227 results on '"Edgar Schmitt"'
Search Results
2. Liver-Primed Memory T Cells Generated under Noninflammatory Conditions Provide Anti-infectious Immunity
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Jan P. Böttcher, Oliver Schanz, Dirk Wohlleber, Zeinab Abdullah, Svenja Debey-Pascher, Andrea Staratschek-Jox, Bastian Höchst, Silke Hegenbarth, Jessica Grell, Andreas Limmer, Imke Atreya, Markus F. Neurath, Dirk H. Busch, Edgar Schmitt, Peter van Endert, Waldemar Kolanus, Christian Kurts, Joachim L. Schultze, Linda Diehl, and Percy A. Knolle
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Biology (General) ,QH301-705.5 - Abstract
Development of CD8+ T cell (CTL) immunity or tolerance is linked to the conditions during T cell priming. Dendritic cells (DCs) matured during inflammation generate effector/memory T cells, whereas immature DCs cause T cell deletion/anergy. We identify a third outcome of T cell priming in absence of inflammation enabled by cross-presenting liver sinusoidal endothelial cells. Such priming generated memory T cells that were spared from deletion by immature DCs. Similar to central memory T cells, liver-primed T cells differentiated into effector CTLs upon antigen re-encounter on matured DCs even after prolonged absence of antigen. Their reactivation required combinatorial signaling through the TCR, CD28, and IL-12R and controlled bacterial and viral infections. Gene expression profiling identified liver-primed T cells as a distinct Neuropilin-1+ memory population. Generation of liver-primed memory T cells may prevent pathogens that avoid DC maturation by innate immune escape from also escaping adaptive immunity through attrition of the T cell repertoire.
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- 2013
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3. Combined B, T and NK Cell Deficiency Accelerates Atherosclerosis in BALB/c Mice.
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Fei Cheng, Laura Twardowski, Kurt Reifenberg, Kerstin Winter, Antje Canisius, Eva Pross, Jianglin Fan, Edgar Schmitt, Leonard D Shultz, Karl J Lackner, and Michael Torzewski
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Medicine ,Science - Abstract
This study focused on the unique properties of both the Ldlr knockout defect (closely mimicking the human situation) and the BALB/c (C) inbred mouse strain (Th-2 slanted immune response). We generated two immunodeficient strains with severe combined B- and T-cell immunodeficiency with or without a complete lack of natural killer cells to revisit the role of adaptive immune responses on atherogenesis. C-Ldlr-/- Rag1-/- mice, which show severe combined B- and T-cell immunodeficiency and C-Ldlr-/- Rag1-/- Il2rg-/- mice, which combine the T- and B-cell defect with a complete lack of natural killer cells and inactivation of multiple cytokine signalling pathways were fed an atherogenic Western type diet (WTD). Both B6-Ldlr-/- and C-Ldlr-/- immunocompetent mice were used as controls. Body weights and serum cholesterol levels of both immunodeficient strains were significantly increased compared to C-Ldlr-/- controls, except for cholesterol levels of C-Ldlr-/- Rag1-/- double mutants after 12 weeks on the WTD. Quantification of the aortic sinus plaque area revealed that both strains of immunodeficient mice developed significantly more atherosclerosis compared to C-Ldlr-/- controls after 24 weeks on the WTD. Increased atherosclerotic lesion development in C-Ldlr-/- Rag1-/- Il2rg-/- triple mutants was associated with significantly increased numbers of macrophages and significantly decreased numbers of smooth muscle cells compared to both C-Ldlr-/- wild type and C-Ldlr-/- Rag1-/- double mutants pointing to a plaque destabilizing effect of NK cell loss. Collectively, the present study reveals a previously unappreciated complexity with regard to the impact of lymphocytes on lipoprotein metabolism and the role of lymphocyte subsets in plaque composition.
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- 2016
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4. Mechanisms of cyclic nucleotide phosphodiesterases in modulating T cell responses in murine graft-versus-host disease.
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Michael Weber, Corinna Lupp, Pamela Stein, Andreas Kreft, Tobias Bopp, Thomas C Wehler, Edgar Schmitt, Hansjörg Schild, and Markus P Radsak
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Medicine ,Science - Abstract
Graft-versus-host disease (GvHD) is a key contributor to the morbidity and mortality after allogeneic hematopoetic stem cell transplantation (HSCT). Regulatory Foxp3(+) CD4(+) T cells (Treg) suppress conventional T cell activation and can control GvHD. In our previous work, we demonstrate that a basic mechanism of Treg mediated suppression occurs by the transfer of cyclic adenosine monophosphate (cAMP) to responder cells. Whether this mechanism is relevant for Treg mediated suppression of GvHD is currently unknown. To address this question, bone marrow and T cells from C57BL/6 mice were transferred into lethally irradiated BALB/c recipients, and the course of GvHD and survival were monitored. Transplanted recipients developed severe GvHD that was strongly ameliorated by the transfer of donor Treg cells. Towards the underlying mechanisms, in vitro studies revealed that Treg communicated with DCs via gap junctions, resulting in functional inactivation of DC by a metabolic pathway involving cAMP that is modulated by the phosphodiesterase (PDE) 4 inhibitor rolipram. PDE2 or PDE3 inhibitors as well as rolipram suppressed allogeneic T cell activation, indirectly by enhancing Treg mediated suppression of DC activation and directly by inhibiting responder T cell proliferation. In line with this, we observed a cooperative suppression of GvHD upon Treg transfer and additional rolipram treatment. In conclusion, we propose that an important pathway of Treg mediated control of GvHD is based on a cAMP dependent mechanism. These data provide the basis for future concepts to manipulate allogeneic T cell responses to prevent GvHD.
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- 2013
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5. DR(high+)CD45RA(-)-Tregs potentially affect the suppressive activity of the total Treg pool in renal transplant patients.
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Matthias Schaier, Nicole Seissler, Edgar Schmitt, Stefan Meuer, Friederike Hug, Martin Zeier, and Andrea Steinborn
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Medicine ,Science - Abstract
Recent studies show that regulatory T cells (Tregs) play an essential role in tolerance induction after organ transplantation. In order to examine whether there are differences in the composition of the total CD4(+)CD127(low+/-)FoxP3(+)- Treg cell pool between stable transplant patients and patients with biopsy proven rejection (BPR), we compared the percentages and the functional activity of the different Treg cell subsets (DR(high+)CD45RA(-)-Tregs, DR(low+)CD45RA(-)-Tregs, DR(-)CD45RA(-)-Tregs, DR(-)CD45RA(+)-Tregs). All parameters were determined during the three different periods of time after transplantation (0-30 days, 31-1,000 days, >1,000 days). Among 156 transplant patients, 37 patients suffered from BPR. The most prominent differences between rejecting and non-rejecting patients were observed regarding the DR(high+)CD45RA(-)-Treg cell subset. Our data demonstrate that the suppressive activity of the total Treg pool strongly depends on the presence of these Treg cells. Their percentage within the total Treg pool strongly decreased after transplantation and remained relatively low during the first year after transplantation in all patients. Subsequently, the proportion of this Treg subset increased again in patients who accepted the transplant and reached a value of healthy non-transplanted subjects. By contrast, in patients with acute kidney rejection, the DR(high+)CD45RA(-)-Treg subset disappeared excessively, causing a reduction in the suppressive activity of the total Treg pool. Therefore, both the monitoring of its percentage within the total Treg pool and the monitoring of the HLA-DR MFI of the DR(+)CD45RA(-)-Treg subset may be useful tools for the prediction of graft rejection.
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- 2012
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6. Regulatory T cells and IL-10 independently counterregulate cytotoxic T lymphocyte responses induced by transcutaneous immunization.
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Pamela Stein, Michael Weber, Steve Prüfer, Beate Schmid, Edgar Schmitt, Hans-Christian Probst, Ari Waisman, Peter Langguth, Hansjörg Schild, and Markus P Radsak
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Medicine ,Science - Abstract
The imidazoquinoline derivate imiquimod induces inflammatory responses and protection against transplanted tumors when applied to the skin in combination with a cognate peptide epitope (transcutaneous immunization, TCI). Here we investigated the role of regulatory T cells (T(reg)) and the suppressive cytokine IL-10 in restricting TCI-induced cytotoxic T lymphocyte (CTL) responses.TCI was performed with an ointment containing the TLR7 agonist imiquimod and a CTL epitope was applied to the depilated back skin of C57BL/6 mice. Using specific antibodies and FoxP3-diphteria toxin receptor transgenic (DEREG) mice, we interrogated inhibiting factors after TCI: by depleting FoxP3(+) regulatory T cells we found that specific CTL-responses were greatly enhanced. Beyond this, in IL-10 deficient (IL-10(-/-)) mice or after blocking of IL-10 signalling with an IL-10 receptor specific antibody, the TCI induced CTL response is greatly enhanced indicating an important role for this cytokine in TCI. However, by transfer of T(reg) in IL-10(-/-) mice and the use of B cell deficient JHT(-/-) mice, we can exclude T(reg) and B cells as source of IL-10 in the setting of TCI.We identify T(reg) and IL-10 as two important and independently acting suppressors of CTL-responses induced by transcutaneous immunization. Advanced vaccination strategies inhibiting T(reg) function and IL-10 release may lead the development of effective vaccination protocols aiming at the induction of T cell responses suitable for the prophylaxis or treatment of persistent infections or tumors.
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- 2011
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7. miR-155 inhibition sensitizes CD4+ Th cells for TREG mediated suppression.
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Heiko F Stahl, Tanja Fauti, Nina Ullrich, Tobias Bopp, Jan Kubach, Werner Rust, Paul Labhart, Vassili Alexiadis, Christian Becker, Mathias Hafner, Andreas Weith, Martin C Lenter, Helmut Jonuleit, Edgar Schmitt, and Detlev Mennerich
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Medicine ,Science - Abstract
BackgroundIn humans and mice naturally occurring CD4(+)CD25(+) regulatory T cells (nTregs) are a thymus-derived subset of T cells, crucial for the maintenance of peripheral tolerance by controlling not only potentially autoreactive T cells but virtually all cells of the adaptive and innate immune system. Recent work using Dicer-deficient mice irrevocably demonstrated the importance of miRNAs for nTreg cell-mediated tolerance.Principal findingsDNA-Microarray analyses of human as well as murine conventional CD4(+) Th cells and nTregs revealed a strong up-regulation of mature miR-155 (microRNA-155) upon activation in both populations. Studying miR-155 expression in FoxP3-deficient scurfy mice and performing FoxP3 ChIP-Seq experiments using activated human T lymphocytes, we show that the expression and maturation of miR-155 seem to be not necessarily regulated by FoxP3. In order to address the functional relevance of elevated miR-155 levels, we transfected miR-155 inhibitors or mature miR-155 RNAs into freshly-isolated human and mouse primary CD4(+) Th cells and nTregs and investigated the resulting phenotype in nTreg suppression assays. Whereas miR-155 inhibition in conventional CD4(+) Th cells strengthened nTreg cell-mediated suppression, overexpression of mature miR-155 rendered these cells unresponsive to nTreg cell-mediated suppression.ConclusionInvestigation of FoxP3 downstream targets, certainly of bound and regulated miRNAs revealed the associated function between the master regulator FoxP3 and miRNAs as regulators itself. miR-155 is shown to be crucially involved in nTreg cell mediated tolerance by regulating the susceptibility of conventional human as well as murine CD4(+) Th cells to nTreg cell-mediated suppression.
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- 2009
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8. Epigenetic control of the foxp3 locus in regulatory T cells.
