on metabolic activity in keloid OC (n=8) as compared with Rapamycin. *P<0.05, **P0.01 indicates significant difference compared with the DMSO group.Download Power Point slide (424 KB)
Figure 5: Both KU-0063794 and KU-0068650 compounds induce apoptosis and deplete CD31 and CD34 +Ve cells in keloid organ culture. (a) Representative micrographs of TUNEL staining (red-nuclei and green–yellow TUNEL+Ve cells) (n=8). Original magnification, × 200. D, dermis; EP, epidermis. Arrows indicate TUNEL+Ve cells. (b) % of TUNEL+Ve cells (n=8). *P<0.05, **P<0.01 indicates significant difference compared with the DMSO control group. (c) Representative micrographs of CD31+Ve endothelial cells after different compound treatments (n=8). Original magnification × 100. Arrows indicate CD31+Ve cells. (d) Representative micrographs of CD34+Ve microvascular endothelial cell marker after different compound treatments. Original magnification, × 200. Red, nuclei; green, CD34+Ve cells. Arrows indicate CD34+Ve cells.Download Power Point slide (582 KB)
Potent dual inhibitors of TORC1 and TORC2 complexes (KU-0063794 and KU-0068650) demonstrate in vitro and ex vivo anti-keloid scar activity

January 2013


216 Reads

Mammalian target of rapamycin (mTOR) is essential in controlling several cellular functions. This pathway is dysregulated in keloid disease (KD). KD is a common fibroproliferative dermal lesion with an ill-defined treatment strategy. KD demonstrates excessive matrix deposition, angiogenesis, and inflammatory cell infiltration. In KD, both total and phosphorylated forms of mTOR and p70(S6K)(Thr421/Ser424) are upregulated. Therefore, the aim of this study was to investigate adenosine triphosphate-competitive inhibitors of mTOR kinase previously unreported in keloid and their comparative efficacy with Rapamycin. Here, we present two mTOR kinase inhibitors, KU-0063794 and KU-0068650, that target both mTORC1 and mTORC2 signaling. Treatment with either KU-0063794 or KU-0068650 resulted in complete suppression of Akt, mTORC1, and mTORC2, and inhibition of keloid cell spreading, proliferation, migration, and invasive properties at a very low concentration (2.5 μmol l(-1)). Both KU-0063794 and KU-0068650 significantly (P<0.05) inhibited cell cycle regulation and HIF1-α expression compared with that achieved with Rapamycin alone. In addition, both compounds induced shrinkage and growth arrest in KD, associated with the inhibition of angiogenesis, induction of apoptosis, and reduction in keloid phenotype-associated markers. In contrast, Rapamycin induced minimal antitumor activity. In conclusion, potent dual mTORC1 and mTORC2 inhibitors display therapeutic potential for the treatment of KD.Journal of Investigative Dermatology advance online publication, 10 January 2013; doi:10.1038/jid.2012.483.

The Association Between HLA DR, DQ Antigens, and Vulval Lichen Sclerosus in the UK: HLA DRB1&ast;12 and its Associated DRB1&ast;12&sol;DQB1&ast;0301&sol;04&sol;09&sol;010 Haplotype Confers Susceptibility to Vulval Lichen Sclerosus, and HLA DRB1&ast;0301&sol;04 and its Associated DRB1&ast;0301&sol;04&sol;DQB1&ast;0201&sol;02&sol;03 Haplotype Protects from Vulval Lichen Sclerosus

