TY - JOUR A1 - Groeber, Florian A1 - Engelhardt, Lisa A1 - Lange, Julia A1 - Kurdyn, Szymon A1 - Schmid, Freia F. A1 - Rücker, Christoph A1 - Mielke, Stephan A1 - Walles, Heike A1 - Hansmann, Jan T1 - A First Vascularized Skin Equivalent as an Alternative to Animal Experimentation JF - ALTEX - Alternatives to Animal Experimentation N2 - Tissue-engineered skin equivalents mimic key aspects of the human skin, and can thus be employed as wound coverage for large skin defects or as in vitro test systems as an alternative to animal models. However, current skin equivalents lack a functional vasculature limiting clinical and research applications. This study demonstrates the generation of a vascularized skin equivalent with a perfused vascular network by combining a biological vascularized scaffold (BioVaSc) based on a decellularized segment of a porcine jejunum and a tailored bioreactor system. Briefly, the BioVaSc was seeded with human fibroblasts, keratinocytes, and human microvascular endothelial cells. After 14 days at the air-liquid interface, hematoxylin & eosin and immunohistological staining revealed a specific histological architecture representative of the human dermis and epidermis including a papillary-like architecture at the dermal-epidermal-junction. The formation of the skin barrier was measured non-destructively using impedance spectroscopy. Additionally, endothelial cells lined the walls of the formed vessels that could be perfused with a physiological volume flow. Due to the presence of a complex in-vivo-like vasculature, the here shown skin equivalent has the potential for skin grafting and represents a sophisticated in vitro model for dermatological research. KW - alternative to animal testing KW - skin equivalents KW - tissue engineering KW - vascularization Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-164438 VL - 33 IS - 4 ER - TY - JOUR A1 - Roy, Denis Claude A1 - Lachance, Sylvie A1 - Cohen, Sandra A1 - Delisle, Jean-Sébastien A1 - Kiss, Thomas A1 - Sauvageau, Guy A1 - Busque, Lambert A1 - Ahmad, Imran A1 - Bernard, Lea A1 - Bambace, Nadia A1 - Boumédine, Radia S. A1 - Guertin, Marie-Claude A1 - Rezvani, Katayoun A1 - Mielke, Stephan A1 - Perreault, Claude A1 - Roy, Jean T1 - Allodepleted T-cell immunotherapy after haploidentical haematopoietic stem cell transplantation without severe acute graft-versus-host disease (GVHD) in the absence of GVHD prophylaxis JF - British Journal of Haematology N2 - Graft-versus-host disease (GVHD) is a major cause of transplant-related mortality (TRM) after allogeneic haematopoietic stem cell transplantation (HSCT) and presents a challenge in haploidentical HSCT. GVHD may be prevented by ex vivo graft T-cell depletion or in vivo depletion of proliferating lymphocytes. However, both approaches pose significant risks, particularly infections and relapse, compromising survival. A photodepletion strategy to eliminate alloreactive T cells from mismatched donor lymphocyte infusions (enabling administration without immunosuppression), was used to develop ATIR101, an adjunctive therapy for use after haploidentical HSCT. In this phase I dose-finding study, 19 adults (median age: 54 years) with high-risk haematological malignancies were treated with T-cell-depleted human leucocyte antigen-haploidentical myeloablative HSCT followed by ATIR101 at doses of 1 x 10(4)-5 x 10(6) CD3(+) cells/kg (median 31 days post-transplant). No patient received post-transplant immunosuppression or developed grade III/IV acute GVHD, demonstrating the feasibility of ATIR101 infusion for evaluation in two subsequent phase 2 studies. Additionally, we report long-term follow -up of patients treated with ATIR101 in this study. At 1 year, all 9 patients receiving doses of 0 center dot 3-2 x 10(6) CD3(+) cells/kg ATIR101 remained free of serious infections and after more than 8 years, TRM was 0%, relapse-related mortality was 33% and overall survival was 67% in these patients. KW - haematopoietic stem cell KW - stem cell transplantation KW - graft-versus-host-disease KW - cell therapy and immunotherapy Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-227075 VL - 186 IS - 5 ER - TY - JOUR A1 - Wagner-Drouet, Eva A1 - Teschner, Daniel A1 - Wolschke, Christine A1 - Schäfer-Eckart, Kerstin A1 - Gärtner, Johannes A1 - Mielke, Stephan A1 - Schreder, Martin A1 - Kobbe, Guido A1 - Hilgendorf, Inken A1 - Klein, Stefan A1 - Verbeek, Mareike A1 - Ditschkowski, Markus A1 - Koch, Martina A1 - Lindemann, Monika A1 - Schmidt, Traudel A1 - Rascle, Anne A1 - Barabas, Sascha A1 - Deml, Ludwig A1 - Wagner, Ralf A1 - Wolff, Daniel T1 - Comparison of cytomegalovirus-specific immune cell response to proteins versus peptides using an IFN-γ ELISpot assay after hematopoietic stem cell transplantation JF - Diagnostics N2 - Cytomegalovirus (CMV) infection is a major cause of morbidity and mortality following hematopoietic stem cell transplantation (HSCT). Measuring CMV-specific cellular immunity may improve the risk stratification and management of patients. IFN-γ ELISpot assays, based on the stimulation of peripheral blood mononuclear cells with CMV pp65 and IE-1 proteins or peptides, have been validated in clinical settings. However, it remains unclear to which extend the T-cell response to synthetic peptides reflect that mediated by full-length proteins processed by antigen-presenting cells. We compared