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Stefan Floess, Jennifer Freyer, Christiane Siewert, Udo Baron, Sven Olek, Julia Polansky, Kerstin Schlawe, Hyun-Dong Chang, Tobias Bopp, Edgar Schmitt, Stefan Klein-Hessling, Edgar Serfling, Alf Hamann, and Jochen Huehn
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Biology (General) ,QH301-705.5 - Abstract
Compelling evidence suggests that the transcription factor Foxp3 acts as a master switch governing the development and function of CD4(+) regulatory T cells (Tregs). However, whether transcriptional control of Foxp3 expression itself contributes to the development of a stable Treg lineage has thus far not been investigated. We here identified an evolutionarily conserved region within the foxp3 locus upstream of exon-1 possessing transcriptional activity. Bisulphite sequencing and chromatin immunoprecipitation revealed complete demethylation of CpG motifs as well as histone modifications within the conserved region in ex vivo isolated Foxp3(+)CD25(+)CD4(+) Tregs, but not in naïve CD25(-)CD4(+) T cells. Partial DNA demethylation is already found within developing Foxp3(+) thymocytes; however, Tregs induced by TGF-beta in vitro display only incomplete demethylation despite high Foxp3 expression. In contrast to natural Tregs, these TGF-beta-induced Foxp3(+) Tregs lose both Foxp3 expression and suppressive activity upon restimulation in the absence of TGF-beta. Our data suggest that expression of Foxp3 must be stabilized by epigenetic modification to allow the development of a permanent suppressor cell lineage, a finding of significant importance for therapeutic applications involving induction or transfer of Tregs and for the understanding of long-term cell lineage decisions.
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- 2007
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9. A flexible, modular, open-source implementation of 6LoWPAN.
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Manuel Schappacher, Edgar Schmitt, Axel Sikora, Patrick Weber, and Artem Yushev
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- 2015
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10. The Development of Vaccines from Synthetic Tumor‐Associated Mucin Glycopeptides and their Glycosylation‐Dependent Immune Response
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Pol Besenius, Moritz Urschbach, Edgar Schmitt, Natascha Stergiou, Horst Kunz, and Adele Gabba
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Glycosylation ,General Chemical Engineering ,medicine.medical_treatment ,Biology ,Cancer Vaccines ,Biochemistry ,Epitope ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Immune system ,Antigen ,Neoplasms ,Materials Chemistry ,medicine ,Humans ,MUC1 ,030304 developmental biology ,Vaccines, Synthetic ,0303 health sciences ,Mucin ,Glycopeptides ,Immunity ,Mucins ,General Chemistry ,Immunotherapy ,3. Good health ,chemistry ,030220 oncology & carcinogenesis ,Immunology ,Adjuvant - Abstract
Tumor-associated carbohydrate antigens are overexpressed as altered-self in most common epithelial cancers. Their glycosylation patterns differ from those of healthy cells, functioning as an ID for cancer cells. Scientists have been developing anti-cancer vaccines based on mucin glycopeptides, yet the interplay of delivery system, adjuvant and tumor associated MUC epitopes in the induced immune response is not well understood. The current state of the art suggests that the identity, abundancy and location of the glycans on the MUC backbone are all key parameters in the cellular and humoral response. This review shares lessons learned by us in over two decades of research in glycopeptide vaccines. By bridging synthetic chemistry and immunology, we discuss efforts in designing synthetic MUC1/4/16 vaccines and focus on the role of glycosylation patterns. We provide a brief introduction into the mechanisms of the immune system and aim to promote the development of cancer subunit vaccines.
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- 2021
11. In Activated Murine Mast Cells, NFATc2 Is Critical for the Production of Autocrine IL-3, Thereby Promoting the Expression of IL-9
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Lorenz Ullner, Matthias Klein, Jennifer Hahlbrock, Toszka Bohn, Farhad Sabbaghi, Tobias Bopp, Edgar Schmitt, and Michael Stassen
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Cell type ,NFATC2 ,medicine.medical_treatment ,Immunology ,Cell ,Autocrine Communication ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Downregulation and upregulation ,STAT5 Transcription Factor ,medicine ,Animals ,Immunology and Allergy ,Mast Cells ,Autocrine signalling ,Cells, Cultured ,STAT5 ,Feedback, Physiological ,Mice, Knockout ,Mice, Inbred BALB C ,NFATC Transcription Factors ,biology ,Chemistry ,Interleukin-9 ,T-Lymphocytes, Helper-Inducer ,Up-Regulation ,Cell biology ,Mice, Inbred C57BL ,Cytokine ,medicine.anatomical_structure ,biology.protein ,Interleukin-3 ,030215 immunology - Abstract
IL-9 has lent its numerical designation to the Th9 subset of CD4+ Th cells, although it is also produced by additional cell types, including mast cells. It is a pleiotropic cytokine involved in allergic reactions, parasitic infections, autoimmune inflammation, and cancer immunity. In this article, we provide evidence that NFATc2 has contradictory functions in the expression of IL-9 in murine Th9 cells and bone marrow–derived mast cells (BMMC). The basis for this is our observation that the production of IL-9 in NFATc2-deficient Th9 cells is increased, whereas it is decreased in BMMC devoid of NFATc2. In addition, NFATc2 deficiency almost completely abrogates the expression of IL-3 in both cell types. However, selectively in BMMC, the production of IL-9 critically depends on autocrine IL-3 acting via the sustained activation of STAT5 on the expression of IL-9. Furthermore, we demonstrate that IL-3 acts independently and synergistically with IL-1β on the production of IL-9. Taken together, we highlight NFATc2-driven production of autocrine IL-3 as a critical and cell type–specific component for IL-9 expression in BMMC.
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- 2021
12. The structure and function of Iristatin, a novel immunosuppressive tick salivary cystatin
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Edgar Schmitt, Natascha Stergiou, Helena Langhansová, Eric Calvo, Michail Kotsyfakis, Alexandra Schwarz, Jan Kotál, Michael Mareš, Zuzana Beránková, Jan Kopecký, Pavlína Řezáčová, Adéla Chlastáková, Jindřich Chmelař, and Michal Buša
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Saliva ,T-Lymphocytes ,Biology ,Crystallography, X-Ray ,Nitric Oxide ,Arthropod Proteins ,law.invention ,Cellular and Molecular Neuroscience ,Immune system ,In vivo ,law ,Animals ,Amino Acid Sequence ,Molecular Biology ,Phylogeny ,Pharmacology ,Cathepsin ,Ixodes ,Sequence Homology, Amino Acid ,Macrophages ,Cell Biology ,Cystatins ,Cysteine protease ,In vitro ,Cell biology ,Proteolysis ,Recombinant DNA ,Cytokines ,Epoxy Compounds ,Salivary Cystatins ,Tyrosine ,Molecular Medicine ,Female ,Cystatin ,Immunosuppressive Agents - Abstract
To successfully feed, ticks inject pharmacoactive molecules into the vertebrate host including cystatin cysteine protease inhibitors. However, the molecular and cellular events modulated by tick saliva remain largely unknown. Here, we describe and characterize a novel immunomodulatory cystatin, Iristatin, which is upregulated in the salivary glands of feeding Ixodes ricinus ticks. We present the crystal structure of Iristatin at 1.76 A resolution. Purified recombinant Iristatin inhibited the proteolytic activity of cathepsins L and C and diminished IL-2, IL-4, IL-9, and IFN-γ production by different T-cell populations, IL-6 and IL-9 production by mast cells, and nitric oxide production by macrophages. Furthermore, Iristatin inhibited OVA antigen-induced CD4+ T-cell proliferation and leukocyte recruitment in vivo and in vitro. Our results indicate that Iristatin affects wide range of anti-tick immune responses in the vertebrate host and may be exploitable as an immunotherapeutic.
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- 2019
13. Evaluation of a novel monoclonal antibody against tumor-associated MUC1 for diagnosis and prognosis of breast cancer
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Natascha Stergiou, Horst Kunz, Frank Roesch, Walburgis Brenner, Matthias Miederer, Edgar Schmitt, Jörg Jäkel, Stefanie Pektor, Johannes Nagel, Anne-Sophie Heimes, and Marcus Schmidt
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medicine.drug_class ,Estrogen receptor ,MUC1 ,Breast Neoplasms ,Mice, Transgenic ,Deferoxamine ,Monoclonal antibody ,89Zr ,03 medical and health sciences ,0302 clinical medicine ,Breast cancer ,In vivo ,Cell Line, Tumor ,Biomarkers, Tumor ,medicine ,Animals ,Humans ,Tissue Distribution ,skin and connective tissue diseases ,Triple-negative breast cancer ,mAb ,Radioisotopes ,Mice, Inbred BALB C ,biology ,business.industry ,Mucin-1 ,breast cancer diagnosis ,Antibodies, Monoclonal ,Cancer ,General Medicine ,Prognosis ,medicine.disease ,Immunohistochemistry ,Mice, Inbred C57BL ,Positron-Emission Tomography ,biology.protein ,Cancer research ,Female ,030211 gastroenterology & hepatology ,Zirconium ,Antibody ,business ,Research Paper - Abstract
There is still a great unmet medical need concerning diagnosis and treatment of breast cancer which could be addressed by utilizing specific molecular targets. Tumor-associated MUC1 is expressed on over 90 % of all breast cancer entities and differs strongly from its physiological form on epithelial cells, therefore presenting a unique target for breast cancer diagnosis and antibody-mediated immune therapy. Utilizing an anti-tumor vaccine based on a synthetically prepared glycopeptide, we generated a monoclonal antibody (mAb) GGSK-1/30, selectively recognizing human tumor-associated MUC1. This antibody targets exclusively tumor-associated MUC1 in the absence of any binding to MUC1 on healthy epithelial cells thus enabling the generation of breast tumor-specific radiolabeled immune therapeutic tools. Methods: MAb GGSK-1/30 was used for immunohistochemical analysis of human breast cancer tissue. Its desferrioxamine (Df')-conjugate was synthesized and labelled with 89Zr. [89Zr]Zr-Df'-GGSK-1/30 was evaluated as a potential PET tracer. Binding and pharmacokinetic properties of [89Zr]Zr-Df'-GGSK-1/30 were analyzed in vitro using human and murine cell lines that express tumor-associated MUC1. Self-generated primary murine breast cancer cells expressing human tumor-associated MUC1 were transplanted subcutaneously in wild type and human MUC1-transgenic mice. The pharmacology of [89Zr]Zr-Df'-GGSK-1/30 was investigated using breast tumor-bearing mice in vivo by PET/MRT imaging as well as by ex vivo organ biodistribution analysis. Results: The mAb GGSK-1/30 stained specifically human breast tumor tissue and can be possibly used to predict the severity of disease progression based on the expression of the tumor-associated MUC1. For in vivo imaging, the Df'-conjugated mAb was radiolabeled with a radiochemical yield of 60 %, a radiochemical purity of 95 % and an apparent specific activity of 6.1 GBq/µmol. After 7 d, stabilities of 84 % in human serum and of 93 % in saline were observed. In vitro cell studies showed strong binding to human tumor-associated MUC1 expressing breast cancer cells. The breast tumor-bearing mice showed an in vivo tumor uptake of >50 %ID/g and clearly visible specific enrichment of the radioconjugate via PET/MRT. Principal conclusions: Tumor-associated MUC1 is a very important biomarker for breast cancer next to the traditional markers estrogen receptor (ER), progesterone receptor (PR) and HER/2-neu. The mAb GGSK-1/30 can be used for the diagnosis of over 90% of breast cancers, including triple negative breast cancer based on biopsy staining. Its radioimmunoconjugate represents a promising PET-tracer for breast cancer imaging selectively targeting breast cancer cells.
