TY  - JOUR
A1  - Bröker, Katharine
A1  - Sinelnikov, Evgeny
A1  - Gustavus, Dirk
A1  - Schumacher, Udo
A1  - Pörtner, Ralf
A1  - Hoffmeister, Hans
A1  - Lüth, Stefan
A1  - Dammermann, Werner
T1  - Mass Production of Highly Active NK Cells for Cancer Immunotherapy in a GMP Conform Perfusion Bioreactor
JF  - Frontiers in Bioengineering and Biotechnology
N2  - NK cells have emerged as promising candidates for cancer immunotherapy, especially due to their ability to fight circulating tumor cells thereby preventing metastases formation. Hence several studies have been performed to generate and expand highly cytotoxic NK cells ex vivo, e.g., by using specific cytokines to upregulate both their proliferation and surface expression of distinct activating receptors. Apart from an enhanced activity, application of NK cells as immunotherapeutic agent further requires sufficient cell numbers and a high purity. All these parameters depend on a variety of different factors including the starting material, additives like cytokines as well as the culture system. Here we analyzed PBMC-derived NK cells of five anonymized healthy donors expanded under specific conditions in an innovative perfusion bioreactor system with respect to their phenotype, IFN gamma production, and cytotoxicity in vitro. Important features of the meander type bioreactors used here are a directed laminar flow of medium and control of relevant process parameters. Cells are cultivated under "steady state" conditions in perfusion mode. Our data demonstrate that expansion of CD3(+) T cell depleted PBMCs in our standardized system generates massive amounts of highly pure (>85%) and potent anticancer active NK cells. These cells express a variety of important receptors driving NK cell recruitment, adhesion as well as activation. More specifically, they express the chemokine receptors CXCR3, CXCR4, and CCR7, the adhesion molecules L-selectin, LFA-1, and VLA-4, the activating receptors NKp30, NKp44, NKp46, NKG2D, DNAM1, and CD16 as well as the death ligands TRAIL and Fas-L. Moreover, the generated NK cells show a strong IFN gamma expression upon cultivation with K562 tumor cells and demonstrate a high cytotoxicity toward leukemic as well as solid tumor cell lines in vitro. Altogether, these characteristics promise a high clinical potency of thus produced NK cells awaiting further evaluation.
KW  - natural killer cells (NK cells)
KW  - cytotoxicity
KW  - tumor immunity
KW  - immunotherapy
KW  - perfusion bioreactor
KW  - GMP
KW  - mass production process
Y1  - 2019
U6  - https://doi.org/10.3389/fbioe.2019.00194
SN  - 2296-4185
VL  - 7
PB  - Frontiers Research Foundation
CY  - Lausanne
ER  - 
TY  - JOUR
A1  - Zalden, Peter
A1  - Quirin, Florian
A1  - Schumacher, Mathias
A1  - Siegel, Jan
A1  - Wei, Shuai
A1  - Koc, Azize
A1  - Nicoul, Matthieu
A1  - Trigo, Mariano
A1  - Andreasson, Pererik
A1  - Enquist, Henrik
A1  - Shu, Michael J.
A1  - Pardini, Tommaso
A1  - Chollet, Matthieu
A1  - Zhu, Diling
A1  - Lemke, Henrik
A1  - Ronneberger, Ider
A1  - Larsson, Jörgen
A1  - Lindenberg, Aaron M.
A1  - Fischer, Henry E.
A1  - Hau-Riege, Stefan
A1  - Reis, David A.
A1  - Mazzarello, Riccardo
A1  - Wuttig, Matthias
A1  - Sokolowski-Tinten, Klaus
T1  - Femtosecond x-ray diffraction reveals a liquid-liquid phase transition in phase-change materials
JF  - Science
N2  - In phase-change memory devices, a material is cycled between glassy and crystalline states. The highly temperature-dependent kinetics of its crystallization process enables application in memory technology, but the transition has not been resolved on an atomic scale. Using femtosecond x-ray diffraction and ab initio computer simulations, we determined the time-dependent pair-correlation function of phase-change materials throughout the melt-quenching and crystallization process. We found a liquid-liquid phase transition in the phase-change materials Ag4In3Sb67Te26 and Ge15Sb85 at 660 and 610 kelvin, respectively. The transition is predominantly caused by the onset of Peierls distortions, the amplitude of which correlates with an increase of the apparent activation energy of diffusivity. This reveals a relationship between atomic structure and kinetics, enabling a systematic optimization of the memory-switching kinetics.