November 2005


137 Reads

Lichen sclerosus (LS) is considered to have an immunogenetic background. Several small studies, using serological typing, have reported that HLA-DR11, DR12, and DQ7 were increased in LS, with DR17 less frequent. This study aimed to validate and detect new HLA-DR and DQ associations with LS in females and its characteristic clinical parameters. The cases, 187 female LS patients, and 354 healthy controls were all UK North Europeans. PCR-sequence specific primers method was applied to genotype the HLA-DR, DQ polymorphisms that correspond to 17 serologically defined DR and seven DQ antigens. Statistical analysis was performed with two-tailed Fisher's exact test with Bonferroni adjustment (p value after Bonferrroni adjustment, Pc). We found increased frequency of DRB1*12 (DR12) (11.2%vs 2.5%, pc < 0.01) and the haplotype DRB1*12/DQB1*0301/04/09/010 (11.2%vs 2.5%, p < 0.001, pc < 0.05), and a lower frequency of DRB1*0301/04 (DR17) (11.8%vs 25.8%, pc < 0.01) and the haplotype DRB1*03/DQB1*02DRB1*0301/DQB1*0201/02/03 (11.2%vs 24.6%, pc < 0.0001) in patients compared with controls. HLA DR and DQ antigens were not associated with time of onset of disease, site of involvement, structural changes of genitals, and response to treatment with potent topical steroids. In conclusion, HLA-DR and DQ antigens or their haplotypes appear to be involved in both susceptibility to and protection from LS.

The Role of Active Oxygen (102 and 02-) Induced by Crude Coal Tar and Its Ingredients Used in Photochemotheraphy of Skin Diseases

February 1984


31 Reads

Crude coal tar (CCT) and certain photoreactive ingredients of CCT are photosensitizing agents used in the treatment of skin diseases (psoriasis, atopic eczema, etc.). Limited information is available in elucidating the mode of action of CCT in clearing psoriasis or causing skin photosensitization reactions. The production of singlet oxygen (1O2) and superoxide radicals (O(2) or HO2), the formation of interstrand cross-links (ICL) in DNA, and the skin photosensitization reaction caused by CCT or the ingredients present in tar preparations have been examined. Both type I (oxygen-independent) and type II (sensitized reactions requiring molecular oxygen) reactions are induced by CCT. Our data show that CCT and some of the photoreactive ingredients present in CCT produce 1O2, O(2), and ICL in DNA upon exposure to UVA radiation. Based on the equivalent concentration, the efficiency of various agents to produce 1O2 was of the following order: hematoporphyrin greater than phenanthridine greater than acridine greater than methylene blue greater than CCT greater than fluoranthrene greater than anthracene greater than pyrene greater than 8-methoxypsoralen greater than anthralin greater than chloroquine greater than anthralin dimer. The O(2) formation with CCT and its ingredients was also of the same order except for anthracene which was found to be a strong producer of O(2). The therapeutic effectiveness of CCT appears to be due to: (a) its cytotoxic effects, and (b) the production of 1O2, O(2), and ICL by CCT and its photoreactive ingredients. The skin photosensitizing (smarting, edema, and erythema responses) and carcinogenic properties of CCT may also be related to the production of 1O2 and O(2) and the formation of ICL which appear to be responsible for inducing the damage to the DNA and cell membrane.

Figure 1. EH-57.1-positive individuals carry a 26 times elevated risk of developing type I psoriasis. HLA class I antigens were analyzed by standard serological methods, and HLA class II alleles were typed by PCR-SSO. Absolute numbers of cases positive for the allele or alleles under investigation were determined and used to calculate phenotype frequencies as well as 2 likelihood ratios. The p-values were corrected according to Bonferroni. Phenotype-frequencies (Pf) of the single HLA class I and II alleles within EH-57.1 and of EH-57.1 itself are shown. Controls (open columns) are compared with type I and type II psoriasis (solid columns). Significantly overrepresented (p cor. 0.01) and highly significantly overrepresented alleles (p cor. 0.001) are denoted by * and **, respectively. RR values were computed according to Woolf (1955) and are demonstrated by black diamonds. 
Figure 2. The HLA class I, but not the class II side of EH-57.1, confers susceptibility to type I psoriasis. Phenotype frequency (Pf) of haplotypes containing the class I antigens of EH-57.1 (Cw6-B57) but lacking the class II alleles of this EH (DRB1*0701-DQA1*0201DQB1*0303) (referred to as the class I side) and of haplotypes positive for the class II alleles of EH-57.1 in the absence of its HLA-class I alleles (referred to as the class II side) in controls (open columns) and type I psoriasis (solid columns). 
Familial Juvenile Onset Psoriasis Is Associated with the Human Leukocyte Antigen (HLA) Class I Side of the Extended Haplotype Cw6-B57-DRB1*0701-DQA1*0201-DQB1*0303: A Population- and Family-Based Study