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- 2019
14. (TA)MUC1 as a potential new target for breast cancer therapy
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Annette Hasenburg, Walburgis Brenner, Anne-Sophie Heimes, Edgar Schmitt, N Stergiou, R Attariya, P Fries, and Martina Schmidt
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Oncology ,medicine.medical_specialty ,Breast cancer ,business.industry ,Internal medicine ,Medicine ,business ,medicine.disease ,MUC1 - Published
- 2020
15. Synthetic MUC1 Antitumor Vaccine with Incorporated 2,3-Sialyl-T Carbohydrate Antigen Inducing Strong Immune Responses with Isotype Specificity
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Sabrina Bialas, Natascha Stergiou, Horst Kunz, David Straßburger, Pol Besenius, Markus Glaffig, and Edgar Schmitt
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0301 basic medicine ,Glycosylation ,Chemistry ,medicine.medical_treatment ,Organic Chemistry ,Toxoid ,010402 general chemistry ,01 natural sciences ,Biochemistry ,Isotype ,Molecular biology ,0104 chemical sciences ,03 medical and health sciences ,chemistry.chemical_compound ,030104 developmental biology ,Immune system ,Antigen ,Peptide vaccine ,medicine ,Molecular Medicine ,Molecular Biology ,Adjuvant ,MUC1 - Abstract
The endothelial glycoprotein MUC1 is known to underlie alterations in cancer by means of aberrant glycosylation accompanied by changes in morphology. The heavily shortened glycans induce a collapse of the peptide backbone and enable accessibility of the latter to immune cells, rendering it a tumor-associated antigen. Synthetic vaccines based on MUC1 tandem repeat motifs, comprising tumor-associated 2,3-sialyl-T antigen, conjugated to the immunostimulating tetanus toxoid, are reported herein. Immunization with these vaccines in a simple water/oil emulsion produced a strong immune response in mice to which stimulation with complete Freund's adjuvant (CFA) was not superior. In both cases, high levels of IgG1 and IgG2a/b were induced in C57BL/6 mice. Additional glycosylation in the immunodominant PDTRP domain led to improved binding of the induced antisera to MCF-7 breast tumor cells, compared with that of the monoglycosylated peptide vaccine.
- Published
- 2018
16. Mannose-Decorated Multicomponent Supramolecular Polymers Trigger Effective Uptake into Antigen-Presenting Cells
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Hajime Yurugi, Edgar Schmitt, Daniel Spitzer, Natascha Stergiou, David Straßburger, Dieter Schollmeyer, Pol Besenius, and Moritz Urschbach
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Models, Molecular ,Dendrimers ,Mannosides ,Biocompatibility ,Supramolecular chemistry ,Antigen-Presenting Cells ,010402 general chemistry ,01 natural sciences ,Biochemistry ,Polyethylene Glycols ,Mice ,Surface-Active Agents ,chemistry.chemical_compound ,Amphiphile ,Animals ,Antigen-presenting cell ,Molecular Biology ,Cells, Cultured ,Fluorescent Dyes ,chemistry.chemical_classification ,Microscopy, Confocal ,010405 organic chemistry ,Macrophages ,Organic Chemistry ,Biological Transport ,Carbocyanines ,Cell sorting ,0104 chemical sciences ,Supramolecular polymers ,chemistry ,Biophysics ,Molecular Medicine ,Peptides ,Ethylene glycol - Abstract
A modular route to prepare functional self-assembling dendritic peptide amphiphiles decorated with mannosides, to effectively target antigen-presenting cells, such as macrophages, is reported. The monomeric building blocks were equipped with tetra(ethylene glycol)s (TEGs) or labeled with a Cy3 fluorescent probe. Experiments on the uptake of the multifunctional supramolecular particles into murine macrophages (Mφs) were monitored by confocal microscopy and fluorescence-activated cell sorting. Mannose-decorated supramolecular polymers trigger a significantly higher cellular uptake and distribution, relative to TEG carrying bare polymers. No cytotoxicity or negative impact on cytokine production of the treated Mφs was observed, which emphasized their biocompatibility. The modular nature of the multicomponent supramolecular polymer coassembly protocol is a promising platform to develop fully synthetic multifunctional vaccines, for example, in cancer immunotherapy.
- Published
- 2018
17. A Synthetic MUC1 Anticancer Vaccine Containing Mannose Ligands for Targeting Macrophages and Dendritic Cells
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Edgar Schmitt, Natascha Stergiou, Sebastian Hartmann, Horst Kunz, and Markus Glaffig
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CD4-Positive T-Lymphocytes ,0301 basic medicine ,medicine.medical_treatment ,Mannose ,Enzyme-Linked Immunosorbent Assay ,Receptors, Cell Surface ,Ligands ,010402 general chemistry ,Cancer Vaccines ,01 natural sciences ,Biochemistry ,Divalent ,Mice ,03 medical and health sciences ,chemistry.chemical_compound ,Immune system ,Cancer immunotherapy ,Drug Discovery ,medicine ,Animals ,Humans ,Lectins, C-Type ,General Pharmacology, Toxicology and Pharmaceutics ,MUC1 ,Pharmacology ,chemistry.chemical_classification ,Mice, Inbred BALB C ,biology ,Chemistry ,Macrophages ,Mucin-1 ,Organic Chemistry ,Dendritic Cells ,Molecular biology ,0104 chemical sciences ,Mannose-Binding Lectins ,030104 developmental biology ,Lymphatic system ,Immunoglobulin G ,Immunology ,MCF-7 Cells ,biology.protein ,Molecular Medicine ,Lymph Nodes ,Antibody ,Mannose Receptor ,Mannose receptor ,Protein Binding - Abstract
A MUC1 anticancer vaccine equipped with covalently linked divalent mannose ligands was found to improve the antigen uptake and presentation by targeting mannose-receptor-positive macrophages and dendritic cells. It induced much stronger specific IgG immune responses in mice than the non-mannosylated reference vaccine. Mannose coupling also led to increased numbers of macrophages, dendritic cells, and CD4+ T cells in the local lymph organs. Comparison of di- and tetravalent mannose ligands revealed an increased binding of the tetravalent version, suggesting that higher valency improves binding to the mannose receptor. The mannose-coupled vaccine and the non-mannosylated reference vaccine induced IgG antibodies that exhibited similar binding to human breast tumor cells.
- Published
- 2017
18. Liver sinusoidal endothelial cell cross-priming is supported by CD4 T cell-derived IL-2
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Christoph Garbers, Christian Kurts, Edgar Schmitt, Oliver Schanz, Silke Hegenbarth, Dirk Wohlleber, Michael Dudek, Tobias Bopp, Stefan Rose John, Percy A. Knolle, Jan P. Böttcher, and Michaela Wittlich
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CD4-Positive T-Lymphocytes ,0301 basic medicine ,T cell ,Antigen-Presenting Cells ,Priming (immunology) ,Cell Communication ,CD8-Positive T-Lymphocytes ,Biology ,Granzymes ,GZMB ,Mice ,03 medical and health sciences ,Interleukin 21 ,Cross-Priming ,0302 clinical medicine ,medicine ,Animals ,Cytotoxic T cell ,Antigen-presenting cell ,Cells, Cultured ,Hepatology ,Endothelial Cells ,T helper cell ,Natural killer T cell ,Cell biology ,Mice, Inbred C57BL ,030104 developmental biology ,medicine.anatomical_structure ,Liver ,030220 oncology & carcinogenesis ,Interleukin-2 - Abstract
Background & Aims Liver sinusoidal endothelial cells (LSECs) are prominent liver-resident antigen (cross-)presenting cells. LSEC cross-priming of naive CD8 T cells does not require CD4 T cell help in contrast to priming by dendritic cells (DC) but leads to the formation of memory T cells that is preceded by transient Granzyme B (GzmB) expression. Here we provide evidence for a so far unrecognized CD4 T helper cell function in LSEC-induced CD8 T cell activation. Methods Naive CD8 T cells and differentiated T helper 1 (T h 1) cells were stimulated by antigen-presenting LSEC, and GzmB expression in CD8 T cells was determined by flow cytometry. To identify molecular pathways mediating this GzmB expression, mechanistic proof-of-concept experiments were conducted using stimulatory anti-CD3 antibody together with Hyper-IL-6. Results We demonstrate that LSECs simultaneously function in antigen co-presentation to CD8 and CD4 T cells. Such co-presentation revealed a function of T h 1 cells to increase GzmB expression in CD8 T cells after LSEC but not DC cross-priming. IL-2 released from T h 1 cells was required but not sufficient for rapid GzmB induction in CD8 T cells. T cell receptor together with IL-6 trans-signaling was necessary for IL-2 to mediate rapid GzmB induction. Conclusions Our findings indicate that LSECs can serve as a platform to facilitate CD4–CD8 T cell crosstalk enhancing the immune function of LSECs to cross-prime CD8 T cells. IL-6 trans-signaling-mediated responsiveness for IL-2 inducing sustained GzmB expression in CD8 T cells reveals unique mechanisms of CD4 T cell help and CD8 T cell differentiation through liver-resident antigen-presenting cells. Lay summary Our findings demonstrate that LSEC co-present antigen to CD8 and CD4 T cells and thereby enable CD4 T cell help for LSEC-priming of CD8 T cells. This CD4 T cell help selectively enhances the rapid upregulation of GzmB and effector function of LSEC-primed CD8 T cells thereby enhancing functional differentiation towards CD8 effector T cells.
- Published
- 2017
19. Protein kinase CK2 governs the molecular decision between encephalitogenic T H 17 cell and T reg cell development
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Felix Luessi, Natascha Stergiou, Till-Julius Brühl, Toszka Bohn, Frauke Zipp, Tobias Bopp, Ari Waisman, Ulrike Bühler, Hansjörg Schild, Matthias Klein, Beatrice Wasser, Alexander Ulges, Edgar Schmitt, Georg Bündgen, Esther Witsch, Katharina Birkner, Gautam Pramanik, Sarah Dietzen, and Horst Kunz
- Subjects
STAT3 Transcription Factor ,0301 basic medicine ,Encephalomyelitis, Autoimmune, Experimental ,Multiple Sclerosis ,Cell ,Mice, Transgenic ,chemical and pharmacologic phenomena ,Biology ,Severity of Illness Index ,T-Lymphocytes, Regulatory ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Immune system ,medicine ,Animals ,Humans ,IL-2 receptor ,Phosphorylation ,Casein Kinase II ,STAT3 ,Multidisciplinary ,Cell growth ,Interleukin-17 ,Experimental autoimmune encephalomyelitis ,Granulocyte-Macrophage Colony-Stimulating Factor ,FOXP3 ,Peripheral tolerance ,Forkhead Transcription Factors ,hemic and immune systems ,Receptors, Interleukin ,medicine.disease ,Peptide Fragments ,Mice, Inbred C57BL ,030104 developmental biology ,medicine.anatomical_structure ,Gene Expression Regulation ,Immunology ,biology.protein ,Cancer research ,Th17 Cells ,Myelin-Oligodendrocyte Glycoprotein ,Signal Transduction ,030215 immunology - Abstract
T helper 17 (TH17) cells represent a discrete TH cell subset instrumental in the immune response to extracellular bacteria and fungi. However, TH17 cells are considered to be detrimentally involved in autoimmune diseases like multiple sclerosis (MS). In contrast to TH17 cells, regulatory T (Treg) cells were shown to be pivotal in the maintenance of peripheral tolerance. Thus, the balance between Treg cells and TH17 cells determines the severity of a TH17 cell-driven disease and therefore is a promising target for treating autoimmune diseases. However, the molecular mechanisms controlling this balance are still unclear. Here, we report that pharmacological inhibition as well as genetic ablation of the protein kinase CK2 (CK2) ameliorates experimental autoimmune encephalomyelitis (EAE) severity and relapse incidence. Furthermore, CK2 inhibition or genetic ablation prevents TH17 cell development and promotes the generation of Treg cells. Molecularly, inhibition of CK2 leads to reduced STAT3 phosphorylation and strongly attenuated expression of the IL-23 receptor, IL-17, and GM-CSF. Thus, these results identify CK2 as a nodal point in TH17 cell development and suggest this kinase as a potential therapeutic target to treat TH17 cell-driven autoimmune responses.
