Jean Philippe Lacour

Defective Expression of Plectin/HD1 in Epidermolysis Bullosa Simplex with Muscular Dystrophy

Epidermolysis bullosa simplex with muscular dystrophy (MD-EBS) is a disease characterized by generalized blistering of the skin associated with muscular involvement. We report that the skin of three MD-EBS patients is not reactive with antibodies 6C6, 10F6, or 5B3 raised against the intermediate filament-associated protein plectin. Immunofluorescence and Western analysis of explanted MD-EBS keratinocytes confirmed a deficient expression of plectin, which, in involved skin, correlated with an impaired interaction of the keratin cytoskeleton with the hemidesmosomes. Consistent with lack of reactivity of MD-EBS skin to plectin antibodies, plectin was not detected in skeletal muscles of these patients. Impaired expression of plectin in muscle correlated with an altered labeling pattern of the muscle intermediate filament protein desmin. A deficient immunoreactivity was also observed with the monoclonal antibody HD121 raised against the hemidesmosomal protein HD1. Furthermore, immunofluorescence analysis showed that HD1 is expressed in Z-lines in normal skeletal muscle; whereas this expression is deficient in patient muscle. Colocalization of HD1 and plectin in normal skin and muscle, together with their impaired expression in MD-EBS tissues, strongly suggests that plectin and HD1 are closely related proteins. Our results therefore provide strong evidence that, in MD-EBS patients, the defective expression of plectin results in an aberrant anchorage of cytoskeletal structures in keratinocytes and muscular fibers leading to cell fragility.

Multicenter prospective study of the humoral autoimmune response in bullous pemphigoid

Bullous pemphigoid (BP) is an autoimmune bullous disease, associated with autoantibodies directed against the hemidesmosomal components BP180 and BP230. In this study for the first time different laboratories have analyzed the autoantibody profile in the same group of 49 prospectively recruited BP patients. The results show that: 1) disease severity and activity correlated with levels of IgG against the BP180-NC16A domain, but also against a COOH-terminal epitope of BP180, 2) distinct epitopes of the BP180 ectodomain other than BP180-NC16A were recognized by 96% of the BP sera; and 3) the combined use of BP180 and BP230 ELISA led to the detection of IgG autoantibodies in all the BP sera. These results demonstrate the usefulness of the combined ELISAs based on various BP180 and BP230 fragments in establishing the diagnosis of BP and support the concept that BP180 is the major autoantigen of BP.

Demonstration of Epitope-Spreading Phenomena in Bullous Pemphigoid: Results of a Prospective Multicenter Study

Bullous pemphigoid (BP), the most common autoimmune subepidermal bullous disease, is associated with an autoantibody response to BP180 and BP230, two components of junctional adhesion complexes in human skin promoting dermo-epidermal cohesion. Retrospective analyses demonstrated that these autoantigens harbor several epitopes targeted by autoaggressive B and T cells. The aim of this prospective multicenter study was to assess the evolution of IgG autoantibodies in 35 BP patients over a 12-month observation period. Epitope-spreading (ES) events were detected in 17 of 35 BP patients (49%). They preferentially occurred in an early stage of the disease and were significantly related to disease severity at diagnosis. Moreover, in three patients, spreading of IgG reactivity to intracellular epitopes of BP180 and BP230 was preceded by recognition of the BP180 ectodomain. Finally, IgG reactivity with extracellular epitopes of BP180 and intracellular epitopes of BP230 correlated with the severity of BP in disease course. These findings support the idea that IgG recognition of the BP180 ectodomain is an early and crucial event in BP disease, followed by variable intra- and intermolecular ES events, which likely shape the individual course of BP.

Immunofluorescence Analysis of Villous Trophoblasts: A Tool for Prenatal Diagnosis of Inherited Epidermolysis Bullosa with Pyloric Atresia

Genetic mutations invalidating the genes for integrin alpha6beta4 and, in some cases, plectin are associated with junctional and simplex epidermolysis bullosa with pyloric atresia (PA-JEB and PA-EBS), respectively. These recessive inherited conditions are characterized by pregnancies with fetal bullae, pyloric atresia, polyhydramnios, and neonatal mucocutaneous blistering, which often results in early postnatal demise. To date, first-trimester DNA-based prenatal diagnosis is not applicable to affected kindred carrying as yet unidentified genetic mutations. Here, we show that first-trimester chorionic villi strongly express both integrin alpha6beta4 and plectin, which persist throughout the pregnancy. Based on this observation, we implemented 25 prenatal diagnoses in kindred at risk for PA-EB by immunomapping, which identified three PA-JEB-affected fetuses and 22 healthy ones. In 19 cases, including the three PA-JEB pregnancies that were prematurely terminated, the results were confirmed by chorionic villous DNA-based tests, which also led to the identification of seven previously unreported mutations in the alpha6beta4 integrin genes. Our prediction was further sustained by the birth of 22 healthy babies. These results validate chorionic villi immunofluorescence examination as a tool for prenatal diagnosis of PA-JEB and PA-EBS and indicate that this procedure could be devised for EB with muscular dystrophy, which is also associated with genetic mutations in plectin.

