A hámsejtek immunológiai működésében résztvevő gének azonosítása DNS-microarray módszerrel = Identification of genes involved in the immune-function of epithelial cells using DNA microarray.

A hámsejtek immunológiai működésében résztvevő gének azonosítása DNS-microarray módszerrel Az emberi szervezetet a külső környezettől elválasztó legfontosabb határoló szervünk a bőr, amely egyrészt mechanikai védelmet nyújt a külvilág káros hatásai ellen, másrészt aktív, protektív feladatokat is ellát. Munkacsoportunk a bőrnek és a hámsejteknek a veleszületett immunitásban betöltött szerepét kutatja. Vizsgálataink során cDNS microarray módszerrel vizsgáltuk azt a kérdést, hogy a különféle bakteriális és gomba eredetű anyagok (LPS, PGN, hővel elölt Candida albicans, élő Propionibacterium acnes) milyen génexpresszós változásokat okoznak primer tenyésztett keratinocitákban. Eredményeink alapján igazoltuk, hogy a keratinocitákban gyulladásos citokin, kemokin, és antimikrobiális hatású fehérjét kódoló gének kifejeződése megváltozik a patogén eredetű anyagok hatására. A folyamatok szabályozásában a Toll-like receptorok, valamint NF-kB játszik központi szerepet. Igazoltuk, hogy a patogének támadásának kitett testfelületeken számos nem immun-eredetű sejttípus rendelkezik a kórokozók felismerésének képességével, ezek bakteriális és gomba-eredetű anyagok hatására képesek a veleszületett és adaptív immunfolyamatok elindítására és szabályozására. Eredményeink elméleti jelentősége mellett egy olyan tesztrendszer bevezetését is lehetővé tették, mely alkalmas gyulladásos bőrbetegségek esetén alkalmazott lokális immunmoduláló szerek hatásmechanizmusának vizsgálatára. | Identification of genes involved in the immune-function of epithelial cells using DNA microarray. The human skin is the most important barrier of the organism. It can not only passively separate the outer environment from the inner body, but also plays an active protective function. Our main focus is the better understanding of the innate immune functions of epidermal keratinocytes. For this reason we have investigated the global gene expression changes in response to various microbial components (PLS, PGN, heat-killed Candida albicans, and live Propionibacterium acnes) in primary keratinocytes using the microarray technique. Based on our results we could show, that the expression of various pro-inflammatory cytokine, chemokine, and antimicrobial peptide coding genes are changing in response to pathogen derived chemical compounds, and whole pathogen cells. Toll-like receptors and NF-kB are playing an important role in the regulation of these genes. Our results also suggest, that this ability of the keratinocytes is not unique to them, other cell types on various surfaces of the body where pathogen attack happens regularly can also recognize the microbes and initiate and regulate immune processes. Based on our data, we also set up an experimental model system that can be used for the evaluation of the exact function of various topical immunmodulatory chemicals used to treat inflammatory skin diseases

Distinct Strains of Propionibacterium acnes Induce Selective Human β-Defensin-2 and Interleukin-8 Expression in Human Keratinocytes Through Toll-Like Receptors

Acne is a chronic inflammatory disease of the pilosebaceous follicle. One of the main pathogenetic factors in acne is the increased proliferation of Propionibacterium acnes (P. acnes) in the pilosebaceous unit. We investigated whether direct interaction of P. acnes with keratinocytes might be involved in the inflammation and ductal hypercornification in acne. The capacities of different P. acnes strains to activate the innate immune response and the growth of cultured keratinocytes were investigated. We have found that two clinical isolates of P. acnes significantly induced human β-defensin-2 (hBD2) messenger RNA (mRNA) expression; in contrast a third clinical isolate and the reference strain (ATCC11828) had no effect on hBD2 mRNA expression. In contrast, all four strains significantly induced the interleukin-8 (IL-8) mRNA expression. The P. acnes-induced increase in hBD2 and IL-8 gene expression could be inhibited by anti-Toll-like receptor 2 (TLR2) and anti-TLR4 neutralizing antibodies, suggesting that P. acnes-induced secretion of soluble factors in keratinocytes are both TLR2 and TLR4 dependent. In addition, the clinical isolate P. acnes (889) was capable of inducing keratinocyte cell growth in vitro. Our findings suggest that P. acnes modulates the antimicrobial peptide and chemokine expression of keratinocytes and thereby contributes to the recruitment of inflammatory cells to the sites of infections