May 1996


115 Reads

To further evaluate the nature of the HLA association with psoriasis, HLA haplotypes of 60 patients with type 1 (early onset, positive family history) and 30 patients with type II (late onset, no family history) psoriasis were investigated by polymerase chain reaction sequence-specific oligonucleotide hybridization (HLA class II) and serology (HLA class I). Ethnically matched blood donors (146) served as controls. In type I, but not type II psoriasis, the Caucasian HLA extended haplotype (EH) Cw6-B57-DRB1*0701-DQA1*0201-DQB1*0303 named according to the B allele EH-57.1 was highly significantly overrepresented (p cor= 0.00021). This particular EH was present in 35% of type I psoriatics but only 2% of controls. EH-57.1+ individuals therefore carry a 26 times higher risk of developing type I psoriasis than individuals who are EH-57.1-negative Further analysis of individual HLA alleles revealed that within EH-57.1, HLA class I antigens (Cw6-B57) were associated to a much higher extent with type I psoriasis than the HLA class II alleles (DRB1*0701-DQA1*0201-DQB1* 0303). Pedigree analysis of three multiply affected families over three generations revealed a cosegregation of disease with EH-57.1. These results strongly suggest that a gene for familial psoriasis is associated with the class I side of the extended haplotype Cw6-B57-DRB1*0701-DQA1*0201-DQB1*0303.

Oligonucleotide typing reveals association of type I psoriasis with the HLA-DRB1*0701/2, -DQA1*0201, -DQB1*0303 extended haplotype

July 1993


112 Reads

Although the pathogenesis of psoriasis is still a matter of debate, there are several lines of evidence supporting the concept of this disease being immunologically mediated with T cells playing a crucial role. Because a considerable portion of the cellular infiltrate in psoriasis consists of activated T-helper cells, expression of HLA class II antigens might be of particular importance for the understanding of its pathogenesis. Therefore, we investigated the HLA type of patients with type I (early onset, positive family history) and type II (late onset, no family history) psoriasis by means of serology (n = 89) and genotyping using sequence-specific oligonucleotide probes (n = 64). Serologic analysis of class I documented the association of type I psoriasis with HLA-Cw6, -B13, and -B57, whereas type II psoriasis showed a weaker correlation with HLA-Cw2 and -B27. Genotyping using SSO for class II detected the elevation of the HLA-DRB1*0701/2 allele frequency from 13% in normal population to 36% in type I, but only to 15% in type II psoriatics. Moreover, positive correlations with type I psoriasis were detected for HLA-DQA1*0201 and HLA-DQB1*0303. The HLA-DRB1*0701/2, -DQA1*0201, -DQB1*0303 extended haplotype was found exclusively in type I psoriasis. This is the first report documenting the association of distinct HLA class II alleles with type I psoriasis as detected on the DNA level, an approach both more specific and more sensitive when compared to serology.

HLA-A*0201+ Plasmacytoid Dendritic Cells Provide a Cell-Based Immunotherapy for Melanoma Patients

