A Slowly Growing Orange Patch on the Cheek: Diagnosis of Lupus Vulgaris 20 Years After Onset of First Skin Changes

<p><b>Article full text</b></p> <p><br></p> <p>The full text of this article can be found here<b>. </b><a href="https://link.springer.com/article/10.1007/s13555-016-0158-x">https://link.springer.com/article/10.1007/s13555-016-0158-x</a></p><p></p> <p><br></p> <p><b>Provide enhanced content for this article</b></p> <p><br></p> <p>If you are an author of this publication and would like to provide additional enhanced content for your article then please contact <a href="http://www.medengine.com/Redeem/”mailto:[email protected]”"><b>[email protected]</b></a>.</p> <p><br></p> <p>The journal offers a range of additional features designed to increase visibility and readership. All features will be thoroughly peer reviewed to ensure the content is of the highest scientific standard and all features are marked as ‘peer reviewed’ to ensure readers are aware that the content has been reviewed to the same level as the articles they are being presented alongside. Moreover, all sponsorship and disclosure information is included to provide complete transparency and adherence to good publication practices. This ensures that however the content is reached the reader has a full understanding of its origin. No fees are charged for hosting additional open access content.</p> <p><br></p> <p>Other enhanced features include, but are not limited to:</p> <p><br></p> <p>• Slide decks</p> <p>• Videos and animations</p> <p>• Audio abstracts</p> <p>• Audio slides</p

Determination of p53 reporter activities associated with cell cycle arrest.

<p>The indicated cells stably transduced with the pGreenFire reporter construct encoding luciferase and GFP under the control of a p53 reporter were cultured in the absence or presence of the mdm-2 inhibitor Nutlin 3a. After 24 hours p53 expression (C), luciferase acitvity (A) and GFP fluorescence (B) were analyzed. Cell cycle analysis was performed on day 2 of treatment and the percentage of cells in S-phase are depicted (D).</p

Functional inactivity in 4 out of 8 melanoma cell lines expressing wild type p53 measured by the p53 reporter construct pGreenFire®.

<p>Human epidermal melanocytes and the indicated melanoma cell lines were stably transduced with a lentiviral pGreenFire reporter construct encoding luciferase and GFP under the control of a p53 responsive element (4× CGACATGCCCGGGCATGT). The cells were then infected with lentiviral supernatants carrying the shRNA expression construct KH1 containing either a scrambled or a sequence targeting p53 <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0022096#pone.0022096-Verhaegen1" target="_blank">[41]</a>. On Day 4 following infection total cell lysates were analysed for p53 expression by immunoblotting (lower part). Equal loading was controlled by Ponceau S staining of the blot. In the upper part the corresponding luciferase activity is depicted, normalized to the relative luciferase load of the cell lines determined by Real time PCR.</p

p53 expression in melanoma tissues.

<p>(<b>A</b>) Immunohistochemical detection of p53 in a primary melanoma. (<b>B</b>) 104 primary cutaneous and 81 metastatic melanoma samples were stained for p53. The percentage of positively stained tumor cells is given as bar graph.</p

Reproducibility of p53 sequencing.

<p>Sequencing chromatograms depicting the only reproducible p53 mutation and an example of an apparent mutation which could not be reproduced in an independent PCR amplification from the same sample.</p

Seroconversion with progress of disease.

<p>CD28 abs measured in one female patient during a period of more than 10 years in association with the course of the disease.</p

Specificity of the CD28 autoantibody Elisa.

<p>Human recombinant proteins were immobilized and measured by Elisa as described in the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0058087#s2" target="_blank">methods</a> part. Sera from 2 patients were used. No unspecific signal was detected. ACT-g: actin-γ, MAZ: myc-ass. zinc finger protein, PC9: pyruvate-carboxylase 9, SLP2: stomatin-like protein 2, ATG13: autophagy related 13 homolog.</p

Detailed data of melanoma patients.

<p>SSM = superficial spreading melanoma, NMM = nodular malignant melanoma, LMM = Lentigo maligna melanoma, ALM = acrolentiginous melanoma, AMM = amelanotic melanoma, UCM = unclassified melanoma, others = melanoma of the mucosa or uvea.</p

Data_Sheet_1.DOCX

<p>Persistent genus β-HPV (human papillomavirus) infection is a major co-factor for non-melanoma skin cancer in patients suffering from the inherited skin disease epidermodysplasia verruciformis (EV). Malignant EV lesions are particularly associated with HPV type 5 or 8. There is clinical and molecular evidence that HPV8 actively suppresses epithelial immunosurveillance by interfering with the recruitment of Langerhans cells, which may favor viral persistence. Mechanisms how persistent HPV8 infection promotes the carcinogenic process are, however, less well understood. In various tumor types chronic inflammation has a central role in tumor progression. The calprotectin complex consisting of S100A8 and S100A9 proteins has recently been identified as key driver of chronic and tumor promoting inflammation in skin carcinogenesis. It induces chemotaxis of neutrophil granulocytes and modulates inflammatory as well as immune responses. In this study, we demonstrate that skin lesions of EV-patients are massively infiltrated by inflammatory cells, including CD15⁺ granulocytes. At the same time we observed a very strong expression of S100A8 and S100A9 proteins in lesional keratinocytes, which was mostly confined to the suprabasal layers of the epidermis. Both proteins were hardly detected in non-lesional skin. Further experiments revealed that the HPV8 oncoproteins E6 and E7 were not involved in S100A8/A9 up-regulation. They rather suppressed differentiation-induced S100A8/A9 expression. In contrast, the viral transcription factor E2 strongly enhanced PMA-mediated S100A8/A9 up-regulation in primary human keratinocytes. Similarly, a tremendous up-regulation of both S100 proteins was observed, when minute amounts of the PMA-inducible CCAAT/enhancer binding protein β (C/EBPβ), which is expressed at low levels in the suprabasal layers of the epidermis, were co-expressed together with HPV8 E2. This confirmed our previous observation that C/EBPβ interacts and functionally synergizes with the HPV8 E2 protein in differentiation-dependent gene expression. Potent synergistic up-regulation of S100A8/A9 was seen at transcriptional and protein levels. S100A8/A9 containing supernatants from keratinocytes co-expressing HPV8 E2 and C/EBPβ significantly induced chemotaxis of granulocytes in migration assays supporting the relevance of our finding. In conclusion, our data suggest that the HPV8 E2 protein actively contributes to the recruitment of myeloid cells into EV skin lesions, which may support chronic inflammation and progression to skin cancer.</p

The inhibitory effect of human CD28 autoantibodoes is titer-dependent.

<p>Jurkat cells were incubated with sera derived from patient #64 that had been collected at different time points and that showed different titers of CD28 autoantibodies. Sera derived from a healthy donor (HD) or from a melanoma patients without CD28 autoantibodies were used as control. Cell viability was measured by EZ4U assay.</p