The FAS/cd95 promoter single-nucleotide polymorphism -670 A/G and lupus erythematosus.

An increased level of circulating nuclear antigens caused by apoptosis is thought to be responsible for the production of autoantibodies in lupus erythematosus (LE). The presentation of these antigens to immunologically competent cells may trigger systemic autoimmunity. The influence of a functional single-nucleotide polymorphism at position -670 in the promoter of the apoptosis gene FAS on susceptibility to autoimmune diseases including systemic LE has been a controversial subject. Although it has not yet been possible to assign any particular allele or genotype to the control of FAS expression, this polymorphism has been described to be associated with several autoimmune diseases including LE. When we compared the FAS -670 A/G genotypes of 107 German patients with LE and those of 96 healthy controls, we found a trend for association between LE and the homozygous A genotype in the patient group. This finding suggests that apoptosis may contribute to development of autoimmune reactions and that FAS function might be relevant for LE.

Lupus erythematosus tumidus is a separate subtype of cutaneous lupus erythematosus.

Background Lupus erythematosus tumidus (LET) is a rare disease which was first described in 1909 but has not always been considered as a separate entity of cutaneous lupus erythematosus (CLE) in the international literature. Objectives To compare characteristic features of different subtypes of CLE and to analyse whether LET can be distinguished as a separate entity in the classification system of the disease. Methods The study involved 44 patients with CLE, including 24 patients with LET, 12 with discoid lupus erythematosus (DLE) and eight with subacute CLE (SCLE), from two centres in Germany. A core set questionnaire and an SPSS database were designed to enable a consistent statistical analysis. Results Location of skin lesions did not differ significantly between the CLE subtypes; however, the activity score was significantly lower in LET than in DLE (P < 0.01), and the damage score was significantly lower in LET than in SCLE (P < 0.01) and DLE (P < 0.01). Photosensitivity and antinuclear antibodies were confirmed to be different in LET compared with SCLE and DLE but without statistical significance. Moreover, histological analysis of skin biopsy specimens showed that abundant mucin deposition is significantly more present in LET compared with SCLE (P < 0.01) and DLE (P < 0.01) while prominent interface dermatitis and alteration of hair follicles were absent in LET. Conclusions Several significant differences were found between LET and other subtypes of CLE with regard to clinical, histological and laboratory parameters. These data strongly indicate that LET should be defined as a separate entity in the classification of CLE.

Confocal laser-scanning capillaroscopy: a novel approach to the analysis of skin capillaries in vivo.

BACKGROUND: New techniques for diagnostics and therapy in dermatology are becoming increasingly non-invasive, among which confocal laser-scanning microscopy (CLSM) is the most prevalent. It allows visualization of cellular structures of the skin up to a depth of 300 microm in vivo. Until now, most studies have been conducted on pathologically altered skin, mostly oncologic lesions. We now present a detailed analysis of capillaries located in the upper dermal papillae. METHODS: Multiple measurements were performed on the dorsal and ventral surface of the right forearm of 30 healthy volunteers (22-88 years) under standard conditions (room temperature, body position, time of day). Images were obtained with the Vivascope 1500 (Lucid) under standard settings and analyzed using the freeware ImageJ with a customwritten macro plugin. The following parameters of the capillaries in vivo were measured: area, perimeter, circularity and maximum diameter. RESULTS: Statistical analysis showed that all four parameters were constant within a narrow range, regardless of the body site, sex and age. In this physiological study, we can clearly demonstrate that by confocal laser-scanning capillaroscopy (CLSC), it is possible to visualize and measure skin capillaries at the extremities in a reproducible manner. CONCLUSION: This new approach offers a considerable advantage compared with nailfold capillaroscopy, which can only be performed at the proximal nail segment, and over histological analysis, which can be hampered by fixation artifacts resulting in altered size and shape of the vessels to be analyzed. CLSC could allow for precise analysis of in vivo skin vasculature in systemic and proliferative diseases of the skin.

In a study for acne vulgaris, sequence-based HLA typing showed a novel DPB1 allele, DPB1*2402.

In this paper, we characterize the novel human leucocyte antigen (HLA)-DPB1*2402 allele that we found in a patient suffering from acne vulgaris. In comparison to the closest related allele DPB1*0401, HLA-DPB1*2402 has a single nucleotide exchange at position 115 (202), T replaces G. In consequence, codon 39 (68) TAC encodes for tyrosine in the novel allele instead of aspartic acid 39 (68) GAC in DPB1*0401.

[Autoantibody detection by indirect immunofluorescence on HEp-2 cells]. / Autoantikörpernachweis mittels indirekter Immunfluoreszenz an HEp-2-Zellen.