June 2012


64 Reads

Several sources of evidence suggest that tumor-specific T cells have the potential to control melanoma tumors. Current active and adoptive therapeutic approaches to elicit such T cells are either not sufficiently clinically efficient or require fastidious processes that impede their extensive clinical use. As plasmacytoid dendritic cells (pDCs) have a crucial role in triggering antitumor immunity especially in melanoma, we explored their potential as a cell-based approach for melanoma immunotherapy. An irradiated human HLA-A(*)0201(+) pDC line loaded with peptides derived from the major melanoma tumor antigens, MelA/MART-1, gp100/pmel17, tyrosinase, and MAGE-A3, was used to trigger functional multi-specific T cells ex vivo from peripheral blood mononuclear cells and tumor-infiltrating lymphocytes from stage I-IV HLA-A(*)0201(+) melanoma patients. pDCs loaded with melanoma-derived peptides promptly induced high levels of melanoma tumor-specific T cells from both sources. pDC-primed central/effector memory antitumor T cells were highly functional as indicated by the specific IFNγ secretion and membrane CD107 expression upon stimulation. Cells also exhibited strong cytotoxicity toward semi-allogeneic melanoma cells and patient-derived tumor cells. The simple design and potent efficacy of this promising approach provides a preclinical basis for the development of a pDC-based vaccine and an alternative means to produce tumor-specific T cells for adoptive cellular immunotherapy in melanoma patients.

Welsh EA, Clark HH, Epstein SZ, Reveille JD, Duvic MHuman leukocyte antigen-DQB1*03 alleles are associated with alopecia areata. J Invest Dermatol 103:758-763

January 1995


27 Reads

Alopecia areata (AA) is characterized by hair loss in patches (patchy AA), over the entire scalp (AT, totalis), or universally (AU). An autoimmune mechanism has been hypothesized, because the inflammatory infiltrate targeted to the hair follicles includes activated T cells. To investigate whether or not genetic polymorphism of the human leukocyte antigen (HLA) region contributes to disease susceptibility, we used sequence-specific oligonucleotides and amplified genomic DNA to define HLA-DQA1, -DQB1, and -DPB1 alleles in a cohort of 85 white patients. The frequency of DQB1*0301 was significantly increased to 41% in all patients, and to 47% in AT/AU patients relative to controls (27%). Analyzed together, DQB1*03 alleles (DQB1*0301-*0303) were increased to 80% (all patients) and to 92% (AT/AU) (odds ratio = 12.14, p = 0.00003, corrected). This striking association implicates the DQB1*03 alleles in the pathogenesis of AA. DQB1*06 was decreased relative to controls (56%) in all patients (32%, odds ratio = 0.37, p = 0.0045, corrected). An increase was observed in the HLA-DRB1*11(DR5) allele DRB1*1104, which may result from linkage disequilibrium with DQB1 alleles. Sequence comparison among the allele products associated with AA indicates that the DQB1*03 alleles carry a unique proline at position 55 that is not present in alleles that are neutral or negatively associated with the disease. This highly significant association may exert considerable control over immune responsiveness and the initiation or persistence of a T-cell autoimmune response against the hair follicle.

Black Patients of African Descent and HLA-DRB1*15:03 Frequency Overrepresented in Epidermolysis Bullosa Acquisita

August 2011


120 Reads

Epidermolysis bullosa acquisita (EBA) is a rare autoimmune bullous disease (AIBD). However, higher EBA incidence and predisposing genetic factor(s) involving an HLA haplotype have been suspected in some populations. This retrospective study assessed the overrepresentation of black patients with EBA, its link with HLA-DRB1*15:03, and their clinical and immunological characteristics. Between 2005 and 2009, 7/13 (54%) EBA and 6/183 (3%) other-AIBD patients seen consecutively in our department were black (P=10(-6)); moreover 7/13 (54%) black patients and 6/183 (3%) white patients had EBA (P=10(-6)). In addition, between 1983 and 2005, 12 black patients had EBA. Finally, among the 19 black EBA patients, most of them had very atypical clinical presentations, 9 were natives of sub-Saharan Africa, 1 from Reunion Island, 7 from the West Indies, and 2 were of mixed ancestry. HLA-DRB1*15:03 allelic frequencies were 50% for African patients, significantly higher than the control population (P<10(-3)), and 21% for the West Indians (nonsignificant). High EBA frequencies have already been reported in American blacks significantly associated with the HLA-DR2. In conclusion, black-skinned patients developing EBA seem to have a genetic predisposition, and EBA should be suspected systematically for every AIBD seen in this population.