Benignus és malignus hiperproliferatív ill. gyulladásos bőrbetegségek pathomechanizmusának vizsgálata funkcionális genomikai módszerekkel = Studies on the pathomechanism of benign and malign hyperproliferative as well as inflammatory skin diseases by means of functional genomics

Pikkelysömörben vizsgáltuk a sejtciklus szabályozásban részt vevő extracelluláris mátrix protein, a fibronektin szerepét, és azonosítottunk egy nem kódoló RNS gént (PRINS), mely szerepet játszik a pikkelysömörre való hajlam, ill. a sejtek stressz válaszának kialakításában. A multifaktoriális etiológiájú lábszárfekély genetikai hátterének vizsgálata során azonosítottunk egy SNP-t, amely magasabb arányban fordul elő a betegekben, mint egészségesekben, feltételezzük, hogy szerepet játszik a betegség pathogenezisében. A serdülőkorban és gyakran még a felnőttkorban is gondot jelentő acne vulgaris gyulladásos bőrbetegség genomikai kutatása során igazoltuk, hogy az interleukin 1 receptor antagonista gén (IL-1RN) egy polimorfizmusa ebben a kórképben magasabb arányban fordul elő, mint az egészséges populációban, amely pathogenetikai szerepére utal. Megmutattuk, hogy az acne pathomechanizmusában kulcsfontosságú Propionibacterium acnes baktérium a keratinocitákban a veleszületett immunitásban szerepet játszó gének expresszióját indukálja. Vizsgáltuk a hisztamin szerepét az allergiás kontakt dermatitisz pathogenezisében. Kísérleteinkhez hisztidin dekarboxiláz knock-out egereket használtunk, és eredményeink alapján feltételezzük, hogy a hisztamin a késői típusú kontakt dermatitiszben negatív reguláló szerepet tölt be. A pályázat keretében a gyulladásos bőrbetegségek ill. a szénanátha kezelésére alkalmas új fototerápiás eljárásokat is fejlesztettünk és vizsgáltuk azok hatásmechanizmusát. | We investigated the role of an extracellular matrix protein, fibronectin (an important regulator of the cell cycle), in psoriasis. We identified a non-coding RNA gene, PRINS playing a role in cell stress, and a key factor of psoriasis susceptibility. The genetic background of the multifactorial leg ulcer was also studied. We discovered an SNP that is more frequent among leg ulcer patients than in controls. This finding suggests that this SNP might play a role in the pathogenesis of this disease. Acne vulgaris is another very common inflammatory skin disorder affecting a significant proportion of the teenager and the adult population. We showed that a polymorphism of the interleukin 1 receptor antagonist gene (IL1-RN) is more frequent among the affected individuals compared to healthy controls. This suggests a role for the IL1-RN gene in the development of this skin condition. Studies on the contribution of the skin colonizing Propionibacterium acne in the pathomechanism of acne showed that this bacterium induces the expression of genes playing a role in innate immunity of human kerationocytes. The role of histamine in allergic contact dermatitis was also investigated. We used histamine decarboxylase knock-out mice, and our results suggest that histamine plays a negative regulatory role in the late-type contact dermatitis. We have developed novel phototherapeutic methods for the treatment of inflammatory skin diseases and rhinitis allergica, and studied their mode of action

MicroRNAs: Novel Regulators Involved in the Pathogenesis of Psoriasis?

MicroRNAs are a recently discovered class of posttranscriptional regulators of gene expression with critical functions in health and disease. Psoriasis is the most prevalent chronic inflammatory skin disease in adults, with a substantial negative impact on the patients' quality of life. Here we show for the first time that psoriasis-affected skin has a specific microRNA expression profile when compared with healthy human skin or with another chronic inflammatory skin disease, atopic eczema. Among the psoriasis-specific microRNAs, we identified leukocyte-derived microRNAs and one keratinocyte-derived microRNA, miR-203. In a panel of 21 different human organs and tissues, miR-203 showed a highly skin-specific expression profile. Among the cellular constituents of the skin, it was exclusively expressed by keratinocytes. The up-regulation of miR-203 in psoriatic plaques was concurrent with the down-regulation of an evolutionary conserved target of miR-203, suppressor of cytokine signaling 3 (SOCS-3), which is involved in inflammatory responses and keratinocyte functions. Our results suggest that microRNA deregulation is involved in the pathogenesis of psoriasis and contributes to the dysfunction of the cross talk between resident and infiltrating cells. Taken together, a new layer of regulatory mechanisms is involved in the pathogenesis of chronic inflammatory skin diseases