- Published
- 2016
20. Glycopeptide-functionalized gold nanoparticles for antibody induction against the tumor associated mucin-1 glycoprotein
- Author
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Ulrika Westerlind, Horst Kunz, Roberto Fiammengo, Bastian Gerlitzki, Federica Degliangeli, Hui Cai, Edgar Schmitt, and Björn Palitzsch
- Subjects
Genes, MHC Class II ,Molecular Sequence Data ,Clinical Biochemistry ,Epitopes, T-Lymphocyte ,Metal Nanoparticles ,Pharmaceutical Science ,02 engineering and technology ,010402 general chemistry ,Cancer Vaccines ,01 natural sciences ,Biochemistry ,Antibodies ,Epitope ,Mice ,Immune system ,Neoplasms ,Drug Discovery ,Animals ,Humans ,Amino Acid Sequence ,skin and connective tissue diseases ,Molecular Biology ,MUC1 ,Cancer ,chemistry.chemical_classification ,Antiserum ,Vaccines ,biology ,Mucin-1 ,Organic Chemistry ,Glycopeptides ,021001 nanoscience & nanotechnology ,Molecular biology ,Glycopeptide ,0104 chemical sciences ,chemistry ,Colloidal gold ,Immunology ,MCF-7 Cells ,biology.protein ,Nanoparticles ,Molecular Medicine ,Immunization ,Gold ,Antibody ,0210 nano-technology ,Glycoprotein - Abstract
We report the preparation of gold nanoparticle (AuNP)-based vaccine candidates against the tumor-associated form of the mucin-1 (MUC1) glycoprotein. Chimeric peptides, consisting of a glycopeptide sequence derived from MUC1 and the T-cell epitope P30 sequence were immobilized on PEGylated AuNPs and the ability to induce selective antibodies in vivo was investigated. After immunization, mice showed significant MHC-II mediated immune responses and their antisera recognized human MCF-7 breast cancer cells. Nanoparticles designed according to this report may become key players in the development of anticancer vaccines.
- Published
- 2016
21. Ein durch eine synthetische Glycopeptid-Vakzine induzierter monoklonaler Antiköper unterscheidet normale von malignen Brustzellen und ermöglicht die Diagnose von humanen Pankreaskarzinomen
- Author
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Edgar Schmitt, Horst Kunz, Peer Flemming, Bastian Gerlitzki, Sebastian Hartmann, Nikola Gaidzik, Nicole Teusch, Sonja Stahn, Björn Palitzsch, and Natascha Stergiou
- Subjects
010405 organic chemistry ,Chemistry ,General Medicine ,010402 general chemistry ,01 natural sciences ,0104 chemical sciences - Published
- 2016
22. Tumor immunoevasion via acidosis-dependent induction of regulatory tumor-associated macrophages
- Author
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Dennis Vogel, Sabine Muth, Jochem Koenig, Danielle Arnold-Schild, Vanessa Popp, Jacques Pouysségur, Natascha Luther, Jennifer Hahlbrock, Steffen Rapp, Hans Christian Probst, Magdalena Huber, Pamela Aranda Lopez, Marina Kreutz, Kathrin Renner, Esther von Stebut, Toszka Bohn, Christina Lueckel, Shogo Endo, Stefanie Pektor, Tobias Bopp, Till-Julius Bruehl, Matthias Klein, Christian Becker, Hansjoerg Schild, Katharina Gerlach, Nobuhiko Kojima, Almut Brand, Benno Weigmann, and Edgar Schmitt
- Subjects
0301 basic medicine ,Immunology ,Mice, Transgenic ,Biology ,Adenocarcinoma ,03 medical and health sciences ,Mice ,0302 clinical medicine ,Immune system ,medicine ,Tumor Microenvironment ,Immunology and Allergy ,Animals ,Humans ,Tumor growth ,Glycolysis ,Melanoma ,Acidosis ,Tumor microenvironment ,Macrophages ,Phenotype ,Mice, Inbred C57BL ,030104 developmental biology ,030220 oncology & carcinogenesis ,Colonic Neoplasms ,Molecular mechanism ,Transcriptional Repressor ,Cancer research ,Tumor Escape ,medicine.symptom - Abstract
Many tumors evolve sophisticated strategies to evade the immune system, and these represent major obstacles for efficient antitumor immune responses. Here we explored a molecular mechanism of metabolic communication deployed by highly glycolytic tumors for immunoevasion. In contrast to colon adenocarcinomas, melanomas showed comparatively high glycolytic activity, which resulted in high acidification of the tumor microenvironment. This tumor acidosis induced Gprotein-coupled receptor-dependent expression of the transcriptional repressor ICER in tumor-associated macrophages that led to their functional polarization toward a non-inflammatory phenotype and promoted tumor growth. Collectively, our findings identify a molecular mechanism of metabolic communication between non-lymphoid tissue and the immune system that was exploited by high-glycolytic-rate tumors for evasion of the immune system.
- Published
- 2017
23. Antibody Induction Directed against the Tumor-Associated MUC4 Glycoprotein
- Author
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Edgar Schmitt, Sebastian Hartmann, Ulrika Westerlind, Björn Palitzsch, Hui Cai, Natascha Stergiou, and Horst Kunz
- Subjects
Antibodies, Neoplasm ,Molecular Sequence Data ,Cancer Vaccines ,Biochemistry ,Epitope ,Epitopes ,Mice ,Antigen ,Antibody Specificity ,Cell Line, Tumor ,Tetanus Toxoid ,Animals ,Humans ,Amino Acid Sequence ,Molecular Biology ,MUC1 ,chemistry.chemical_classification ,Mice, Inbred BALB C ,Mucin-4 ,biology ,Immune Sera ,Immunogenicity ,Vaccination ,Organic Chemistry ,Toxoid ,Glycopeptide ,Pancreatic Neoplasms ,chemistry ,Tandem Repeat Sequences ,Immunology ,biology.protein ,Molecular Medicine ,Female ,sense organs ,Antibody ,Glycoprotein - Abstract
Mucin glycoproteins are important diagnostic and therapeutic targets for cancer treatment. Although several strategies have been developed to explore anti-tumor vaccines based on MUC1 glycopeptides, only few studies have focused on vaccines directed against the tumor-associated MUC4 glycoprotein. MUC4 is an important tumor marker overexpressed in lung cancer and uniquely expressed in pancreatic ductual adenocarcinoma. The aberrant glycosylation of MUC4 in tumor cells results in an exposure of its peptide backbone and the formation of tumor-associated glycopeptide antigens. Due to the low immunogenicity of these endogenous structures, their conjugation with immune stimulating peptide or protein carriers are required. In this study, MUC4 tandem-repeat glycopeptides were conjugated to the tetanus toxoid and used for vaccination of mice. Immunological evaluations showed that our MUC4-based vaccines induced very strong antigen-specific immune responses. In addition, antibody binding epitope analysis on glycopeptide microarrays, were demonstrating a clear glycosylation site dependence of the induced antibodies.
- Published
- 2015
24. Tick saliva increases production of three chemokines including monocyte chemoattractant protein-1, a histamine-releasing cytokine
- Author
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Jan Kopecký, Helena Langhansová, Edgar Schmitt, and Tobias Bopp
- Subjects
Chemokine ,Saliva ,Ixodes ricinus ,medicine.medical_treatment ,Chemokine CXCL2 ,Immunology ,Biology ,Histamine Release ,Chemokine CCL1 ,Mice ,chemistry.chemical_compound ,Th2 Cells ,Immune system ,parasitic diseases ,medicine ,Animals ,Chemokine CCL2 ,Ixodes ,Monocyte ,Chemotaxis ,biology.organism_classification ,Specific Pathogen-Free Organisms ,Mice, Inbred C57BL ,Cytokine ,medicine.anatomical_structure ,chemistry ,biology.protein ,Female ,Parasitology ,Histamine - Abstract
Summary The effect of Ixodes ricinus tick saliva on the production of various cytokines and chemokines by mouse splenocytes was tested by a cytokine array. We demonstrated a strong upregulation of three chemokines, monocyte chemoattractant protein-1 (MCP-1), thymus-derived chemotactic agent 3 (TCA-3) and macrophage inflammatory protein 2 (MIP-2). MCP-1 could be induced by tick saliva itself. While TCA-3 and MIP-2 are engaged in Th2 polarization of the host immune response associated with tick feeding, MCP-1 may act as a histamine release factor, increasing blood flow into the feeding lesion thus facilitating tick engorgement in the late, rapid feeding phase.
- Published
- 2015
25. Protein kinase CK2 enables regulatory T cells to suppress excessive TH2 responses in vivo
- Author
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Susanne Hahn, Toszka Bohn, Hansjörg Schild, Alexander Ulges, Frauke Zipp, Nadine Grebe, Tobias Bopp, Ari Waisman, Sonja Reißig, Andreas Beilhack, Sabine Muth, Natascha Stergiou, Sebastian Reuter, Till-Julius Brühl, Matthias Klein, Markus Hoffmann, Odile Filhol-Cochet, Irma Haben, Edgar Schmitt, Hajime Yurugi, Hans Christian Probst, Krishnaraj Rajalingam, Brigitte Boldyreff, Thierry Buchou, Iris Bellinghausen, Minka Breloer, Bastian Gerlitzki, Helmut Jonuleit, Andrea Tuettenberg, and Valérie Staudt
- Subjects
CD4-Positive T-Lymphocytes ,Male ,T cell ,Immunology ,Mice, Transgenic ,Receptors, Cell Surface ,chemical and pharmacologic phenomena ,Cell Growth Processes ,T-Lymphocytes, Regulatory ,Cell Line ,Mice ,Th2 Cells ,Immune system ,Hypersensitivity ,medicine ,Animals ,Humans ,Immunology and Allergy ,IL-2 receptor ,Casein Kinase II ,Mice, Inbred BALB C ,Chemistry ,Peripheral tolerance ,FOXP3 ,Cell Differentiation ,Forkhead Transcription Factors ,Dendritic Cells ,Acquired immune system ,Cell biology ,Mice, Inbred C57BL ,medicine.anatomical_structure ,Cell culture ,Interferon Regulatory Factors ,Leukocytes, Mononuclear ,IRF4 - Abstract
The quality of the adaptive immune response depends on the differentiation of distinct CD4(+) helper T cell subsets, and the magnitude of an immune response is controlled by CD4(+)Foxp3(+) regulatory T cells (Treg cells). However, how a tissue- and cell type-specific suppressor program of Treg cells is mechanistically orchestrated has remained largely unexplored. Through the use of Treg cell-specific gene targeting, we found that the suppression of allergic immune responses in the lungs mediated by T helper type 2 (TH2) cells was dependent on the activity of the protein kinase CK2. Genetic ablation of the β-subunit of CK2 specifically in Treg cells resulted in the proliferation of a hitherto-unexplored ILT3(+) Treg cell subpopulation that was unable to control the maturation of IRF4(+)PD-L2(+) dendritic cells required for the development of TH2 responses in vivo.
- Published
- 2015
26. Synthesis and biological evaluation of a novel MUC1 glycopeptide conjugate vaccine candidate comprising a 4’-deoxy-4’-fluoro-Thomsen–Friedenreich epitope
- Author
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Bastian Gerlitzki, Manuel Johannes, Maximilian Reindl, Anja Hoffmann-Röder, and Edgar Schmitt
- Subjects
Synthetic vaccine ,medicine.medical_treatment ,MUC1 ,Full Research Paper ,Epitope ,lcsh:QD241-441 ,Immune system ,Cancer immunotherapy ,lcsh:Organic chemistry ,Conjugate vaccine ,medicine ,skin and connective tissue diseases ,lcsh:Science ,neoplasms ,fluorinated carbohydrates ,cancer immunotherapy ,Chemistry ,Immunogenicity ,Organic Chemistry ,TACA ,digestive system diseases ,glycoconjugates ,Immunization ,Immunology ,Cancer research ,lcsh:Q ,Conjugate - Abstract
The development of selective anticancer vaccines that provide enhanced protection against tumor recurrence and metastasis has been the subject of intense research in the scientific community. The tumor-associated glycoprotein MUC1 represents a well-established target for cancer immunotherapy and has been used for the construction of various synthetic vaccine candidates. However, many of these vaccine prototypes suffer from an inherent low immunogenicity and are susceptible to rapid in vivo degradation. To overcome these drawbacks, novel fluorinated MUC1 glycopeptide-BSA/TTox conjugate vaccines have been prepared. Immunization of mice with the 4’F-TF-MUC1-TTox conjugate resulted in strong immune responses overriding the natural tolerance against MUC1 and producing selective IgG antibodies that are cross-reactive with native MUC1 epitopes on MCF-7 human cancer cells.