No correlation between the molecular subtype of COL1A1-PDGFB fusion gene and the clinico-histopathological features of dermatofibrosarcoma protuberans.

TO THE EDITOR Dermatofibrosarcoma protuberans (DFSP) is a dermal tumor of intermediate malignancy. Propensity for metastases is low but local recurrence risk is high unless a wide surgical excision is performed. In addition to the classical presentation of DFSP, several clinical and histological variants have been described, such as DFSP-containing fibrosarcomatous areas or malignant fibrous histiocytoma areas, Bednar tumor, giant cell fibroblastoma, and superficial adult fibrosarcoma (Simon et al., 1997; Sheng et al., 2001; Maire et al., 2002a; Sirvent et al., 2003; Bianchini et al., 2007; Szollosi et al., 2007). DFSP and related tumors are molecularly characterized by the presence of an abnormal chimeric gene that fuses COL1A1 (17q21.3) with platelet-derived growth factor-B chain (PDGFB) (22q13.1) (Simon et al., 1997). The detection of COL1A1– PDGFB is mandatory in case of unusual clinical or histological presentation, as well as in pediatric cases (Sirvent et al., 2003; Bianchini et al., 2007; Maire et al., 2007). A striking feature of COL1A1–PDGFB fusion consists of the high variability of the break point location between the exons 6 and 49 of COL1A1 (Sirvent et al., 2003; Bianchini et al., 2007). In contrast, the break point in PDGFB is consistently located in the first intron. We have investigated whether the COL1A1 break point location is associated with clinical or histological features. In such a case, the precise detection of the COL1A1 break point detection by using multiplex reverse transcriptase–PCR and sequencing could be useful for diagnosis or evaluation of prognosis. For this purpose we compiled a series of 172 DFSP cases with both molecular and clinico-histological data (Supplementary Table S1), including (i) 35 cases of DFSP or related tumors addressed for molecular diagnosis referral to the laboratory of Solid Tumors Genetics (Nice, France) from nine different French Hospitals and previously unreported; (ii) 22 DFSP cases previously published by our group (Pedeutour et al., 1994, 1995, 1996; Simon et al., 1997; Greco et al., 1998; Navarro et al., 1998; Maire et al., 2002a, b, c, 2007; Terrier-Lacombe et al., 2003; Jouary et al., 2007); (iii) 115 cases published between 1997 and 2008 (O’Brien et al., 1998; Wang et al., 1999, 2000; Vanni et al., 2000; Sheng et al., 2001; Gokden et al., 2003; Sandberg et al., 2003; Sirvent et al., 2003; Saeki et al., 2003a, b, 2005, 2006; Martin et al., 2005; Craver et al., 2006; Kashima et al., 2006; Llombart et al., 2006, 2008; Nakanishi et al., 2007; Szollosi et al., 2007; Takahira et al., 2007; Patel et al., 2008). The description of the clinical and histological features of the 172 cases is compiled in Supplementary Table S1. Continuous data (age, tumor size) were described with mean, SD, and range (minimum–maximum); nominal scale data (gender, anatomic location, histological variant) were described by frequencies and relative frequencies (expressed as percentage). A Fisher exact test showed no significant differences in terms of gender, clinical presentation, and histological variant between our series and tumors studied by other groups (Table 1). Similarly, an unpaired Student’s t-test showed no significant differences in the two series in terms of tumor size. In contrast, a slight difference between the two series was observed for the anatomic location and the mean age. The first could be explained by reports of exceptional anatomical locations in the literature, and the later by a more frequent recruitment of pediatric patients in our laboratory. The very high variability of the COL1A1 break point position was confirmed, as 38 different COL1A1 exons have been identified. The most frequently rearranged COL1A1 exons were exon 25 (n1⁄4 19), 32 (n1⁄418), and 47 (n1⁄4 15) (Figure 1). A standard analysis of variance of the COL1A1 break point position with respect to the clinico-histological parameters was performed by grouping the break points into four distinct groups of 11 exons each (group A: exons 6–16; group B: exons 17–27; group C: exons 28–38; and group D: exons 39–49). The following parameters were tested for the COL1A1 break point position: age, histological variant, anatomic location, and gender. None of the parameters tested showed a significant association with the either group. Only the anatomic location showed a slight trend, P1⁄40.01843 (Supplementary Table S2). Also, no significant correlation between the most represented COL1A1 break points (n45) (exons 7, 25, 29, 32, 33–34, 40, 42, 46, and 47) and clinical or histological features was observed. This result is consistent with the role of the COL1A1 side of the fusion gene as a simple cis element for PDGFB overexpression (Greco et al., 1998; Simon et al., 2001). The spectacular clinical effect of tyrosine kinase receptor inhibitor (imatinib mesylate) also supports the major role of the PDGFR activation through the overexpression of PDGFB Abbreviations: COL1A1, collagen type I-a 1; DFSP, dermatofibrosarcoma protuberans; PDGFB, plateletderived growth factor-B chain