A Mannose-Binding Receptor is Expressed on Human Keratinocytes and Mediates Killing of Candida albicans

Human keratinocytes are known to kill Candida albicans in vitro, but the mechanism of killing is not yet understood. Here, we demonstrate that spontaneous, ultraviolet-B-light-induced, α-melanocyte-stimulating-hormone-induced, and interleukin-8-induced Candida killing by keratinocytes can be inhibited with mannan and mannosylated bovine serum albumin (Man-BSA). A polyclonal goat serum raised against the human macrophage mannose receptor stained suprabasal keratinocytes, but no staining was observed on keratinocytes with a monoclonal antibody (mAb15) specific for the human macrophage mannose receptor. Mannose-affinity chromatography of keratinocyte extract isolated a 200 kDa protein, and on the Western blot the goat antiserum reacted with a 200 kDa protein. In radioligand binding studies, the binding of 125I-Man-BSA to human keratinocytes was inhibited by mannan in a concentration-dependent manner. Analysis of the binding revealed a single class keratinocyte mannose receptor with a KD of 1.4 × 10-8 M and a Bmax of 1 × 104 binding sites per cell. The binding of 125I-Man-BSA to keratinocytes proved to be time-dependent, acid-precipitable, and Ca2+- and trypsin-sensitive. After trypsinization the receptors underwent a rapid recovery at 37°C. These results demonstrate the presence of mannose receptor on human keratinocytes, and its active involvement in the killing of Candida albicans

Histidine Decarboxylase Expression in Human Melanoma

SummaryHistamine has been implicated as one of the mediators involved in regulation of proliferation in both normal and neoplastic tissues. Histidine decarboxylase, the only enzyme that catalyzes the formation of histamine from L-histidine, is an essential regulator of histamine levels. In this study, we investigated the gene and protein expression of histidine decarboxylase in melanoma. Reverse transcriptase polymerase chain reaction and in situ hybridization studies of WM-35, WM-983/B, HT-168, and M1 human melanoma cell lines both resulted in positive signals for histidine decarboxylase messenger RNA. A polyclonal chicken antibody was developed against human histidine decarboxylase and protein expression was confirmed by western blot analysis of the cell lysates, revealing a predominant immunoreactive band at approximately 54 kDa corresponding to monomeric histidine decarboxylase. Protein expression of histidine decarboxylase was also shown by flow cytometric analysis and strong punctate cytoplasmic staining of melanoma cell lines. Moreover, both primary and metastatic human melanoma tissues were brightly stained for histidine decarboxylase. When compared with the very weak or no reactions on cultivated human melanocytes both western blot and immunohistochemical studies showed much stronger histidine decarboxylase expression in melanoma cells. These findings suggest that expression of histidine decarboxylase is elevated in human melanoma

Correction to: EGFR/Ras-induced CCL20 production modulates the tumour microenvironment

The article ‘EGFR/Ras-induced CCL20 production modulates the tumour microenvironment’, written by Andreas Hippe, Stephan Alexander Braun, Péter Oláh, Peter Arne Gerber, Anne Schorr, Stephan Seeliger, Stephanie Holtz, Katharina Jannasch, Andor Pivarcsi, Bettina Buhren, Holger Schrumpf, Andreas Kislat, Erich Bünemann, Martin Steinhoff, Jens Fischer, Sérgio A. Lira, Petra Boukamp, Peter Hevezi, Nikolas Hendrik Stoecklein, Thomas Hoffmann, Frauke Alves, Jonathan Sleeman, Thomas Bauer, Jörg Klufa, Nicole Amberg, Maria Sibilia, Albert Zlotnik, Anja Müller- Homey and Bernhard Homey, was originally published electronically on the publisher’s internet portal on 30 June 2020 without open access. With the author(s)’ decision to opt for Open Choice the copyright of the article changed on 16 September 2021 to © The Author(s) 2021 and the article is forthwith distributed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/ licenses/by/4.0/. Open Access funding enabled and organized by Projekt DEAL

Human antimicrobial protein hCAP18/LL-37 promotes a metastatic phenotype in breast cancer

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