Publications Search Results
Now showing 1 - 10 of 18
PublicationCentral role for dermal fibroblasts in skin model protection against Candida albicans( 2017)
;Kühbacher, A. ;Henkel, H. ;Stevens, P. ;Grumaz, C. ;Finkelmeier, D. ;Burger-Kentischer, A. ;Sohn, K.Rupp, S.The fungal pathogen Candida albicans colonizes basically all human epithelial surfaces, including the skin. Under certain conditions, such as immunosuppression, invasion of the epithelia occurs. Not much is known about defense mechanisms against C. albicans in subepithelial layers such as the dermis. Using immune cell-supplemented 3D skin models we defined a new role for fibroblasts in the dermis and identified a minimal set of cell types for skin protection against C. albicans invasion. Dual RNA sequencing of individual host cell populations and C. albicans revealed that dermal invasion is directly impeded by dermal fibroblasts. They are able to integrate signals from the pathogen and CD4+ T cells and shift toward an antimicrobial phenotype with broad specificity that is dependent on Toll-like receptor 2 and interleukin 1Î². These results highlight a central function of dermal fibroblasts for skin protection, opening new possibilities for treatment of infectious diseases.
PublicationAn in vitro HSV-1 reactivation model containing quiescently infected PC12 cells( 2013)
;Hogk, I. ;Kaufmann, M. ;Finkelmeier, D. ;Rupp, S.Burger-Kentischer, A.Advances in the understanding of the infection and reactivation process of herpes simplex type 1 (HSV-1) are generally gained by monolayer cultures or extensive and cost-intensive animal models. So far, no reliable in vitro skin model exists either to investigate the molecular mechanisms involved in controlling latency and virus reactivation or to test pharmaceuticals. Here we demonstrate the first in vitro HSV-1 reactivation model generated by using the human keratinocyte cell line HaCaT grown on a collagen substrate containing primary human fibroblasts. We integrated the unique feature of a quiescently infected neuronal cell line, the rat pheochromocytoma line PC12, within the dermal layer of the three-dimensional skin equivalent. Transmission electron microscopy, a cell-based TCID50 assay, and polymerase chain reaction analysis were used to verify cell latency. Thereby viral DNA could be detected, whereas extracellular as well as intracellular virus activity could not be found. Further, the infected PC12 cells show no spontaneous reactivation within the in vitro skin equivalent. In order to simulate a physiologically comparable HSV-1 infection, we achieved a specific and pointed reactivation of quiescently HSV-1 infected PC12 cells by UVB irradiation at 1000 mJ/cm2.
PublicationIdentification and characterisation of novel antifungal compounds against fungal human pathogens( 2012)
;Keller, P.D. ;Burger-Kentischer, A. ;Finkelmeier, D. ;Wiesmuller, K.H. ;Lemuth, K. ;Hiller, E. ;Engelhardt, I. ;Muller, C. ;Schroppel, K. ;Bracher, F.Rupp, S.
PatentIn vitro- Testsystem für virale Infektionen( 2011)
;Burger-Kentischer, A. ;Hogk, I. ;Finkelmeier, D. ;Walles, H. ;Rupp, S.Kaufmann, M.Multilayered biological in vitro tissue comprising a dermis layer containing a collagen biomatrix with fibroblasts embedded in it, and an epidermis layer containing epithelial cells, is claimed, where latently virally infected neuronal cells are integrated into the dermis layer. Independent claims are included for: (1) a method for producing the multilayered biological tissue as an in vitro test system, comprising providing a multilayered tissue containing a dermis layer, a collagen biomatrix with fibroblasts embedded in it, and an epidermis layer containing epithelial cells, and seeding virally infected neuronal cells on the multilayered biological tissue so that they are integrated into the biological tissue; and (2) a method for finding an active ingredient with an antiviral action from a group of substances to be examined, comprising providing the multilayered biological in vitro tissue, reacting the virus infection in the latently virally infected neuronal cells of the tissue, bringing the tissue before or after the virus activation into contact with the substance to be examined, and verifying the extent of the virus activation, where the reduction of the extent of the virus activation compared to tissue not brought into contact with the substance indicates an antiviral action of the substance.
PublicationA new class of antimycotic (S)-2-aminoalkyl benzimidazoles exhibits potent antifungal activity against clinically relevant and against fluconazole resistant strains of Candida spp( 2011)
;Bauer, J. ;Kinast, S. ;Burger-Kentischer, A. ;Finkelmeier, D. ;Kleymann, G. ;Abu Rayyan, W. ;Schroppel, K. ;Singh, A. ;Spohn, R. ;Jung, G. ;Wiesmüller, K.-H. ;Rupp, S.Eickhoff, H.