Systemic autoimmune diseases are characterized by the presence of antinuclear autoantibodies (ANA). Diluted patient sera are typically used to screen for the presence of ANA by immunfluorescence microscopy with fixed HEp-2 cells. Despite high-quality test kits, reports of different laboratories frequently present controversial results. This article recommends unified processing and interpretation of HEp-2 based screening for autoantibodies. Suggestions are made for the selection of high-quality test kits, optimized processing and diagnostic procedures. In addition to a relevant clinical diagnosis and an experienced laboratory specialist, the following procedure is highly recommended to achieve good laboratory practice: Initial HEp-2 based screening by indirect immunofluorescence, starting with a 1:80 serum dilution, and evaluation in a bright fluorescence microscope, pathological values from a titer of 1:160 upwards, internal quality checks and unified interpretation. We aim to improve diagnosis and care of patients with autoimmune diseases as a central focus of the European Autoimmunity Standardization Initiative (EASI).

Autoantibodies against desmocollins in European patients with pemphigus.

BACKGROUND: Autoimmune bullous disorders of the pemphigus group are characterized by autoantibodies targeting desmoglein (Dsg)1, Dsg3 and Dsg4 and occasionally, desmocollin (Dsc)1, Dsc2 and Dsc3. Both Dsg and Dsc are components of desmosomal adhesion complexes. AIM: To investigate the presence of IgG and IgA autoantibodies against Dsc1, Dsc2 and Dsc3 in a cohort of patients with bullous disorders. METHODS: IgG and IgA autoantibodies against Dsc1, Dsc2 and Dsc3 were investigated by ELISA and immunoblotting analysis in a cohort of European patients with pemphigus vulgaris (PV; n = 74), IgA pemphigus (n = 3), paraneoplastic pemphigus (PNP; n = 3) and two cases of atypical pemphigus (n = 2). RESULTS: Of the two cases with atypical pemphigus, one showed IgA reactivity against Dsc1 and Dsc3 and weak reactivity against Dsc2, and the other showed both IgG and IgA reactivity against Dsc1. One patient with IgA pemphigus had IgA autoantibodies against Dsc1, Dsc2 and Dsg1, and one patient with PNP had IgG reactivity against with Dsc3. In contrast, all the PV sera showed IgG reactivity against Dsg3 but not against Dsc1-3. Thus, IgG and IgA reactivity against Dsc was restricted to cases of PNP, IgA pemphigus and atypical pemphigus. CONCLUSIONS: These findings support the concept that desmocollins are not important autoantigens in PV.

Relevance of the low-affinity type of the Fcgamma-receptor IIIa-polymorphism in bullous pemphigoid.

Bullous pemphigoid (BP) is mediated by autoantibodies directed against molecules of the basement membrane zone. The biological function of antibodies involves binding to Fc-receptors expressed on human leucocytes. Recent studies suggested that a functional single-nucleotide-polymorphism of the Fcgamma-receptor IIIa (FcgammaRIIIa = CD16) at nucleotide 559 might predispose to the development of antibody-associated autoimmune disorders. This allelic difference affects the level of receptor affinity by predicting either a phenylalanine (F 158, low-affinity) or valine (V 158, high-affinity). We investigated if inherited frequencies of the high- and low-affinity FcgammaRIIIa polymorphism differed between patients with BP and healthy subjects. Genomic DNA from peripheral white blood cells was analyzed regarding FcgammaRIIIa polymorphism at nucleotide 559 by an established polymerase chain reaction. Sixty-seven Caucasian patients with BP and 88 healthy controls were included into the study. There was no significant difference in the distribution of the homozygous high-affinity FcgammaRIIIa-allotype (V/V) between BP-patients (14.9%) and healthy control subjects (20.5%). In contrast, 58.2% of the BP-patients were homozygous for the low-affinity FcgammaRIIIa-allotype (F/F), compared to 28.4% of the healthy controls (P = 0.001, OR 3.51). The frequencies of the polymorphism in the control group were in range of formerly published frequencies for healthy Caucasian subjects. Thus, the FcgammaRIIIa (158 F/V) polymorphism may modulate the susceptibility to acquire BP.

Shear in flexure of fiber composites with different end supports.

The integrity of fiber-reinforced composite (FRC) prostheses is dependent, in part, on flexural rigidity. The object of this study was to determine if the flexure behavior of uniform FRC beams with restrained or simply supported ends and various length/depth (L/d) aspect ratios could be more accurately modeled by correcting for shear. Experimental results were compared with three analytical models. All models were accurate at high L/d ratios, but the shear-corrected model was accurate to the lowest, more clinically relevant, L/d values. In this range, more than 40% of the beam deflection was due to shear.