- Published
- 2015
27. CpG-Loaded Multifunctional Cationic Nanohydrogel Particles as Self-Adjuvanting Glycopeptide Antitumor Vaccines
- Author
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Natascha Stergiou, Horst Kunz, Björn Palitzsch, Lutz Nuhn, Edgar Schmitt, Sebastian Hartmann, Bastian Gerlitzki, Rudolf Zentel, and Markus Glaffig
- Subjects
medicine.medical_treatment ,Molecular Sequence Data ,Biomedical Engineering ,Pharmaceutical Science ,Enzyme-Linked Immunosorbent Assay ,medicine.disease_cause ,Cancer Vaccines ,Hydrogel, Polyethylene Glycol Dimethacrylate ,Epitope ,Biomaterials ,Adjuvants, Immunologic ,Cations ,medicine ,Animals ,Humans ,Amino Acid Sequence ,Receptor ,Mice, Inbred BALB C ,Oligonucleotide ,Toxin ,Chemistry ,Glycopeptides ,Glycopeptide ,Oligodeoxyribonucleotides ,CpG site ,Immunology ,Cancer research ,Nanoparticles ,Adjuvant ,Conjugate - Abstract
Self-adjuvanting antitumor vaccines by multifunctional cationic nanohydrogels loaded with CpG. A conjugate consisting of tumor-associated MUC1-glycopeptide B-cell epitope and tetanus toxin T-cell epitope P2 is linked to cationic nanogels. Oligonucleotide CpG complexation enhances toll-like receptor (TLR) stimulated T-cell proliferation and rapid immune activation. This co-delivery promotes induction of specific MUC1-antibodies binding to human breast tumor cells without external adjuvant.
- Published
- 2014
28. A Fully Synthetic Four-Component Antitumor Vaccine Consisting of a Mucin Glycopeptide Antigen Combined with Three Different T-Helper-Cell Epitopes
- Author
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Markus Glaffig, Horst Kunz, Sebastian Hartmann, Edgar Schmitt, Natascha Stergiou, and Björn Palitzsch
- Subjects
Molecular Structure ,Chemistry ,Immunogenicity ,Mucin-1 ,Antigen presentation ,Glycopeptides ,T-Lymphocytes, Helper-Inducer ,General Chemistry ,T helper cell ,Human leukocyte antigen ,Cancer Vaccines ,Virology ,Molecular biology ,Catalysis ,Epitope ,Glycopeptide ,Antigen-Antibody Reactions ,Epitopes ,medicine.anatomical_structure ,Antigen ,Antigens, Neoplasm ,medicine ,Humans ,MUC1 - Abstract
In a new concept of fully synthetic vaccines, the role of T-helper cells is emphasized. Here, a synthetic antitumor vaccine consisting of a diglycosylated tumor-associated MUC1 glycopeptide as the B-cell epitope was covalently cross-linked with three different T-helper-cell epitopes via squaric acid ligation of two linear (glyco)peptides. In mice this four-component vaccine administered without external immune-stimulating promoters elicit titers of MUC1-specific antibodies that were about eight times higher than those induced by a vaccine containing only one T-helper-cell epitope. The promising results indicate that multiple activation of different T-helper cells is useful for applications in which increased immunogenicity is required. In personalized medicine, in particular, this flexible construction of a vaccine can serve as a role model, for example, when T-helper-cell epitopes are needed that match human leukocyte antigens (HLA) in different patients.
- Published
- 2014
29. Eine vollsynthetische Vier-Komponenten-Antitumor-Vakzine mit einem MUC1-Glycopeptid und drei verschiedenen T-Helferzell- Epitopen
- Author
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Sebastian Hartmann, Natascha Stergiou, Horst Kunz, Edgar Schmitt, Markus Glaffig, and Björn Palitzsch
- Subjects
Chemistry ,General Medicine ,Molecular biology - Abstract
In einem neuen Konzept fur vollsynthetische Vakzine wird die Rolle von T-Helferzellen hervorgehoben. In einer solchen synthetischen Antitumor-Vakzine wurde ein zweifach glycosyliertes tumorassoziiertes MUC1-Glycopeptid als B-Zellepitop mit drei verschiedenen T-Helferzell-Epitopen durch Quadratsaurekonjugation zweier linearer (Glyco)Peptide kovalent verknupft. In Mausen loste die Impfung mit dieser Vier-Komponenten-Vakzine ohne zusatzliche Immunstimulantien etwa achtmal hohere MUC1-spezifische Antikorpertiter aus als eine Vakzine, die nur ein T-Helferzell-Epitop enthielt. Diese ermutigenden Ergebnisse zeigen, dass die gleichzeitige Aktivierung von T-Helferzellen verschiedener Spezifitat nutzlich fur Anwendungen ist, die eine gesteigerte Immunogenitat von Epitopen erfordern. Besonders in der personalisierten Medizin kann der flexible Aufbau der Vakzine als Vorbild dienen, wenn z. B. T-Helferzell-Epitope benotigt werden, die zum humanen Leukozytenantigen-Typ (HLA) verschiedener Patienten passen.
- Published
- 2014
30. Immunization with a Synthetic Human MUC1 Glycopeptide Vaccine against Tumor-Associated MUC1 Breaks Tolerance in Human MUC1 Transgenic Mice
- Author
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Helmut Jonuleit, Edgar Schmitt, Horst Kunz, Natascha Stergiou, and Markus Glaffig
- Subjects
0301 basic medicine ,Synthetic vaccine ,medicine.medical_treatment ,Breast Neoplasms ,Mice, Transgenic ,Biology ,01 natural sciences ,Biochemistry ,Cancer Vaccines ,03 medical and health sciences ,Immune system ,Antigen ,Cancer immunotherapy ,Drug Discovery ,medicine ,Tetanus Toxoid ,Animals ,Humans ,Antigens, Tumor-Associated, Carbohydrate ,General Pharmacology, Toxicology and Pharmaceutics ,Pharmacology ,Vaccines, Synthetic ,010405 organic chemistry ,Tetanus ,Organic Chemistry ,Mucin-1 ,Toxoid ,Immunotherapy ,medicine.disease ,Virology ,Peptide Fragments ,0104 chemical sciences ,Mice, Inbred C57BL ,030104 developmental biology ,Immunization ,Immunology ,MCF-7 Cells ,Molecular Medicine ,Female - Abstract
Breaking tolerance is crucial for effective tumor immunotherapy. We showed that vaccines containing tumor-associated human MUC1 glycopeptides induce strong humoral antitumor responses in mice. The question remained whether such vaccines work in humans, in systems where huMUC1 is a self-antigen. To clarify the question, mice transgenic in expressing huMUC1, mimicking the self-tolerant environment, and wild-type mice were vaccinated with a synthetic vaccine. This vaccine comprised STn and Tn antigens bound to a MUC1 tandem repeat peptide coupled to tetanus toxoid. The vaccine induced strong immune responses in wild-type and huMUC1-transgenic mice without auto-aggressive side effects. All antisera exhibited almost equivalent binding to human breast tumor cells. Similar increases of activated B-, CD4+ T-, and dendritic cells was found in the lymph nodes. The results demonstrate that tumor-associated huMUC1 glycopeptides coupled to tetanus toxoid are promising antitumor vaccines.
- Published
- 2017
31. Immunogenicity of a Fully Synthetic MUC1 Glycopeptide Antitumor Vaccine Enhanced by Poly(I:C) as a TLR3-Activating Adjuvant
- Author
-
Markus Glaffig, Edgar Schmitt, Horst Kunz, and Natascha Stergiou
- Subjects
0301 basic medicine ,medicine.medical_treatment ,chemical and pharmacologic phenomena ,Biochemistry ,Cancer Vaccines ,03 medical and health sciences ,Mice ,0302 clinical medicine ,Immune system ,Cancer immunotherapy ,Adjuvants, Immunologic ,Drug Discovery ,medicine ,Animals ,Humans ,General Pharmacology, Toxicology and Pharmaceutics ,MUC1 ,Pharmacology ,Vaccines, Synthetic ,biology ,Chemistry ,Immunogenicity ,Organic Chemistry ,Mucin-1 ,Glycopeptides ,Dendritic Cells ,Virology ,Glycopeptide ,Toll-Like Receptor 3 ,030104 developmental biology ,Poly I-C ,030220 oncology & carcinogenesis ,TLR3 ,biology.protein ,Molecular Medicine ,Antibody ,Adjuvant - Abstract
Fully synthetic MUC1 glycopeptide antitumor vaccines have a precisely specified structure and induce a targeted immune response without suppression of the immune response when using an immunogenic carrier protein. However, tumor-associated aberrantly glycosylated MUC1 glycopeptides are endogenous structures, “self-antigens”, that exhibit only low immunogenicity. To overcome this obstacle, a fully synthetic MUC1 glycopeptide antitumor vaccine was combined with poly(inosinic acid:cytidylic acid), poly(I:C), as a structurally defined Toll-like receptor 3 (TLR3)-activating adjuvant. This vaccine preparation elicited extraordinary titers of IgG antibodies which strongly bound human breast cancer cells expressing tumor-associated MUC1. Beside the humoral response, the poly(I:C) glycopeptide vaccine induced a pro-inflammatory environment, very important to overcome the immune-suppressive mechanisms, and elicited a strong cellular immune response crucial for tumor elimination.
- Published
- 2017
32. Reversible Covalent and Supramolecular Functionalization of Water-Soluble Gold(I) Complexes
- Author
-
Daniel Spitzer, Edgar Schmitt, Bart Jan Ravoo, Natascha Stergiou, Vanessa Lewe, Benedict Kemper, Maximilian von Gröning, Dieter Schollmeyer, Pol Besenius, and Tobias Otremba
- Subjects
010405 organic chemistry ,Organic Chemistry ,Supramolecular chemistry ,Dynamic covalent chemistry ,General Chemistry ,010402 general chemistry ,01 natural sciences ,Catalysis ,Cycloaddition ,0104 chemical sciences ,chemistry.chemical_compound ,chemistry ,Covalent bond ,Biotinylation ,Polymer chemistry ,Maleimide ,Carbene ,Conjugate - Abstract
The ligation of gold(I) metalloamphiphiles with biomolecules is reported, using water-soluble AuI -N-alkynyl substituted maleimide complexes. For this purpose, two different polar ligands were applied: 1) a neutral, dendritic tetraethylene glycol-functionalized phosphane and 2) a charged, sulfonated N-heterocyclic carbene (NHC). The retro Diels-Alder reaction of a furan-protected maleimide gold(I) complex, followed by cycloaddition with a diene-functionalized biotin under mild conditions leads to a novel gold(I) metalloamphiphile. The strong streptavidin-biotin binding affinity in buffered aqueous solution of the resulting biotin alkynyl gold(I) phosphane conjugate remains intact. The cytotoxicity of the biotinylated gold(I) complex against a T47D human breast cancer cell line is higher than for cisplatin.