Y1  - 2019
U6  - https://doi.org/10.1126/science.aaw1773
SN  - 0036-8075
SN  - 1095-9203
VL  - 364
IS  - 6445
SP  - 1062
EP  - 1067
PB  - American Assoc. for the Advancement of Science
CY  - Washington, DC
ER  - 
TY  - JOUR
A1  - Scheller, Frieder W.
A1  - Yarman, Aysu
A1  - Bachmann, Till
A1  - Hirsch, Thomas
A1  - Kubick, Stefan
A1  - Renneberg, Reinhard
A1  - Schumacher, Soeren
A1  - Wollenberger, Ursula
A1  - Teller, Carsten
A1  - Bier, Frank Fabian
ED  - Gu, MB
ED  - Kim, HS
T1  - Future of biosensors: a personal view
JF  - Advances in biochemical engineering, biotechnology
JF  - Advances in Biochemical Engineering-Biotechnology
N2  - Biosensors representing the technological counterpart of living senses have found routine application in amperometric enzyme electrodes for decentralized blood glucose measurement, interaction analysis by surface plasmon resonance in drug development, and to some extent DNA chips for expression analysis and enzyme polymorphisms. These technologies have already reached a highly advanced level and need minor improvement at most. The dream of the "100-dollar' personal genome may come true in the next few years provided that the technological hurdles of nanopore technology or of polymerase-based single molecule sequencing can be overcome. Tailor-made recognition elements for biosensors including membrane-bound enzymes and receptors will be prepared by cell-free protein synthesis. As alternatives for biological recognition elements, molecularly imprinted polymers (MIPs) have been created. They have the potential to substitute antibodies in biosensors and biochips for the measurement of low-molecular-weight substances, proteins, viruses, and living cells. They are more stable than proteins and can be produced in large amounts by chemical synthesis. Integration of nanomaterials, especially of graphene, could lead to new miniaturized biosensors with high sensitivity and ultrafast response. In the future individual therapy will include genetic profiling of isoenzymes and polymorphic forms of drug-metabolizing enzymes especially of the cytochrome P450 family. For defining the pharmacokinetics including the clearance of a given genotype enzyme electrodes will be a useful tool. For decentralized online patient control or the integration into everyday "consumables' such as drinking water, foods, hygienic articles, clothing, or for control of air conditioners in buildings and cars and swimming pools, a new generation of "autonomous' biosensors will emerge.
KW  - Biosensors
KW  - Molecularly imprinted polymers
KW  - Personalized medicine
Y1  - 2014
SN  - 978-3-642-54143-8; 978-3-642-54142-1
U6  - https://doi.org/10.1007/10_2013_251
SN  - 0724-6145
VL  - 140
SP  - 1
EP  - 28
PB  - Springer
CY  - Berlin
ER  - 
TY  - JOUR
A1  - Schumacher-Wandersleb, Michael H. M. G.
A1  - Petersen, Stefan
A1  - Peter, Martin G.
T1  - Preparation of the N-Acetylglucosaminidase inhibitor 1-Acetamido-1,2,5-tride oxy-2,5-imino-D-glucitol from methyl a-D-Mannopyranoside
Y1  - 1994
ER  - 
TY  - JOUR
A1  - Peter, Martin G.
A1  - Ley, J. P.
A1  - Petersen, Stefan
A1  - Londershausen, M.
A1  - Schumacher-Wandersleb, Michael H. M. G.