Significantly Increased Occurrence of HLA-DQB1∗0301 Allele in Patients with Ocular Cicatricial Pemphigoid

February 1997


16 Reads

To determine the immunogenetic characteristics of patients with immune-mediated subepithelial blistering diseases of mucous membranes, we performed HLA-typing for the class II MHC gene DQB1*0301 allele using a direct method. Genomic DNA extracted from Caucasian patients was amplified by polymerase chain reaction using primer pairs specific for the DQB loci followed by Southern blotting with a peroxidase-conjugated sequence-specific oligonucleotide probe. Seventy-six percent (16/21) of patients with ocular mucosal disease (with or without oral mucosal and skin diseases) carried the DQB1*0301 allele; by contrast, only 33% (14/42) of race-, age-, and geography-matched normal individuals carried the DQB1*0301 allele (p < 0.005). The relative risk for ocular disease if DQB1*0301 allele is present is 6.4, similar to the relative risk of 8 for patients with ocular but no oral disease (pure ocular cicatricial pemphigoid, p < 0.025). In patients with oral mucosal disease (with or without ocular mucosal and skin diseases), 68% (15/22) carried the DQB1*0301 allele (p < 0.025). When patients with ocular disease were excluded, however, the increased occurrence of the DQB1*0301 allele in patients with oral disease was not statistically significant (64%, 7/11, p < 0.25). In patients with subepidermal blistering skin disease but no oral or ocular disease, there was no increase in the occurrence of the DQB1*0301 allele (38%, 5/13, p > 0.5). The significantly increased occurrence of the DQB1*0301 allele in patients with ocular mucosal disease may point to a distinct immunogenetic factor that predisposes patients to develop an ocular scarring process.

HLA DQB1*0301 Allele is Involved in the Susceptibility to Erythema Multiforme

November 1991


59 Reads

Erythema multiforme (EM) is an acute, episodic inflammatory disorder of the skin and mucous membranes of various etiology that could he related to immunologic hypersensitivity response. EM has been previously reported to be associated with serologically defined HLA-DRw53 and DQw3 antigens. In this report, we reevaluate the role of HLA class II alleles in EM manifestations. With use of the polymerase chain reaction, followed by sequence-specific oligonucleotide by- hybridization, 35 unrelated Caucasian EM patients and 80 randomly selected healthy subjects were studied, and the DRB3, DRB4, DQA1, and DQB1 alleles were analyzed. The comparison of frequencies of these alleles indicates that (i) susceptibility to EM disease is more associated with the HLA-DQ than tile HLA-DR subregions and (ii) that the DQB1*0301 is the most frequent allele among EM patients. Sixty-six percent of the patients had the DQB1*0301 allele compared to 31% of the controls (RR = 4.1; p<0.001). An even stronger DQB1*0301 association was found in the patient group with herpes-associated EM (76%; RR = 6.5; p < 0.001). Our data demonstrate a clear association between an HLA-DQB1 allele and susceptibility to EM.

HLA-DRB1*04 and DRB1*14 Alleles Are Associated with Susceptibility to Pemphigus Among Japanese

December 1997


28 Reads

It has previously been demonstrated that susceptibility to pemphigus vulgaris is associated with human leukocyte antigen (HLA)-DR4 serologic specificity among Ashkenase Jews, and with DR4 as well as DR6 (DR14) in other ethnic groups. We genotyped HLA-DRB1, DQA1, DQB1, and DPB1 alleles in 16 patients with pemphigus by polymerase chain reaction-restriction fragment length polymorphism, to find evidence of potential HLA class II allele associations with pemphigus in Japanese patients who have a relatively homogeneous ethnic background. All nine patients with pemphigus vulgaris and five of seven patients with pemphigus foliaceus carried one or two alleles of HLA-DRB1*04 (*0403, *0406) and HLA-DRB1*14 (*1401, *1405, *1406) subtypes. Sequence analysis of these DRB1*04 and DRB1*14 alleles revealed the amino acid homology of phenylalanine at position 26 and valine at position 86 with the DRB1*0402 allele that reportedly confers a strong susceptibility to pemphigus vulgaris in Ashkenazi Jews. Thus our findings, together with previous HLA studies on pemphigus vulgaris patients of different ethnic groups, suggest that HLA-DRB1*04 and DRB1*14 alleles are commonly associated with pemphigus vulgaris across racial barriers. These HLA-DRB1 alleles are likely to be also associated with pemphigus foliaceus. Further studies on more diverse ethnic populations will be helpful in determining the significance of the association between certain amino acid residues of the class II molecules and disease susceptibility to pemphigus vulgaris as well as pemphigus foliaceus.