Cicatricial pemphigoid: IgA and IgG autoantibodies target epitopes on both intra- and extracellular domains of bullous pemphigoid antigen 180.

BACKGROUND: Cicatricial pemphigoid (CP) is an autoimmune subepidermal blistering disease where autoantibodies target various components of the dermal-epidermal junction, including the bullous pemphigoid antigen 180 (BP180). OBJECTIVE: We determined the exact specificity of circulating IgG and IgA autoantibodies to BP180 in a large number of CP patients. METHODS: Twenty-six consecutive CP sera were analysed by Western blotting using a panel of cell-derived and recombinant proteins covering the entire BP180 molecule. RESULTS: Circulating autoantibodies were detected in all CP sera. Seven sera reacting with laminin-5 were excluded from further analyses; the remaining 19 sera recognized BP180, including six sera (32%) that showed only IgA reactivity to this protein. With the combined use of the soluble BP180 ectodomain (LAD-1) and recombinant BP180 NC16A, 16 of these 19 CP sera (84%) targeted BP180. IgG reactivity was preferentially found against NC16A, whereas IgA antibodies predominantly recognized LAD-1. Thirty-two per cent of the BP180-reative sera revealed reactivity with the intracellular domain of this protein. CONCLUSIONS: Our findings demonstrate that autoantibodies in CP target epitopes on both extra- and intracellular domains of BP180 and highlight the importance of testing for both IgG and IgA reactivity in these patients' sera.

Morphological and biochemical properties of alveolar fibroblasts in interstitial lung diseases.

The phenotype of alveolar-associated fibroblasts (Afb) in sarcoidosis (SA) and idiopathic pulmonary fibrosis (IPF) is unclear. In the present study, we characterized the cytoskeletal proteins and the contraction properties in alveolar-associated fibroblasts recovered by bronchoalveolar lavage (BAL) in the two diseases. Afb were studied from BAL cells in eight IPF and seven SA patients. Cytoskeletal proteins were identified by ELISA and immunofluorescent methods. Biochemical measurements were done by dry chemistry. Contraction was performed by a gel contraction assay. Afb alpha-SM actin measured by ELISA was higher in IPF than in SA (p = 0.042). Vimentin, desmin, myosin, and fibroblast markers were expressed equally. Only in IPF did the Afb reveal the myofibroblast phenotype showing alpha-SM actin immunofluorescence labeling and, by electron microscopy, filaments with associated dense bodies with rough endoplasmic reticulum. Gel contraction showed that cells in IPF contracted significantly more than in SA (p = 0.046 IPF versus SA). The addition of ET-1 increased contraction in all groups. Dry chemistry analysis showed higher levels (p = 0.0065) of creatine phosphokinase (CPK), lower levels of glucose (p = 0.0082), and similar levels of Ca(2+) and lactate in the IPF and SA Afb. Dinitrofluorobenzene (DNFB), a potent inhibitor of CPK, completely abolished spontaneous cell contraction. Afb differentiates into myofibroblasts with different biochemical and energetic properties in IPF. Moreover, Afb from IPF patients showed increased contractile properties. This may explain the difference in the behavior patterns and outcomes of the two diseases.

[Glucocorticoid-induced osteoporosis. A underestimated sequela of long-term treatment of autoimmune diseases]. / Glukokortikosteroidinduzierte Osteoporose. Eine zu wenig beachtete Folge der langfristigen Behandlung von Autoimmunerkrankungen.

The treatment of autoimmune diseases with systemic glucocorticosteroids remains a therapeutic challenge and requires close collaboration with internists, radiologists and in some cases orthopedic surgeons. Generally, patients initially receive high-dose glucocorticosteroid therapy and are then treated for a longer period of time with moderate to low doses above the level causing symptoms of Cushing's disease. A major cause of complications is glucocorticosteroid-induced osteoporosis with hip fractures, crush fractures of the spine and other low trauma fractures as well as deformities of the skeleton leading to neurological and other systemic problems. Loss of bone mass as a result of high dose or long term systemic glucocorticosteroid treatment is well studied and can be documented in a standardized and reproducible fashion using modern radiological techniques. In recent years several controlled studies of bone loss and therapy of osteoporosis have been published, mostly including patients with rheumatoid arthritis or systemic lupus erythematosus. This review discusses recent publications and provides a brief overview on therapeutic options.

[Pemphigus vegetans. A historical perspective]. / Pemphigus vegetans. Eine historische Betrachtung.