A1  - Spindler, Klaus-Dieter
A1  - Spindler-Barth, Margarethe
A1  - Turberg, Andreas
T1  - Synthesis of chitinase inhibitors
Y1  - 1994
ER  - 
TY  - JOUR
A1  - Dejonghe, Wim
A1  - Kuenen, Sabine
A1  - Mylle, Evelien
A1  - Vasileva, Mina
A1  - Keech, Olivier
A1  - Viotti, Corrado
A1  - Swerts, Jef
A1  - Fendrych, Matyas
A1  - Ortiz-Morea, Fausto Andres
A1  - Mishev, Kiril
A1  - Delang, Simon
A1  - Scholl, Stefan
A1  - Zarza, Xavier
A1  - Heilmann, Mareike
A1  - Kourelis, Jiorgos
A1  - Kasprowicz, Jaroslaw
A1  - Nguyen, Le Son Long
A1  - Drozdzecki, Andrzej
A1  - Van Houtte, Isabelle
A1  - Szatmari, Anna-Maria
A1  - Majda, Mateusz
A1  - Baisa, Gary
A1  - Bednarek, Sebastian York
A1  - Robert, Stephanie
A1  - Audenaert, Dominique
A1  - Testerink, Christa
A1  - Munnik, Teun
A1  - Van Damme, Daniel
A1  - Heilmann, Ingo
A1  - Schumacher, Karin
A1  - Winne, Johan
A1  - Friml, Jiri
A1  - Verstreken, Patrik
A1  - Russinova, Eugenia
T1  - Mitochondrial uncouplers inhibit clathrin-mediated endocytosis largely through cytoplasmic acidification
JF  - Nature Communications
N2  - ATP production requires the establishment of an electrochemical proton gradient across the inner mitochondrial membrane. Mitochondrial uncouplers dissipate this proton gradient and disrupt numerous cellular processes, including vesicular trafficking, mainly through energy depletion. Here we show that Endosidin9 (ES9), a novel mitochondrial uncoupler, is a potent inhibitor of clathrin-mediated endocytosis (CME) in different systems and that ES9 induces inhibition of CME not because of its effect on cellular ATP, but rather due to its protonophore activity that leads to cytoplasm acidification. We show that the known tyrosine kinase inhibitor tyrphostinA23, which is routinely used to block CME, displays similar properties, thus questioning its use as a specific inhibitor of cargo recognition by the AP-2 adaptor complex via tyrosine motif-based endocytosis signals. Furthermore, we show that cytoplasm acidification dramatically affects the dynamics and recruitment of clathrin and associated adaptors, and leads to reduction of phosphatidylinositol 4,5-biphosphate from the plasma membrane.
Y1  - 2016
U6  - https://doi.org/10.1038/ncomms11710
SN  - 2041-1723
VL  - 7
SP  - 1959
EP  - 1968
PB  - Nature Publ. Group
CY  - London
ER  - 
TY  - JOUR
A1  - Giulbudagian, Michael
A1  - Hönzke, Stefan
A1  - Bergueiro, Julián
A1  - Işık, Doğuş
A1  - Schumacher, Fabian
A1  - Saeidpour, Siavash
A1  - Lohan, Silke
A1  - Meinke, Martina
A1  - Teutloff, Christian
A1  - Schäfer-Korting, Monika
A1  - Yealland, Guy
A1  - Kleuser, Burkhard
A1  - Hedtrich, Sarah
A1  - Calderón, Marcelo
T1  - Enhanced topical delivery of dexamethasone by beta-cyclodextrin decorated thermoresponsive nanogels
JF  - Nanoscale
N2  - Highly hydrophilic, responsive nanogels are attractive as potential systems for the topical delivery of bioactives encapsulated in their three-dimensional polymeric scaffold. Yet, these drug carrier systems suffer from drawbacks for efficient delivery of hydrophobic drugs. Addressing this, β-cyclodextrin (βCD) could be successfully introduced into the drug carrier systems by exploiting its unique affinity toward dexamethasone (DXM) as well as its role as topical penetration enhancer. The properties of βCD could be combined with those of thermoresponsive nanogels (tNGs) based on dendritic polyglycerol (dPG) as a crosslinker and linear thermoresponsive polyglycerol (tPG) inducing responsiveness to temperature changes. Electron paramagnetic resonance (EPR) studies localized the drug within the hydrophobic cavity of βCD by differences in its mobility and environmental polarity. In fact, the fabricated carriers combining a particulate delivery system with a conventional penetration enhancer, resulted in an efficient delivery of DXM to the epidermis and the dermis of human skin ex vivo (enhancement compared to commercial DXM cream: ∼2.5 fold in epidermis, ∼30 fold in dermis). Furthermore, DXM encapsulated in βCD tNGs applied to skin equivalents downregulated the expression of proinflammatory thymic stromal lymphopoietin (TSLP) and outperformed a commercially available DXM cream.