PublicationIdentification and characterisation of novel antifungal compounds using a screening assay based on host-pathogen interaction models( 2011)
;Keller, P. ;Burger-Kentischer, A. ;Finkelmeier, D. ;Kleymann, G. ;Wiesmuller, K.H. ;Lemuth, K. ;Hiller, E.Rupp, S.
PublicationA screening assay based on host-pathogen interaction models identifies a set of novel antifungal benzimidazole derivatives( 2011)
;Burger-Kentischer, A. ;Finkelmeier, D. ;Keller, P. ;Bauer, J. ;Eickhoff, H. ;Kleymann, G. ;Rayyan, W.A. ;Singh, A. ;Schröppel, K. ;Lemuth, K. ;Wiesmüller, K.-H.Rupp, S.Fungal infections are a serious health problem in clinics, especially in the immune-compromised patient. Disease ranges from widespread superficial infections like vulvovaginal infections to life-threatening systemic candidiasis. Especially for systemic mycoses, only a limited arsenal of antifungals is available. The most commonly used classes of antifungal compounds used include azoles, polyenes, and echinocandins. Due to emerging resistance to standard therapy, significant side effects, and high costs for several antifungals, there is a medical need for new antifungals in the clinic and general practice. In order to expand the arsenal of compounds with antifungal activities, we screened a compound library including more than 35,000 individual compounds derived from organic synthesis as well as combinatorial compound collections representing mixtures of compounds for antimycotic activity. In total, more than 100,000 compounds were screened using a new type of activity- selectivity assay, analyzing both the antifungal activity and the compatibility with human cells at the same time. One promising hit, an (S)-2-aminoalkyl benzimidazole derivative, was developed among a series of lead compounds showing potent antifungal activity. (S)-2-(1-Aminoisobutyl)-1-(3- chlorobenzyl) benzimidazole showed the highest antifungal activity and the best compatibility with human cells in several cell culture models and against a number of clinical isolates of several species of pathogenic Candida yeasts. Transcriptional profiling indicates that the newly discovered compound is a potential inhibitor of the ergosterol pathway, in contrast to other benzimidazole derivatives, which target microtubules.
PublicationHigh-throughput-screening-based identification and structure-activity relationship characterization defined (S)-2-(1-aminoisobutyl)-1-(3-chlorobenzyl) benzimidazole as a highly antimycotic agent nontoxic to cell lines( 2011)
;Bauer, J. ;Kinast, S. ;Burger-Kentischer, A. ;Finkelmeier, D. ;Kleymann, G. ;Rayyan, W.A. ;Schröppel, K. ;Singh, A. ;Jung, G. ;Wiesmüller, K.-H. ;Rupp, S.Eickhoff, H.Novel nontoxic (S)-2-aminoalkylbenzimidazole derivatives were found to be effective against Candida spp. at low micromolar concentrations using high-throughput screening with infected HeLa cells. A collection of analogues defined the chemical groups relevant for activity. The most active compound was characterized by transcriptional analysis of the response of C. albicans Sc5314. (S)-2-(1-Aminoisobutyl)-1-(3-chlorobenzyl)benzimidazole had a strong impact on membrane biosynthesis. Testing different clinically relevant pathogenic fungi showed the selectivity of the antimycotic activity against Candida species.
PublicationA new cell-based innate immune receptor assay for the examination of receptor activity, ligand specificity, signalling pathways and the detection of pyrogens( 2010)
;Burger-Kentischer, A. ;Abele, I.S. ;Finkelmeier, D. ;Wiesmüller, K.-H.Rupp, S.The pattern recognition receptors (PRRs) of the innate immune system are the first defence line of the immune system. Toll-like receptors (TLRs) are the most well known and the best examined of the PR receptors. In the last years TLRs had been studied in different ways resulting in a lot of new insights in the function and signalling pathways of these receptors. However, it was not possible to investigate individual combinations of the TLRs and their specific ligands, because of the complex network in immune signalling resulting in interference with each other. This work shows a new cell-based assay, established for the analysis of single PRRs or heterodimers. For this purpose NIH3T3 (mouse fibroblasts) were stably transfected with the NF-?B-inducible reporter gene secreted alkaline phosphatase (SEAP) together with the corresponding combinations of human TLRs and their co-receptors (e.g. TLR1/2, TLR2/6 and TLR4/CD14). The specificity of the respective cell lines was shown by induction with variations of specific and unspecific ligands (immune-stimulating components of microorganisms or synthetic ligands). Analysis via the NF-Kappa B-dependent reporter gene SEAP allows a direct way to detect the human TLR-activity. Our results showed that this assay is highly sensitive and specific for the respective ligands. For the synthetic ligands Pam2CysSK4 the assay demonstrates a detection limit of 1 pg/ml for TLR2/6. In summary, this test system allows the investigation of individual human PRR-receptors in a highly specific way, without interference with other immune components opening new avenues for novel insights in the innate immune system and its applications.