In vivo reflectance confocal microscopy of the skin: A noninvasive means of assessing body cystine accumulation in infantile cystinosis

BACKGROUND Patients with infantile nephropathic cystinosis have progressive accumulation of cystine in tissues leading to delayed extrarenal complications. No simple tool is available to evaluate the level of body cystine accumulation. OBJECTIVE We sought to determine the value of in vivo reflectance confocal microscopy of the skin in patients with infantile nephrogenic cystinosis. METHODS Nine patients and control subjects were recruited for this study. Images were acquired by means of a near-infrared reflectance confocal laser scanning microscope. RESULTS Scattered bright particles within the papillary dermis were observed in all patients but not in control subjects. The density of particles ranged from numerous (+++) to very few (+/-) and their distribution was heterogeneous. Electron microscopy confirmed that these particles corresponded to cystine crystal deposits within dermal fibroblasts. The density of cystine crystals within the dermis was greater in older patients, in patients with a high leukocyte cystine concentration, and with delayed cysteamine therapy. There was no correlation between the density of cystine deposits and renal disease or hypopigmentation but high levels of deposition occurred in association with extrarenal manifestations. LIMITATIONS This is a preliminary study on a small sample of patients. Repeated examination and longer follow-up is necessary. CONCLUSION In vivo reflectance confocal microscopy of the skin appears to be a noninvasive means of assessing body cystine accumulation in infantile cystinosis and could be used as a complementary marker of treatment response in addition to leukocyte cystine measurement.

Epidermolysis Bullosa Acquisita With Negative Direct Immunofluorescence

Epidermolysis bullosa (EB) appeared in a patient at the age of 54 years. Other bullous disorders could be excluded by electron microscopy, and there was no family history of EB. The patient would therefore best be classified as having EB acquisita. Repeated direct immunofluorescence studies were, however, negative for all tested serum samples, suggesting that there might be a subgroup lacking immunoglobulin deposits in the skin. Collagen IV, laminin, and fibronectin were expressed normally at the dermoepidermal junction.

Neutrophilic dermatosis associated with chronic neutrophilic leukemia

Chronic neutrophilic leukemia is an uncommon myeloproliferative disorder. We report a new case that fulfills the clinical and biologic criteria for such a diagnosis. The hematologic disease was revealed by a neutrophilic dermatosis that finally disappeared spontaneously after a duration of 1 year. Despite the lack of parallelism in the course of dermatologic and hematologic manifestations, we believe they were strongly linked. Occurrence of neutrophilic dermatoses in the course of other myeloproliferative disorders is well known. However, in our case, clinical and histologic features could not be used to distinguish between atypical Sweets syndrome and specific cutaneous lesions because of the mature appearance of both skin and blood neutrophils.

Macular eruption revealing hypomelanotic cutaneous melanoma metastases: Diagnostic role of dermoscopy

the Department of Dermatology, INSERM U 1065, Team 1, d the Clinical Research Centre, University Hospital of Nice; HU Nice, Laboratory of Clinical and Experimental Pathology, steur Hospital, Nice; and the Department of Head and Neck rgery, Cancer Center Antoine-Lacassagne, University Nice phia-Antipolis, Nice. ing sources: None. licts of interest: None declared. Reprint requests: Philippe Bahadoran, MD, PhD, Department of Dermatology, University Hospital of Nice, Archet 2 Hospital, 151 Rte de St-Antoine de Ginestiere, 06200 Nice, France. E-mail: bahadoran.p@chu-nice.fr. J Am Acad Dermatol 2014;70:e7-9. 0190-9622/

Cutaneous Leishmaniasis Due to Leishmania infantum: Case Reports and Literature Review

36.00 a 2013 by the American Academy of Dermatology, Inc. http://dx.doi.org/10.1016/j.jaad.2013.09.004