Y1  - 2017
U6  - https://doi.org/10.1039/c7nr04480a
SN  - 2040-3364
SN  - 2040-3372
VL  - 10
IS  - 1
SP  - 469
EP  - 479
PB  - Royal Society of Chemistry
CY  - Cambridge
ER  - 
TY  - JOUR
A1  - Döge, Nadine
A1  - Hönzke, Stefan
A1  - Schumacher, Fabian
A1  - Balzus, Benjamin
A1  - Colombo, Miriam
A1  - Hadam, Sabrina
A1  - Rancan, Fiorenza
A1  - Blume-Peytavi, Ulrike
A1  - Schäfer-Korting, Monika
A1  - Schindler, Anke
A1  - Rühl, Eckart
A1  - Skov, Per Stahl
A1  - Church, Martin K.
A1  - Hedtrich, Sarah
A1  - Kleuser, Burkhard
A1  - Bodmeier, Roland
A1  - Vogt, Annika
T1  - Ethyl cellulose nanocarriers and nanocrystals differentially deliver dexamethasone into intact, tape-stripped or sodium lauryl sulfate-exposed ex vivo human skin - assessment by intradermal microdialysis and extraction from the different skin layers
JF  - Journal of controlled release
N2  - Understanding penetration not only in intact, but also in lesional skin with impaired skin barrier function is important, in order to explore the surplus value of nanoparticle-based drug delivery for anti-inflammatory dermatotherapy. Herein, short-termex vivo cultures of (i) intact human skin, (ii) skin pretreated with tape-strippings and (iii) skin pre-exposed to sodium lauryl sulfate (SLS) were used to assess the penetration of dexamethasone (Dex). Intradermal microdialysis was utilized for up to 24 h after drug application as commercial cream, nanocrystals or ethyl cellulose nanocarriers applied at the therapeutic concentration of 0.05%, respectively. In addition, Dex was assessed in culture media and extracts from stratum corneum, epidermis and dermis after 24 h, and the results were compared to those in heat-separated split skin from studies in Franz diffusion cells. Providing fast drug release, nanocrystals significantly accelerated the penetration of Dex. In contrast to the application of cream and ethyl cellulose nanocarriers, Dex was already detectable in eluates after 6 h when applying nanocrystals on intact skin. Disruption of the skin barrier further accelerated and enhanced the penetration. Encapsulation in ethyl cellulose nanocarriers delayed Dex penetration. Interestingly, for all formulations highly increased concentrations in the dialysate were observed in tape-stripped skin, whereas the extent of enhancement was less in SLS-exposed skin. The results were confirmed in tissue extracts and were in line with the predictions made by in vitro release studies and ex vivo Franz diffusion cell experiments. The use of 45 kDa probes further enabled the collection of inflammatory cytokines. However, the estimation of glucocorticoid efficacy by Interleukin (IL)-6 and IL-8 analysis was limited due to the trauma induced by the probe insertion. Ex vivo intradermal microdialysis combined with culture media analysis provides an effective, skin-sparing method for preclinical assessment of novel drug delivery systems at therapeutic doses in models of diseased skin. (C) 2016 Elsevier B.V. All rights reserved.
KW  - Drug delivery systems
KW  - Polymeric nanoparticles
KW  - Dexamethasone
KW  - Microdialysis
KW  - Skin penetration
KW  - Skin barrier disruption
Y1  - 2016
U6  - https://doi.org/10.1016/j.jconrel.2016.07.009
SN  - 0168-3659
SN  - 1873-4995
VL  - 242
SP  - 25
EP  - 34
PB  - Elsevier
CY  - Amsterdam
ER  - 
TY  - JOUR
A1  - Gutbier, Birgitt
A1  - Schönrock, Stefanie M.
A1  - Ehrler, Carolin
A1  - Haberberger, Rainer
A1  - Dietert, Kristina
A1  - Gruber, Achim D.
A1  - Kummer, Wolfgang
A1  - Michalick, Laura
A1  - Kuebler, Wolfgang M.
A1  - Hocke, Andreas C.
A1  - Szymanski, Kolja
A1  - Letsiou, Eleftheria
A1  - Lüth, Anja
A1  - Schumacher, Fabian
A1  - Kleuser, Burkhard
A1  - Mitchell, Timothy J.