- Published
- 2017
33. Messenger RNA Sequencing of Rare Cell Populations in the Lung and Lung-Draining Lymph Nodes
- Author
-
Edgar Schmitt, Alexander Ulges, Tobias Bopp, and Matthias Klein
- Subjects
Transcriptome ,Genetics ,Gene expression profiling ,medicine.anatomical_structure ,Single-cell analysis ,T cell ,Gene expression ,Cell ,medicine ,RNA ,Genomic library ,Biology - Abstract
Next-generation sequencing (NGS) techniques provide unique prospects for in-depth transcriptome analyses. Nevertheless, the emerging and still growing knowledge about the large diversity and heterogeneity of cells that participate in immunological responses in a tissue- and micromilieu-specific manner calls for advanced isolation and sequencing methods for the accurate quantification of gene expression in small cell populations and even individual cells from any organ or tissue. One of the major limitations in performing transcriptome analyses of rare cell populations was and still is quality and quantity of RNA that often limits analyses of complex mixtures of immune cell populations. Here, we describe a protocol to isolate rare T cell populations from the lung and in particular the subsequent methods to isolate high-grade RNA in order to perform NGS-based transcriptome analyses.
- Published
- 2017
34. Microarray analysis of antibodies induced with synthetic antitumor vaccines : specificity against diverse mucin core structures
- Author
-
Manuel Schorlemer, Jia Liu, Hui Cai, Björn Palitzsch, Mengji Lu, Edgar Schmitt, Natascha Stergiou, Horst Kunz, Ulrika Westerlind, Christian Pett, and Sebastian Hartmann
- Subjects
Antigenicity ,Glycosylation ,Antibody microarray ,Protein Array Analysis ,Medizin ,010402 general chemistry ,01 natural sciences ,Cancer Vaccines ,Catalysis ,Epitope ,chemistry.chemical_compound ,Mice ,Antigen ,Polysaccharides ,Neoplasms ,Animals ,Humans ,MUC1 ,Vaccines, Synthetic ,biology ,010405 organic chemistry ,Organic Chemistry ,Mucin-1 ,Glycopeptides ,General Chemistry ,Combinatorial chemistry ,0104 chemical sciences ,Immunity, Humoral ,chemistry ,Biochemistry ,Polyclonal antibodies ,Antibody Formation ,biology.protein ,Antibody - Abstract
Glycoprotein research is pivotal for vaccine development and biomarker discovery. Many successful methodologies for reliably increasing the antigenicity toward tumor-associated glycopeptide structures have been reported. Deeper insights into the quality and specificity of the raised polyclonal, humoral reactions are often not addressed, despite the fact that an immunological memory, which produces antibodies with cross-reactivity to epitopes exposed on healthy cells, may cause autoimmune diseases. In the current work, three MUC1 antitumor vaccine candidates conjugated with different immune stimulants are evaluated immunologically. For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. This is one of the most extensive glycopeptide libraries ever made through total synthesis. On tumor cells, the core 2 β-1,6-N-acetylglucosaminyltransferase-1 (C2GlcNAcT-1) is down-regulated, resulting in lower amounts of the branched core 2 structures, which favor formation of linear core 1 or core 3 structures, and in particular, truncated tumor-associated antigen structures. The core 2 structures are commonly found on healthy cells and the elucidation of antibody cross-reactivity to such epitopes may predict the tumor-selectivity and safety of synthetic vaccines. With the extended mucin core structures in hand, antibody cross-reactivity toward the branched core 2 glycopeptide epitopes is explored. It is observed that the induced antibodies recognize MUC1 peptides with very high glycosylation site specificity. The nature of the antibody response is characteristically different for antibodies directed to glycosylation sites in either the immune-dominant PDTR or the GSTA domain. All antibody sera show high reactivity to the tumor-associated saccharide structures on MUC1. Extensive glycosylation with branched core 2 structures, typically found on healthy cells, abolishes antibody recognition of the antisera and suggests that all vaccine conjugates preferentially induce a tumor-specific humoral immune response.
- Published
- 2017
35. Donor and host B cell-derived IL-10 contributes to suppression of graft-versus-host disease
- Author
-
Hansjörg Schild, Tobias Bopp, Andreas Kreft, Pamela Stein, Axel Roers, Edgar Schmitt, Marc Weber, Simon Fillatreau, Markus P. Radsak, Berenice M. Rudolph, and Steve Prüfer
- Subjects
biology ,medicine.medical_treatment ,Immunology ,chemical and pharmacologic phenomena ,Spleen ,medicine.disease ,Interleukin 10 ,surgical procedures, operative ,medicine.anatomical_structure ,Graft-versus-host disease ,Cytokine ,immune system diseases ,CD1D ,biology.protein ,medicine ,Immunology and Allergy ,CD5 ,Mode of action ,B cell - Abstract
Graft-versus-host disease (GvHD) is a frequent life-threatening complication following allogeneic HSC transplantation (HSCT). IL-10 is a regulatory cytokine with important roles during GvHD, yet its relevant sources, and mode of action, remain incompletely defined in this disease. Using IL-10-deficient donor or host mice (BALB/c or C57BL/6, respectively) in a MHC-mismatched model for acute GvHD, we found a strongly aggravated course of the disease with increased mortality when either donor or host cells could not produce this cytokine. A lack of IL-10 resulted in increased allogeneic T-cell responses and enhanced activation of host DCs in spleen and MLNs. Remarkably, IL-10 was prominently produced by host- and donor-derived CD5(int) CD1d(int) TIM-1(int) B cells in this disease, and consistent with this, allogeneic HSCT resulted in exacerbated GvHD when mice lacking IL-10 expression in B cells were used as donor or host, compared with controls. Taken together, this study demonstrates that host and donor B cell-derived IL-10 provides a unique mechanism of suppression of acute GvHD, and suggests that DCs are the targets of this B cell-mediated suppressive effect. These findings open novel therapeutic possibilities based on the use of B cells to increase the feasibility of allogeneic HSCT.
- Published
- 2014
36. Mit Glycopeptid-Antigenen und T-Zell-Epitopen verknüpfte wasserlösliche Polymere als potenzielle Antitumor-Vakzine
- Author
-
Edgar Schmitt, Björn Palitzsch, Horst Kunz, Sebastian Hartmann, Rudolf Zentel, Lutz Nuhn, and Bastian Gerlitzki
- Subjects
Chemistry ,General Medicine - Published
- 2013
37. Water-Soluble Polymers Coupled with Glycopeptide Antigens and T-Cell Epitopes as Potential Antitumor Vaccines
- Author
-
Lutz Nuhn, Bastian Gerlitzki, Björn Palitzsch, Sebastian Hartmann, Rudolf Zentel, Edgar Schmitt, and Horst Kunz
- Subjects
Synthetic vaccine ,Molecular Sequence Data ,Epitopes, T-Lymphocyte ,Cancer Vaccines ,Catalysis ,Epitope ,Mice ,chemistry.chemical_compound ,Polymethacrylic Acids ,Antigen ,Animals ,Humans ,Methacrylamide ,Amino Acid Sequence ,MUC1 ,Vaccines, Synthetic ,biology ,Mucin-1 ,Glycopeptides ,Toxoid ,Water ,T-Lymphocytes, Helper-Inducer ,General Chemistry ,Molecular biology ,Glycopeptide ,Solubility ,Biochemistry ,chemistry ,MCF-7 Cells ,biology.protein ,Antibody - Abstract
Highly decorated: Tumor-associated MUC1 glycopeptide and tetanus toxoid T-cell epitope P2 can be attached to water-soluble poly(N-(2-hydroxypropyl)methacrylamide) carriers by orthogonal ligation techniques. Fully synthetic vaccine A with additional nanostructure-promoting domains induced antibodies that exhibit high affinity to tumor cells.
- Published
- 2013
38. Mast Cell–deficient KitW-sh 'Sash' Mutant Mice Display Aberrant Myelopoiesis Leading to the Accumulation of Splenocytes That Act as Myeloid-Derived Suppressor Cells
- Author
-
Andrea Schüler, Pamela Friedrich, Edgar Schmitt, Fatma Döner, Stefan Tenzer, Ute Distler, Anastasija Michel, Marc Becker, Tobias Bopp, Markus P. Radsak, Markus Hoffmann, Jörg Kuharev, Michael Stassen, Manfred Relle, Thorsten B. Feyerabend, Hansjörg Schild, and Hans Reimer Rodewald
- Subjects
Cell type ,Myeloid ,T cell ,Immunology ,Biology ,Immunophenotyping ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Neoplasms ,medicine ,Animals ,Antigens, Ly ,Immunology and Allergy ,Myeloid Cells ,Mast Cells ,Progenitor cell ,030304 developmental biology ,Mice, Knockout ,Myelopoiesis ,0303 health sciences ,CD11b Antigen ,Mast cell ,Adoptive Transfer ,3. Good health ,Cell biology ,Proto-Oncogene Proteins c-kit ,Haematopoiesis ,medicine.anatomical_structure ,Hematopoiesis, Extramedullary ,Mutation ,Myeloid-derived Suppressor Cell ,Female ,Neoplasm Transplantation ,Spleen ,030215 immunology - Abstract
Mast cell-deficient KitW-sh “sash” mice are widely used to investigate mast cell functions. However, mutations of c-Kit also affect additional cells of hematopoietic and nonimmune origin. In this study, we demonstrate that KitW-sh causes aberrant extramedullary myelopoiesis characterized by the expansion of immature lineage-negative cells, common myeloid progenitors, and granulocyte/macrophage progenitors in the spleen. A consistent feature shared by these cell types is the reduced expression of c-Kit. Populations expressing intermediate and high levels of Ly6G, a component of the myeloid differentiation Ag Gr-1, are also highly expanded in the spleen of sash mice. These cells are able to suppress T cell responses in vitro and phenotypically and functionally resemble myeloid-derived suppressor cells (MDSC). MDSC typically accumulate in tumor-bearing hosts and are able to dampen immune responses. Consequently, transfer of MDSC from naive sash mice into line 1 alveolar cell carcinoma tumor-bearing wild-type littermates leads to enhanced tumor progression. However, although it can also be observed in sash mice, accelerated growth of transplanted line 1 alveolar cell carcinoma tumors is a mast cell–independent phenomenon. Thus, the KitW-sh mutation broadly affects key steps in myelopoiesis that may have an impact on mast cell research.
- Published
- 2013
39. NFATc1 Induction in Peripheral T and B Lymphocytes
- Author
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Rene Rost, Matthias Hock, Duong Anh Thuy Pham, Martin Vaeth, Ronald Rudolf, Dimitri Tyrsin, Khalid Muhammad, Edgar Schmitt, Edgar Serfling, Amiya K. Patra, Friederike Berberich-Siebelt, Sergei Chuvpilo, Andris Avots, Tobias Bopp, Tobias Pusch, and Stefan Klein-Hessling
- Subjects
Lipopolysaccharides ,Gene isoform ,Chromosomes, Artificial, Bacterial ,Programmed cell death ,Transgene ,Green Fluorescent Proteins ,Immunology ,Gene Expression ,Mice, Transgenic ,Stimulation ,Immune receptor ,Biology ,Lymphocyte Activation ,T-Lymphocytes, Regulatory ,Antibodies ,Mice ,Th2 Cells ,Genes, Reporter ,Transforming Growth Factor beta ,Animals ,Protein Isoforms ,Immunology and Allergy ,Promoter Regions, Genetic ,Transcription factor ,Cell Proliferation ,B-Lymphocytes ,NFATC Transcription Factors ,integumentary system ,NF-kappa B ,CD28 ,NFAT ,Th1 Cells ,Molecular biology ,Mice, Inbred C57BL ,Th17 Cells - Abstract
NFAT transcription factors control the proliferation and survival of peripheral lymphocytes. We have reported previously that the short isoform NFATc1/αA whose generation is induced by immune receptor stimulation supports the proliferation and inhibits the activation-induced cell death of peripheral T and B cells. We will show in this study that in novel bacterial artificial chromosome transgenic mice that express EGFP under the control of entire Nfatc1 locus the Nfatc1/Egfp transgene is expressed as early as in double-negative thymocytes and in nonstimulated peripheral T and B cells. Upon immune receptor stimulation, Nfatc1/Egfp expression is elevated in B, Th1, and Th2 cells, but only weakly in T regulatory, Th9, and Th17 cells in vitro whose generation is affected by TGFβ. In naive lymphocytes, persistent immune receptor signals led to a 3–5 increase in NFATc1/αA RNA levels during primary and secondary stimulation, but a much stronger induction was observed at the protein level. Whereas anti-CD3+CD28 stimulation of primary T cells induces both NFATc1/αA and their proliferation and survival, anti-IgM stimulation of B cells induces NFATc1/αA and proliferation, but activation-induced cell death after 3-d incubation in vitro. The anti-IgM–mediated activation-induced cell death induction of B cells in vitro is suppressed by anti-CD40–, LPS-, and CpG-mediated signals. In addition to inducing NF-κB factors, together with anti-IgM, these signals also support the generation of NFATc1/αA. According to these data and the architecture of its promoter region, the Nfatc1 gene resembles a primary response gene whose induction is affected at the posttranscriptional level.