Figure 1: HLA-C*06:02 carrier and noncarrier distribution in the whole sample set. Frequency of HLAC*06:02 allele in the different age-at-onset groups.Download Power Point slide (156 KB)
Table 1 . Characteristics of participants
Figure 2: Association for ERAP1 SNP rs26653 stratified for HLA-Cw06 status and age. Estimates of genotype effects were calculated using logistic regression in the R software package. Individuals with the low-risk genotypes for rs26653 GG and HLA-C*06 NN were set as the baseline. The other genotype combinations were coded according to a series of dichotomous indicator variables. Odds ratios were derived by exponentiation of the relevant coefficient from the logistic regression.Download Power Point slide (173 KB)
Genetic Association with ERAP1 in Psoriasis Is Confined to Disease Onset after Puberty and Not Dependent on HLA-C&ast;06

August 2012


292 Reads

HLA-C remains the strongest susceptibility candidate gene in psoriasis. Evidence for interaction between HLA-C and endoplasmic reticulum aminopeptidase 1 (ERAP1) confined to individuals carrying the HLA-C risk allele was recently reported. Psoriasis displays wide variation, and genetic heterogeneity is likely to contribute to clinical diversity. Age at disease onset is a putative discriminator, and separating psoriasis into early- (<40 years) and late-onset disease has been useful. To sharpen the age-dependent phenotype, we compared genotypes for ERAP1 (rs26653, rs30187, and rs27524) and HLA-C*06:02 in healthy controls and cases stratified for onset of psoriasis at <10, 10-20, 20-40, and >40 years of age. This approach revealed that association with ERAP1 was confined to cases with onset between 10 and 20 years (odds ratio 1.59, 95% confidence interval: 1.28-1.98, P=0.00008) and no association was detected in cases with onset below 10 years, reflecting genetic heterogeneity within the childhood psoriasis population. In contrast to earlier findings, association with ERAP1 was neither dependent on nor interacting with HLA-C*06:02. ERAP1 single-nucleotide polymorphism rs26653, which, to our knowledge, has not previously been reported in psoriasis, is nonsynonymous, has suggestive functional consequences, and herein displays strong association with disease.Journal of Investigative Dermatology advance online publication, 30 August 2012; doi:10.1038/jid.2012.280.

HLA-CW&ast;0602 Is a Susceptibility Factor in Type I Psoriasis, and Evidence Ala-73 Is Increased in Male Type I Psoriatics

September 1997


37 Reads

We investigated the HLA-C locus of 87 unrelated patients with chronic plaque psoriasis by genotyping with sequence-specific amplification primers. The HLA-Cw*0602 allele was significantly increased in male and female type I psoriatics but not significantly increased in either male or female type II psoriatics. The overall frequency of Ala-73 (present in Cw*04, Cw*0602, Cw*07, Cw*12, Cw*1503, and Cw*17) in psoriatics was 88.5% but the incidence of Ala-73 in our Caucasian controls was also high at 84.3%. Ala-73 was present in 97.2% of type I and 85.7% of type II male psoriatics (chi2 = 8.43, p = 0.001; chi2 = 0.01, p = nonsignificant, respectively), in contrast to 81.5% of type I and 80% of type II female psoriatics (nonsignificant). HLA-Cw*0602 appeared more discriminating in determining disease susceptibility in our population than Ala-73, in line with earlier serologic studies implicating HLA-Cw6. Thus, although the frequency of HLA-Cw*0602 decreased from 54.0% in type I to 29.2% in type II psoriatics, the overall frequency of Ala-73, present in 90.4% of type I and 83.3% of type II psoriatics, did not. (i) Thus this study confirms the strong association between psoriasis and HLA-Cw*0602 by using sequence-specific amplification primers. (ii) Results show that Ala-73 on HLA-C molecules is increased in frequency in psoriasis, but results observed show an association more subtle than previously thought, with HLA-Cw*0602 playing the major role. (iii) This report documents the differential association of HLA genes in male and female psoriatic patients. An interaction between gender and immunogenetics may influence susceptibility to psoriasis.