A1  - Bertrams, Wilhelm
A1  - Schmeck, Bernd
A1  - Treue, Denise
A1  - Klauschen, Frederick
A1  - Bauer, Torsten T.
A1  - Tönnies, Mario
A1  - Weissmann, Norbert
A1  - Hippenstiel, Stefan
A1  - Suttorp, Norbert
A1  - Witzenrath, Martin
T1  - Sphingosine Kinase 1 Regulates Inflammation and Contributes to Acute Lung Injury in Pneumococcal Pneumonia via the Sphingosine-1-Phosphate Receptor 2
JF  - Critical care medicine
N2  - Objectives: Severe pneumonia may evoke acute lung injury, and sphingosine-1-phosphate is involved in the regulation of vascular permeability and immune responses. However, the role of sphingosine-1-phosphate and the sphingosine-1-phosphate producing sphingosine kinase 1 in pneumonia remains elusive. We examined the role of the sphingosine-1-phosphate system in regulating pulmonary vascular barrier function in bacterial pneumonia. Design: Controlled, in vitro, ex vivo, and in vivo laboratory study. Subjects: Female wild-type and SphK1-deficient mice, 8-10 weeks old. Human postmortem lung tissue, human blood-derived macrophages, and pulmonary microvascular endothelial cells. Interventions: Wild-type and SphK1-deficient mice were infected with Streptococcus pneumoniae. Pulmonary sphingosine-1-phosphate levels, messenger RNA expression, and permeability as well as lung morphology were analyzed. Human blood-derived macrophages and human pulmonary microvascular endothelial cells were infected with S. pneumoniae. Transcellular electrical resistance of human pulmonary microvascular endothelial cell monolayers was examined. Further, permeability of murine isolated perfused lungs was determined following exposition to sphingosine-1-phosphate and pneumolysin. Measurements and Main Results: Following S. pneumoniae infection, murine pulmonary sphingosine-1-phosphate levels and sphingosine kinase 1 and sphingosine-1-phosphate receptor 2 expression were increased. Pneumonia-induced lung hyperpermeability was reduced in SphK1(-/-) mice compared with wild-type mice. Expression of sphingosine kinase 1 in macrophages recruited to inflamed lung areas in pneumonia was observed in murine and human lungs. S. pneumoniae induced the sphingosine kinase 1/sphingosine-1-phosphate system in blood-derived macrophages and enhanced sphingosine-1-phosphate receptor 2 expression in human pulmonary microvascular endothelial cell in vitro. In isolated mouse lungs, pneumolysin-induced hyperpermeability was dose dependently and synergistically increased by sphingosine-1-phosphate. This sphingosine-1-phosphate-induced increase was reduced by inhibition of sphingosine-1-phosphate receptor 2 or its downstream effector Rho-kinase. Conclusions: Our data suggest that targeting the sphingosine kinase 1-/sphingosine-1-phosphate-/sphingosine-1-phosphate receptor 2-signaling pathway in the lung may provide a novel therapeutic perspective in pneumococcal pneumonia for prevention of acute lung injury.