- Published
- 2013
40. The extent of HLA-DR expression on HLA-DR+Tregs allows the identification of patients with clinically relevant borderline rejection
- Author
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Edgar Schmitt, Martin Zeier, Stefan Meuer, Luis E. Becker, Nicole Seissler, Sebastian Markus Schaefer, Rüdiger Waldherr, Friederike Hug, Andrea Steinborn, Matthias Schaier, and Claudia Sommerer
- Subjects
Adult ,Graft Rejection ,Male ,Regulatory T cell ,Risk Assessment ,Sensitivity and Specificity ,T-Lymphocytes, Regulatory ,Flow cytometry ,Cohort Studies ,Pathogenesis ,Young Adult ,Predictive Value of Tests ,Reference Values ,Biopsy ,medicine ,HLA-DR ,Humans ,Survival rate ,Aged ,Subclinical infection ,Transplantation ,medicine.diagnostic_test ,business.industry ,Biopsy, Needle ,Forkhead Transcription Factors ,HLA-DR Antigens ,Middle Aged ,Flow Cytometry ,Immunohistochemistry ,Kidney Transplantation ,Survival Rate ,Treatment Outcome ,medicine.anatomical_structure ,ROC Curve ,Case-Control Studies ,Immunology ,Linear Models ,Kidney Failure, Chronic ,Female ,business ,Biomarkers - Abstract
Regulatory T cells (Tregs) were shown to be involved into the pathogenesis of acute rejection after transplantation. The suppressive activity of the total regulatory T cell pool depends on its percentage of highly suppressive HLA-DR(+) -Treg cells. Therefore, both the suppressive activity of the total Treg pool and the extent of HLA-DR expression of HLA-DR(+) -Tregs (MFI HLA-DR) were estimated in non transplanted volunteers, patients with end-stage renal failure (ESRF), healthy renal transplant patients with suspicion on rejection, due to sole histological Bord-R or sole acute renal failure (ARF), and patients with clinically relevant borderline rejection (Bord-R and ARF). Compared to patients with only Bord-R or only ARF, the suppressive activity of the total Treg cell pool was exclusively reduced in patients with clinically relevant Bord-R. In parallel, the HLA-DR MFI of the DR(+) -Treg subset was significantly decreased in these patients, due to a significantly lower proportion of DR(high+) -Tregs, which were shown to have the highest suppressive capacity within the total Treg pool. Our findings clearly demonstrate that the determination of the HLA-DR MFI of the HLA-DR(+) -Treg subset allows a highly sensitive, specific and non-invasive discrimination between patients with clinically relevant Bord-R (Bord and ARF) and patients with subclinical rejection or other causes of transplant failure.
- Published
- 2013
41. Intrahepatic myeloid-cell aggregates enable local proliferation of CD8+T cells and successful immunotherapy against chronic viral liver infection
- Author
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Ulrike Protzer, Li-Rung Huang, Florian Reisinger, Craig N. Jenne, Hans-Peter Dienes, Zeinab Abdullah, Paul Kubes, Nico van Rooijen, Natalio Garbi, Margarete Odenthal, Edgar Schmitt, Michael Croft, Frank A. Schildberg, Percy A. Knolle, Dirk Wohlleber, Mathias Heikenwalder, Ru-Lin Cheng, Christian Kurts, Molecular cell biology and Immunology, and CCA - Innovative therapy
- Subjects
T cell ,medicine.medical_treatment ,Immunology ,Population ,Green Fluorescent Proteins ,Mice, Transgenic ,Biology ,CD8-Positive T-Lymphocytes ,Lymphocytic Choriomeningitis ,Mice ,medicine ,Immunology and Allergy ,Cytotoxic T cell ,Animals ,Lymphocytic choriomeningitis virus ,Myeloid Cells ,education ,Cell Proliferation ,Mice, Knockout ,education.field_of_study ,Liver infection ,CD11b Antigen ,Microscopy, Confocal ,Liver Diseases ,Immunotherapy ,Receptors, OX40 ,Flow Cytometry ,Mice, Inbred C57BL ,CTL ,Chronic infection ,medicine.anatomical_structure ,Animals, Newborn ,Liver ,Toll-Like Receptor 9 ,Chronic Disease ,Host-Pathogen Interactions ,CD8 ,Signal Transduction ,T-Lymphocytes, Cytotoxic - Abstract
Chronic infection is difficult to overcome because of exhaustion or depletion of cytotoxic effector CD8(+) T cells (cytotoxic T lymphoytes (CTLs)). Here we report that signaling via Toll-like receptors (TLRs) induced intrahepatic aggregates of myeloid cells that enabled the population expansion of CTLs (iMATEs: 'intrahepatic myeloid-cell aggregates for T cell population expansion') without causing immunopathology. In the liver, CTL proliferation was restricted to iMATEs that were composed of inflammatory monocyte-derived CD11b(+) cells. Signaling via tumor-necrosis factor (TNF) caused iMATE formation that facilitated costimulation dependent on the receptor OX40 for expansion of the CTL population. The iMATEs arose during acute viral infection but were absent during chronic viral infection, yet they were still induced by TLR signaling. Such hepatic expansion of the CTL population controlled chronic viral infection of the liver after vaccination with DNA. Thus, iMATEs are dynamic structures that overcome regulatory cues that limit the population expansion of CTLs during chronic infection and can be used in new therapeutic vaccination strategies.
- Published
- 2013
42. Methylprednisolone treatment increases the proportion of the highly suppressive HLA-DR+-Treg-cells in transplanted patients
- Author
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Andrea Steinborn, Claudia Sommerer, Martin Zeier, Nicole Seissler, Friederike Hug, Matthias Schaier, and Edgar Schmitt
- Subjects
Graft Rejection ,Immunology ,Population ,Cell ,chemical and pharmacologic phenomena ,Adaptive Immunity ,Methylprednisolone ,T-Lymphocytes, Regulatory ,Bolus (medicine) ,Immune system ,Transplantation Immunology ,Biopsy ,medicine ,HLA-DR ,Humans ,Immunology and Allergy ,education ,Glucocorticoids ,Transplantation ,education.field_of_study ,Innate immune system ,medicine.diagnostic_test ,business.industry ,hemic and immune systems ,HLA-DR Antigens ,Kidney Transplantation ,Immunity, Innate ,medicine.anatomical_structure ,business ,Immunosuppressive Agents ,medicine.drug - Abstract
Methylprednisolone is widely used to improve immune suppression in transplanted patients threatened by acute rejection. Recently, we showed that the suppressive activity of a Treg cell population depends decisively on their percentage of highly suppressive HLA-DR(high+)-Treg cells, which are strongly reduced in rejecting transplant patients. In order to examine whether the composition of the total CD4(+)CD127(low+/-)FoxP3(+)-Treg cell pool with different Treg-subsets (DR(high+)CD45RA(-)-Tregs, DR(low+)CD45RA(-)-Tregs, DR(-)CD45RA(-)-Tregs, DR(-)CD45RA(+)-Tregs) is affected by methylprednisolone bolus therapy we compared the percentages of these four different Treg cell subsets in transplant patients with biopsy proven rejection before and after steroid bolus therapy (n=23). In patients treated with steroid bolus therapy, the percentage of the naïve DR(-)CD45RA(+)-Tregs was significantly decreased, whereas the percentage of the DR(+)CD45RA(-)-Tregs was significantly increased. By that, the strongest increase was detected for the most suppressive DR(high+)CD45RA(-)-Tregs. However, these effects were only temporarily and closely associated to the duration of the bolus therapy. Our results suggest that besides various anti-inflammatory effects on cells of the adaptive and innate immune system, methylprednisolone also has the capacity to enhance the suppressive activity of the total Treg cell pool by increasing its percentage of highly differentiated and highly suppressive DR(high+)CD45RA(-)-Tregs.
- Published
- 2012
43. Discovery and initial characterization of Th9 cells: the early years
- Author
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Tobias Bopp and Edgar Schmitt
- Subjects
0301 basic medicine ,Adoptive cell transfer ,Immunology ,Biology ,03 medical and health sciences ,Interleukin 21 ,Mice ,T-Lymphocyte Subsets ,medicine ,Immunology and Allergy ,Cytotoxic T cell ,Animals ,Humans ,IL-2 receptor ,Cloning, Molecular ,Peripheral neuritis ,Receptors, Interleukin-9 ,Research ,Interleukin-9 ,T helper cell ,T-Lymphocytes, Helper-Inducer ,History, 20th Century ,030104 developmental biology ,Interleukin 31 ,medicine.anatomical_structure ,Gene Expression Regulation ,Interleukin 13 ,Cytokines - Abstract
The launch of the Th1/Th2 concept represented a decisive breakthrough concerning our understanding of how very diverse immune reactions can be regulated by functionally different T helper subpopulations via the secretion of different panels of cytokines. In this context, IL-9 was identified to be produced by T helper cell lines in addition to Th2 cytokines IL-4 and IL-5. Detailed analyses revealed that IL-9 production of mouse CD4+ T helper cells was dependent on a combination of IL-2, IL-4, and TGF-β. Roughly a decade later, it was found that TGF-β can also induce the development of CD4+ Treg cells. This finding engendered a series of studies on the central role of TGF-β for cytokine-mediated T helper cell differentiation which elucidated that IL-4 curbed the Treg cell-promoting effect of TGF-β while TGF-β impaired the Th2-promoting capacity of IL-4. Instead, TGF-β in combination with IL-4 induced the development of CD4+ T helper cells that preferentially produced IL-9 and that were different from Th2 cells which originally were thought to be the main source of IL-9. In addition, adoptive transfer of such IL-9-producing CD4+ T helper cells was shown to cause the development of colitis and peripheral neuritis. Hence, the unique cytokine expression pattern in combination with the inflammatory in vivo phenotype led to the designation of Th9 cells as a new CD4+ T helper subpopulation.
- Published
- 2016
44. Interleukin 9
- Author
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Edgar Schmitt, Matthias Klein, and Tobias Bopp
- Published
- 2016
45. Context- and Tissue-Specific Regulation of Immunity and Tolerance by Regulatory T Cells
- Author
-
Edgar Schmitt, Tobias Bopp, Christoph Becker, and Alexander Ulges
- Subjects
0301 basic medicine ,Regulatory T cell ,FOXP3 ,chemical and pharmacologic phenomena ,Context (language use) ,Inflammation ,Biology ,03 medical and health sciences ,030104 developmental biology ,medicine.anatomical_structure ,Immune system ,Antigen ,Immunity ,Immunopathology ,Immunology ,medicine ,medicine.symptom - Abstract
The immune system has evolved to defend the organism against an almost infinite number of pathogens in a locally confined and antigen-specific manner while at the same time preserving tolerance to harmless antigens and self. Regulatory T (Treg) cells essentially contribute to an immunoregulatory network preventing excessive immune responses and immunopathology. There is emerging evidence that Treg cells not only operate in secondary lymphoid tissue but also regulate immune responses directly at the site of inflammation. Hence, the classification of Treg cells might need to be further extended by Treg cell subsets that are functionally and phenotypically polarized by their residency. In this review, we discuss recent findings on these tissue-resident Treg cell subsets and how these cells may operate in a tissue- and context-dependent manner.