Distinct Clinical Differences Between HLA-Cw*0602 Positive and Negative Psoriasis Patients – An Analysis of 1019 HLA-C- and HLA-B-Typed Patients

April 2006


122 Reads

A major susceptibility gene for psoriasis is located in the major histocompatibility complex class I region on chromosome 6 very close to the HLA-Cw6 gene. We collected a cohort of 1,019 patients with chronic plaque psoriasis. The patients were typed for HLA-C and HLA-B. A total of 654 (64.2%) were HLA-Cw*0602 positive but 365 (35.8%) carried other HLA-C alleles. We confirmed that HLA-Cw*0602 positive patients have younger age of onset (17.5 vs 24.3 years, P<10(-10)), higher incidence of guttate and the eruptive type of psoriasis (P<0.0001), more frequent exacerbations with throat infections (P=0.01), higher incidence of the Koebner's phenomenon (P=0.01), and more extensive disease (P=0.03). A striking new finding was a diverging pattern of disease severity in HLA-Cw*0602 positive and negative patients depending on the age of onset of the disease (P=0.0006). HLA-Cw*0602 positive women also had more frequent remissions during pregnancy (P<0.0001). All types of nail changes were, however, more common in the Cw*0602 negative patients (P=0.003) and they more often had multiple types of nail lesions (P<0.0001). The three ancestral haplotypes of Cw*0602 all conferred an increase in odds ratio but showed no difference in any of the clinical features studied. Our findings indicate that the genetic factor on chromosome 6 has a strong influence on the phenotype of the disease, and underline that differences in clinical features of psoriasis may be to a large extent genetically determined.

The OTF3 Gene Polymorphism Confers Susceptibility to Psoriasis Independent of the Association of HLA-Cw*0602

December 2000


29 Reads

Psoriasis has been strongly associated to HLA-Cw6, but it remains unclear whether Cw6 itself or a closely linked gene is associated with the disease. The aim of this study was to clarify whether the HLA-C itself determines disease susceptibility or whether it acts only as a marker for the susceptibility allele. We examined a sample of 95 type I psoriasis patients and 104 Spanish matched controls to investigate whether HLA-Cw*0602 or other closely related class I loci, such as HLA-B and MICA (which are centromeric to HLA-C), or corneodesmosin gene and octamer transcription factor-3 genes (which are telomeric to HLA-C), might play a part in disease development. DNA samples were genotyped by polymerase chain reaction/sequence-specific primers (HLA-C), polymerase chain reaction/sequence-specific primers (HLA-B), radioactive polymerase chain reaction (MICA-TM polymorphism in the transmembrane region), and polymerase chain reaction/restriction fragment length polymorphism (protein S and octamer transcription factor-3). Our results show a significant increase of Cw*0602 in psoriasis patients (odds ratio = 3.64; pc < 0.0006). A significant association between the beta allele of octamer transcription factor-3 (HindIII) and psoriasis was also detected (odds ratio = 3.76; pc < 0.0003). The allele octamer transcription factor-3B (etiologic fraction = 0.62) was found to be more strongly associated to psoriasis vulgaris than Cw*0602 (etiologic fraction = 0.35) and the increase of octamer transcription factor-3 B allele is independent of the linkage disequilibrium with Cw*0602 as this was also found in Cw*0602 negative patients (odds ratio = 3.63; pc < 0.015, etiologic fraction = 0.55). We did not detect an association between the corneodesmosin gene and psoriasis. This fact suggests that the psoriasis susceptibility gene is located within a critical region of 147 kb, telomeric to HLA-C and centromeric to the corneodesmosin gene, and the association of Cw6 to psoriasis may be secondary to linkage disequilibrium.