KW  - acute lung injury
KW  - pneumococcal pneumonia
KW  - sphingosine kinase 1
KW  - sphingosine-1-phosphate
KW  - sphingosine-1-phosphate receptor 2
Y1  - 2018
U6  - https://doi.org/10.1097/CCM.0000000000002916
SN  - 0090-3493
SN  - 1530-0293
VL  - 46
IS  - 3
SP  - e258
EP  - e267
PB  - Lippincott Williams & Wilkins
CY  - Philadelphia
ER  - 
TY  - JOUR
A1  - Balzus, Benjamin
A1  - Sahle, Fitsum Feleke
A1  - Hönzke, Stefan
A1  - Gerecke, Christian
A1  - Schumacher, Fabian
A1  - Hedtrich, Sarah
A1  - Kleuser, Burkhard
A1  - Bodmeier, Roland
T1  - Formulation and ex vivo evaluation of polymeric nanoparticles for controlled delivery of corticosteroids to the skin and the corneal epithelium
JF  - European journal of pharmaceutics and biopharmaceutics : EJPB ; official journal of the International Association for Pharmaceutical Technology
N2  - Controlled delivery of corticosteroids using nanoparticles to the skin and corneal epithelium may reduce their side effects and maximize treatment effectiveness. Dexamethasone-loaded ethyl cellulose, Eudragit® RS and ethyl cellulose/Eudragit® RS nanoparticles were prepared by the solvent evaporation method. Dexamethasone release from the polymeric nanoparticles was investigated in vitro using Franz diffusion cells. Drug penetration was also assessed ex vivo using excised human skin. Nanoparticle toxicity was determined by MTT and H2DCFDA assays. Eudragit® RS nanoparticles were smaller and positively charged but had a lower dexamethasone loading capacity (0.3–0.7%) than ethyl cellulose nanoparticles (1.4–2.2%). By blending the two polymers (1:1), small (105 nm), positively charged (+37 mV) nanoparticles with sufficient dexamethasone loading (1.3%) were obtained. Dexamethasone release and penetration significantly decreased with decreasing drug to polymer ratio and increased when Eudragit® RS was blended with ethyl cellulose. Ex vivo, drug release and penetration from the nanoparticles was slower than a conventional cream. The nanoparticles bear no toxicity potentials except ethyl cellulose nanoparticles had ROS generation potential at high concentration. In conclusion, the nanoparticles showed great potential to control the release and penetration of corticosteroids on the skin and mucus membrane and maximize treatment effectiveness.
KW  - Dermal delivery
KW  - Dexamethasone
KW  - Ethyl cellulose
KW  - Eudragit (R) RS
KW  - Ocular delivery
KW  - Polymeric nanoparticle
Y1  - 2017
U6  - https://doi.org/10.1016/j.ejpb.2017.02.001
SN  - 0939-6411
SN  - 1873-3441
VL  - 115
SP  - 122
EP  - 130
PB  - Elsevier
CY  - Amsterdam
ER  - 
TY  - GEN
A1  - Pischon, Hannah
A1  - Radbruch, Moritz
A1  - Ostrowski, Anja
A1  - Schumacher, Fabian
A1  - Hoenzke, Stefan
A1  - Kleuser, Burkhard
A1  - Hedtrich, Sarah
A1  - Fluhr, Joachim W.
A1  - Gruber, Achim D.
A1  - Mundhenk, Lars
T1  - How Effective Is Tacrolimus in the Imiquimod
BT  - Induced Mouse Model of Psoriasis?
T2  - The journal of investigative dermatology
Y1  - 2017
U6  - https://doi.org/10.1016/j.jid.2017.09.019
SN  - 0022-202X
SN  - 1523-1747
VL  - 138
IS  - 2
SP  - 455
EP  - 458
PB  - Elsevier
CY  - New York
ER  - 
TY  - JOUR
A1  - Wiebeler, Christian
A1  - Vollbrecht, Joachim
A1  - Neuba, Adam
A1  - Kitzerow, Heinz
A1  - Schumacher, Stefan
T1  - Unraveling the electrochemical and spectroscopic properties of neutral and negatively charged perylene tetraethylesters
JF  - Scientific reports
N2  - A detailed investigation of the energy levels of perylene-3,4,9,10-tetracarboxylic tetraethylester as a representative compound for the whole family of perylene esters was performed. It was revealed via electrochemical measurements that one oxidation and two reductions take place. The bandgaps determined via the electrochemical approach are in good agreement with the optical bandgap obtained from the absorption spectra via a Tauc plot. In addition, absorption spectra in dependence of the electrochemical potential were the basis for extensive quantum-chemical calculations of the neutral, monoanionic, and dianionic molecules. For this purpose, calculations based on density functional theory were compared with post-Hartree-Fock methods and the CAM-B3LYP functional proved to be the most reliable choice for the calculation of absorption spectra. Furthermore, spectral features found experimentally could be reproduced with vibronic calculations and allowed to understand their origins. In particular, the two lowest energy absorption bands of the anion are not caused by absorption of two distinct electronic states, which might have been expected from vertical excitation calculations, but both states exhibit a strong vibronic progression resulting in contributions to both bands.
Y1  - 2021
U6  - https://doi.org/10.1038/s41598-021-95551-0
SN  - 2045-2322
VL  - 11
IS  - 1
PB  - Macmillan Publishers Limited, part of Springer Nature
CY  - London
ER  -