- Published
- 2016
46. Combined B, T and NK Cell Deficiency Accelerates Atherosclerosis in BALB/c Mice
- Author
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Leonard D. Shultz, Kurt Reifenberg, Kerstin Winter, Eva Karoline Pross, Fei Cheng, Edgar Schmitt, Laura Twardowski, Jianglin Fan, Karl J. Lackner, Michael Torzewski, and Antje Canisius
- Subjects
0301 basic medicine ,T-Lymphocytes ,lcsh:Medicine ,NK cells ,Adaptive Immunity ,Biochemistry ,Vascular Medicine ,Mice ,chemistry.chemical_compound ,Cellular types ,Receptor ,lcsh:Science ,Immunodeficiency ,Mice, Knockout ,B-Lymphocytes ,Mice, Inbred BALB C ,Multidisciplinary ,biology ,T Cells ,Immune cells ,Acquired immune system ,Lipids ,Plaque, Atherosclerotic ,Killer Cells, Natural ,Cholesterol ,Phenotype ,White blood cells ,Female ,lipids (amino acids, peptides, and proteins) ,Research Article ,Cell biology ,Blood cells ,Lipoproteins ,Immunology ,Research and Analysis Methods ,BALB/c ,Immune Deficiency ,03 medical and health sciences ,Immune system ,medicine ,Animals ,Immunohistochemistry Techniques ,Triglycerides ,Medicine and health sciences ,Biology and life sciences ,Macrophages ,lcsh:R ,Immunologic Deficiency Syndromes ,Wild type ,Proteins ,Atherosclerosis ,biology.organism_classification ,medicine.disease ,Molecular biology ,Histochemistry and Cytochemistry Techniques ,030104 developmental biology ,Animal cells ,Receptors, LDL ,chemistry ,Immune System ,Mutation ,LDL receptor ,Immunologic Techniques ,Clinical Immunology ,lcsh:Q ,Clinical Medicine - Abstract
This study focused on the unique properties of both the Ldlr knockout defect (closely mimicking the human situation) and the BALB/c (C) inbred mouse strain (Th-2 slanted immune response). We generated two immunodeficient strains with severe combined B- and T-cell immunodeficiency with or without a complete lack of natural killer cells to revisit the role of adaptive immune responses on atherogenesis. C-Ldlr-/- Rag1-/- mice, which show severe combined B- and T-cell immunodeficiency and C-Ldlr-/- Rag1-/- Il2rg-/- mice, which combine the T- and B-cell defect with a complete lack of natural killer cells and inactivation of multiple cytokine signalling pathways were fed an atherogenic Western type diet (WTD). Both B6-Ldlr-/- and C-Ldlr-/- immunocompetent mice were used as controls. Body weights and serum cholesterol levels of both immunodeficient strains were significantly increased compared to C-Ldlr-/- controls, except for cholesterol levels of C-Ldlr-/- Rag1-/- double mutants after 12 weeks on the WTD. Quantification of the aortic sinus plaque area revealed that both strains of immunodeficient mice developed significantly more atherosclerosis compared to C-Ldlr-/- controls after 24 weeks on the WTD. Increased atherosclerotic lesion development in C-Ldlr-/- Rag1-/- Il2rg-/- triple mutants was associated with significantly increased numbers of macrophages and significantly decreased numbers of smooth muscle cells compared to both C-Ldlr-/- wild type and C-Ldlr-/- Rag1-/- double mutants pointing to a plaque destabilizing effect of NK cell loss. Collectively, the present study reveals a previously unappreciated complexity with regard to the impact of lymphocytes on lipoprotein metabolism and the role of lymphocyte subsets in plaque composition.
- Published
- 2016
47. Term and preterm labor: decreased suppressive activity and changes in composition of the regulatory T‐cell pool
- Author
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Edgar Schmitt, Diana Radnai, Andrea Steinborn, Linda Schober, Christof Sohn, and Karsten Mahnke
- Subjects
medicine.medical_specialty ,Term Birth ,Regulatory T cell ,Immunology ,chemical and pharmacologic phenomena ,T-Lymphocytes, Regulatory ,Immunophenotyping ,Obstetric Labor, Premature ,Antigens, CD ,Pregnancy ,T-Lymphocyte Subsets ,Internal medicine ,Immune Tolerance ,medicine ,Humans ,Immunology and Allergy ,IL-2 receptor ,Interleukin-7 receptor ,Cells, Cultured ,Fetus ,business.industry ,Infant, Newborn ,FOXP3 ,Forkhead Transcription Factors ,hemic and immune systems ,HLA-DR Antigens ,Cell Biology ,medicine.disease ,Surgery ,Tolerance induction ,Endocrinology ,medicine.anatomical_structure ,Female ,Composition (visual arts) ,business - Abstract
Regulatory T cells (Tregs) exert a key role in tolerance induction to the semi-allogeneic fetus. Currently, it is not known whether immunological rejection processes are involved in the induction of normal term or irresistible preterm labor. In this study, we examined whether there were differences in the percentage of the total CD4 þ CD127 low þ/ � CD25 þ FoxP3 þ -Treg-cell pool, its suppressive activity and its composition with distinct Treg subsets (HLA-DR low þ -, HLA-DR high þ -, HLA-DR � - and naive CD45RA þ -Tregs) between preterm and term laboring women. We found that its percentage was decreased neither in term nor in preterm laboring women. Its suppressive activity was strongly diminished in preterm laboring women and to a lesser extent in spontaneously term laboring women. During the normal course of pregnancy, its composition changed in such a way that the percentage of naive CD45RA þ -Tregs increased while the percentage of the highly suppressive HLA-DR low þ - and HLA-DR high þ Tregs decreased significantly until term. With the onset of spontaneous term labor this phenomenon was reversed and reached significant values postpartum. In addition, we confirmed that both the decreased percentage of HLA-DR þ -Tregs within the total Treg-cell pool and their decreased level of HLA-DR expression (depending on the percentage of HLA-DR low þ - and HLA-DR high þ Tregs) had a reducing effect on the suppressive activity of the total Treg cell pool in preterm laboring women. However, spontaneous term delivery was associated with increasing percentages of HLA-DR þ -Tregs and increasing HLA-DR expression of this Treg subset. Therefore, it becomes apparent that the mechanisms inducing term or preterm labor may be completely different.
- Published
- 2012
48. Repression of Cyclic Adenosine Monophosphate Upregulation Disarms and Expands Human Regulatory T Cells
- Author
-
Ria Baumgrass, Stephan Grabbe, Christian Becker, Tobias Scheel, Martin Vaeth, Edgar Schmitt, Tobias Bopp, Matthias Klein, and Friederike Berberich-Siebelt
- Subjects
medicine.medical_specialty ,Regulatory T cell ,Immunology ,chemical and pharmacologic phenomena ,Biology ,T-Lymphocytes, Regulatory ,Mice ,chemistry.chemical_compound ,Internal medicine ,Cyclic AMP ,medicine ,Animals ,Humans ,Immunology and Allergy ,Cyclic adenosine monophosphate ,Psychological repression ,Cell Proliferation ,Clonal Anergy ,NFATC Transcription Factors ,Clonal anergy ,Phosphodiesterase ,hemic and immune systems ,Up-Regulation ,Cell biology ,Endocrinology ,medicine.anatomical_structure ,chemistry ,Humanized mouse ,cAMP-dependent pathway ,Cyclase activity - Abstract
The main molecular mechanism of human regulatory T cell (Treg)-mediated suppression has not been elucidated. We show in this study that cAMP represents a key regulator of human Treg function. Repression of cAMP production by inhibition of adenylate cyclase activity or augmentation of cAMP degradation through ectopic expression of a cAMP-degrading phosphodiesterase greatly reduces the suppressive activity of human Treg in vitro and in a humanized mouse model in vivo. Notably, cAMP repression additionally abrogates the anergic state of human Treg, accompanied by nuclear translocation of NFATc1 and induction of its short isoform NFATc1/αA. Treg expanded under cAMP repression, however, do not convert into effector T cells and regain their anergic state and suppressive activity upon proliferation. Together, these findings reveal the cAMP pathway as an attractive target for clinical intervention with Treg function.
- Published
- 2012
49. From interleukin-9 to T helper 9 cells
- Author
-
Michael Stassen, Tobias Bopp, and Edgar Schmitt
- Subjects
General Neuroscience ,ZAP70 ,T helper cell ,Biology ,General Biochemistry, Genetics and Molecular Biology ,Interleukin 21 ,medicine.anatomical_structure ,History and Philosophy of Science ,Immunology ,medicine ,Cytotoxic T cell ,Interleukin 9 ,IL-2 receptor ,Antigen-presenting cell ,Interleukin 3 - Abstract
Interleukin-9 (IL-9), cloned more than 20 years ago, was initially thought to be a Th2-specific cytokine. This assumption was initially confirmed by functional analyses showing that both IL-9 and Th2 cells play an important role in the pathogenesis of asthma, IgE class switch recombination, and resolution of parasitic infections. However, recently it was shown that IL-9-producing CD4(+) T cells represent the discrete T helper subset Th9 cells. Herein, we will review the cytokines and transcription factors known to promote the development of Th9 cells and their potential functional properties in relation to the biological activities of IL-9. In addition, we will discuss how Th9 cells are related to Th2, Th17, and T(reg) cells, as both an alternative source of IL-9 and in view of the fact that plasticity of CD4(+) T cell differentiation is currently a strong matter of debate in immunologic research.
- Published
- 2012
50. A Synthetic Glycopeptide Vaccine for the Induction of a Monoclonal Antibody that Differentiates between Normal and Tumor Mammary Cells and Enables the Diagnosis of Human Pancreatic Cancer
- Author
-
Natascha Stergiou, Nicole Teusch, Horst Kunz, Björn Palitzsch, Edgar Schmitt, Sebastian Hartmann, Nikola Gaidzik, Bastian Gerlitzki, Peer Flemming, and Sonja Stahn
- Subjects
medicine.drug_class ,medicine.medical_treatment ,Tumor M2-PK ,Breast Neoplasms ,Biology ,010402 general chemistry ,Monoclonal antibody ,01 natural sciences ,Cancer Vaccines ,Catalysis ,Cancer immunotherapy ,Antigen ,Pancreatic tumor ,Pancreatic cancer ,medicine ,Humans ,Breast ,MUC1 ,010405 organic chemistry ,Mucin ,Glycopeptides ,Antibodies, Monoclonal ,General Chemistry ,medicine.disease ,Molecular biology ,0104 chemical sciences ,Pancreatic Neoplasms ,Female - Abstract
In studies within the realm of cancer immunotherapy, the synthesis of exactly specified tumor-associated glycopeptide antigens is shown to be a key strategy for obtaining a highly selective biological reagent, that is, a monoclonal antibody that completely differentiates between tumor and normal epithelial cells and specifically marks the tumor cells in pancreas tumors. Mucin MUC1, which is overexpressed in many prevalent cancers, was identified as a promising target for this strategy. Tumor-associated MUC1 differs significantly from that expressed by normal cells, in particular by altered glycosylation. Structurally defined tumor-associated MUC1 cannot be isolated from tumor cells. We synthesized MUC1-glycopeptide vaccines and analyzed their structure-activity relationships in immunizations; a monoclonal antibody that specifically distinguishes between human normal and tumor epithelial cells was thus generated.
- Published
- 2015
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