Pemphigus vegetans is a rare variant of pemphigus which is mainly localized in the intertriginous areas (Neumann type, Hallopeau type) or primarily involves the mucous membranes (pyostomatitis vegetans). A 18-year-old patient with erosive stomatitis developed a vegetating plaque with papillomatous and verruciform features in her left axilla. Histopathological examination of the axilla revealed papillomatosis and acanthosis as well as suprabasal clefting with acantholytic cells. By direct immunofluorescent examination, deposits of immunoglobulin IgG and complement (C3) were found in the intercellular space of the epidermis. Serological examination by indirect immunofluorescent techniques was indicative of pemphigus autoantibodies at a titer of 1:40. This case report demonstrates that the classical differentiation of pemphigus vegetans in two types, Neumann type and Hallopeau type, is mainly of historical importance, because both entities may represent variants of the same disease. In addition, other disorders such as IgA pemphigus may also present with vegetating plaques in intertriginous sites.

Mapping of epitopes on the BP180 ectodomain targeted by IgA and IgG autoantibodies in patients with the lamina lucida-type of linear IgA disease.

Linear IgA disease (LAD) is an autoimmune subepidermal blistering skin disease characterized by the linear deposition of IgA at the dermoepidermal junction. Serum from patients with LAD most commonly contains autoantibodies that are directed against the hemidesmosomal transmembrane glycoprotein BP180 (type XVII collagen). Various antigenic sites on the extracellular domain of this anchoring filament protein have been shown to be targeted by autoantibodies in different autoimmune bullous skin diseases, including bullous pemphigoid and cicatricial pemphigoid (CP). However, little is known about epitopes on BP180 recognized by autoantibodies in LAD. In this study, we used three recombinant GST fusion proteins, together roughly covering the entire BP180 ectodomain, to characterize the autoimmune response in serum from patients with LAD. Interestingly, we found both IgA and IgG reactivity to all three portions of the BP180 ectodomain. The strongest reactivity was observed with the C-terminal portion of BP180. This is also the major region recognized by autoantibodies in patients with CP. This finding correlates with the observation that there may be significant overlap of the clinical and immunopathological findings in LAD and CP.

Quantitation of human herpes virus 8 DNA in paraffin-embedded biopsies of HIV-associated and classical Kaposi's sarcoma by PCR.

BACKGROUND: Kaposi's sarcoma occurs in patients seropositive and seronegative for the human immunodeficiency virus (HIV) and has been associated with human herpes virus 8 (HHV8). The purpose of this study was to determine and to compare the amount of HHV8 DNA in formalin-fixed tissue sections of Kaposi's sarcoma. METHODS: From 27 biopsies of Kaposi's sarcoma patients, tissue sections were taken and deparaffinized. Four patients were HIV seronegative and 13 were HIV seropositive. After extraction of DNA copy numbers of HHV8 and beta-globin were determined in every sample by quantitative PCR ELISA using an internal quantitation standard. Results were expressed as HHV8 per beta-globin. RESULTS: No significant differences were found between biopsies from HIV-positive and HIV-negative patients (14.8+/-19.6 HHV8 per 1000 beta-globin in HIV-positive versus 18.0+/-23.5 in HIV-negative patients). CONCLUSIONS: These data suggest that HHV8 viral load in Kaposi's sarcoma is relatively low and does not differ in HIV-positive and HIV-negative samples. The importance of viral load determination for prognosis or treatment monitoring remains to be elucidated.

[Bullous scabies and scabies-triggered bullous pemphigoid]. / Bullöse Skabies und durch Skabiesbefall getriggertes bullöses Pemphigoid.

Two patients were admitted to our hospital with tense blisters on an erythematous base, typical for bullous pemphigoid. In both patients an infestation with Sarcoptes scabiei was diagnosed by dermatoscopy as well as histological examination. In one patient the clinical diagnosis of bullous pemphigoid could be confirmed by immunofluorescence microscopy, histopathology and a clinical relapse of bullous pemphigoid without scabies infestation. In the other patient no evidence for an autoantibody-mediated autoimmune blistering disease was found. We postulate that bullous scabies could develop after long persistency of the parasites leading to a specific immune response with activation of T helper type 2 (Th2) cells causing high levels of the cytokine interleukin 5 and then consecutively eosinophilia. Secretion of proteolytic enzymes near the basal membrane zone might explain the development of intraepidermal, often suprabasal blisters. In contrast, in the first patient the scabies infestation might have triggered a flare up of the underlying autoimmune disease. Comparison of our two patients demonstrates two entities: bullous pemphigoid triggered by scabies as a Koebner phenomenon and a bullous subtype of scabies mimicking bullous pemphigoid. Therefore both, scabies infestation triggering bullous pemphigoid and bullous pemphigoid-like scabies should be included in the differential diagnosis of vesicles, tense blisters and erythema, especially at an early clinical stage.