Increased Phospholipase C-Catalyzed Hydrolysis of Phosphatidylinositol-4,5-Bisphosphate and 1,2-sn-Diacylglycerol Content in Psoriatic Involved Compared to Uninvolved and Normal Epidermis

November 1990


20 Reads

Evidence suggests that the phospholipase C/protein kinase C signal transduction system participates in the regulation of epidermal cell growth and differentiation. Psoriatic epidermis is characterized by hyperproliferation, defective differentiation, and inflammation. In this report, we have determined the activity of phospholipase C-catalyzed hydrolysis of phosphatidylinositol-4,5-bisphosphate (PIP2) and 1,2-diacylglycerol content in normal and psoriatic involved and uninvolved epidermis. 1,2-diacylglycerol is formed from phospholipase C-catalyzed hydrolysis of PIP2 and is the physiologic activator of protein kinase C. PIP2 hydrolysis was assayed in soluble and particulate fractions prepared from keratome biopsies of normal and psoriatic skin. Total lipids were extracted from normal and psoriatic epidermis and 1,2-diradylglycerol (a mixture of 1,2-diacylglycerol and 1-ether, 2-acyl-glycerol) quantitated by enzyme assay. Because 1,2-diacylglycerol is a more potent activator of protein kinase C, the relative proportions of 1,2-diacyl and 1-ether, 2-acylglycerol in uninvolved and involved psoriatic epidermis were determined. This was accomplished by separation of acetate derivatives of 1,2-diacylglycerol and 1-ether, 2-acyl-glycerol by thin layer chromatography. Soluble and membrane-associated phospholipase C-catalyzed PIP2 hydrolysis were increased 3.7 times (p less than 0.001) and 3 times (p less than 0.004), respectively, in psoriatic involved compared to uninvolved and normal epidermis. 1,2-diradylglycerol content was also significantly elevated (3 times, p less than 0.01) in psoriatic involved versus uninvolved and normal epidermis. Analysis of the acetate derivatives of 1,2-diradylglycerol in psoriatic uninvolved and involved epidermis revealed that 1,2-diacylglycerol was the major species (86% and 95%, respectively). There were no significant differences in either phospholipase C-catalyzed PIP2 hydrolysis or 1,2-diacylglycerol content between uninvolved and normal epidermis. 1,2-diacylglycerol purified from normal and involved psoriatic epidermis was capable of activating protein kinase C from normal epidermis in vitro. In epidermal slices, activation of protein kinase C by addition of 12-0-tetradecanoylphorbol-13-acetate and 1,2-diacylglycerol (1,2-dioctanoylglycerol) resulted in subsequently decreased protein kinase C activity, a process termed down-regulation. These data are consistent with the possibility that the elevation in lesional 1,2-diacylglycerol content may account, in part, for the previously reported reduction of protein kinase C activity in psoriasis (Horn, Marks, Fisher, et al: J Invest Dermatol 88:220-222, 1987).

Agonist-induced Hydrolysis of Phosphoinositides and Formation of 1,2-Diacylglycerol in Adult Human Keratinocytes

September 1989


23 Reads

The ability of a variety of agonists to induce formation of inositol phosphates and 1,2-diacylglycerol in cultured adult human keratinocytes has been investigated. Histamine, bradykinin, and thrombin significantly stimulated formation of inositol mono-, bis-, and trisphosphate and 1,2-diacylglycerol within 5 min after addition. Aluminum fluoride also caused a dose-dependent accumulation of inositol phosphates suggesting the participation of a GTP binding protein in the regulation of phospholipase C-catalyzed phosphoinositide hydrolysis. These data demonstrate that human keratinocytes possess the capacity for phospholipase C-mediated signal transduction and suggest that this pathway may participate in the regulation of keratinocyte function.

Top-cited authors