the stimulating ability of pp65 and IE-1 proteins and corresponding overlapping peptides in 16 HSCT recipients using a standardized IFN-γ ELISpot assay. Paired qualitative test results showed an overall 74.4% concordance. Discordant results were mainly due to low-response tests, with one exception. One patient with early CMV reactivation and graft-versus-host disease, sustained CMV DNAemia and high CD8\(^+\) counts showed successive negative protein-based ELISpot results but a high and sustained response to IE-1 peptides. Our results suggest that the response to exogenous proteins, which involves their uptake and processing by antigen-presenting cells, more closely reflects the physiological response to CMV infection, while the response to exogenous peptides may lead to artificial in vitro T-cell responses, especially in strongly immunosuppressed patients. KW - CMV KW - CMV-specific cellular immunity KW - hematopoietic stem cell transplantation KW - recall antigen KW - peptide KW - immune monitoring KW - IFN-γ ELISpot KW - T cells KW - antigen processing and presentation KW - immunosuppression Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-228843 SN - 2075-4418 VL - 11 IS - 2 ER - TY - JOUR A1 - Luber, Verena A1 - Lutz, Mathias A1 - Thiede, Christian A1 - Haferlach, Claudia A1 - Dürk, Heinz Albert A1 - Einsele, Hermann A1 - Grigoleit, Götz Ulrich A1 - Mielke, Stephan T1 - Donor-cell leukemia with novel genetic features 2 years after sex-mismatched T cell-depleted haploidentical stem cell transplantation JF - Annals of Hematology N2 - No abstract available. KW - donor-cell leukemia Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-232596 SN - 0939-5555 VL - 99 ER - TY - JOUR A1 - Luber, Verena A1 - Lutz, Mathias A1 - Abele-Horn, Marianne A1 - Einsele, Hermann A1 - Grigoleit, Götz Ulrich A1 - Mielke, Stephan T1 - Excretion of Ascaris lumbricoides following reduced‐intensity allogeneic hematopoietic stem cell transplantation and consecutive treatment with mebendazole JF - Transplant Infectious Disease N2 - Here, we present the unique case of a 51‐year‐old German patient with multiple myeloma excreting Ascaris lumbricoides in his stool five weeks after allogeneic hematopoietic stem cell transplantation. Stool analysis remained negative for the presence of eggs, and there was no eosinophilia in the peripheral blood at any time around stem cell transplantation. The patient was commenced on a three‐day treatment with mebendazole, which was well tolerated. No serious interactions with the concomitant post‐transplant medication or negative effects on the hematopoiesis were observed, and the myeloma still is in complete remission. To our knowledge, this is the first report on excretion of A lumbricoides in the context of allogeneic stem cell transplantation. The case is remarkable with view to the fact that the parasite has supposedly survived all courses of myeloma treatment including autologous and allogeneic conditioning. Parasitosis with A lumbricoides has a worldwide prevalence of about a billion and is extremely rare in northern Europe. Possibly the patient got infected during a trip to Egypt years before multiple myeloma was diagnosed. KW - sirolimus KW - mycophenolic acid KW - multiple myeloma KW - mebendazole KW - hematopoietic stem cell transplantation KW - Ascaris lumbricoides Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-219608 SN - 1399-3062 VL - 22 IS - 1 ER - TY - JOUR A1 - Wallstabe, Julia A1 - Bussemer, Lydia A1 - Groeber-Becker, Florian A1 - Freund, Lukas A1 - Alb, Mirian A1 - Dragan, Mariola A1 - Waaga-Gasser, Ana Maria A1 - Jakubietz, Rafael A1 - Kneitz, Hermann A1 - Rosenwald, Andreas A1 - Rebhan, Silke A1 - Walles, Heike A1 - Mielke, Stephan T1 - Inflammation-Induced Tissue Damage Mimicking GvHD in Human Skin Models as Test Platform for Immunotherapeutics JF - ALTEX N2 - Due to the rapidly increasing development and use of cellular products, there is a rising demand for non-animal-based test platforms to predict, study and treat undesired immunity. Here, we generated human organotypic skin models from human biopsies by isolating and expanding keratinocytes, fibroblasts and microvascular endothelial cells and seeding these components on a collagen matrix or a biological vascularized scaffold matrix in a bioreactor. We then were able to induce inflammation-mediated tissue damage by adding pre-stimulated, mismatched allogeneic lymphocytes and/or inflammatory cytokine-containing supernatants histomorphologically mimicking severe graft versus host disease (GvHD) of the skin. This could be prevented by the addition of immunosuppressants to the models. Consequently, these models harbor a promising potential to serve as a test platform for the prediction, prevention and treatment of GvHD. They also allow functional studies of immune effectors and suppressors including but not limited to allodepleted lymphocytes, gamma-delta T cells, regulatory T cells and mesenchymal stromal cells, which would otherwise be limited to animal models. Thus, the current test platform, developed with the limitation that no professional antigen presenting cells are in place, could greatly reduce animal testing for investigation of novel immune therapies. KW - inflammation-induced tissue demage KW - immunotherapeutics Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-229974 VL - 37 IS - 3 ER - TY - JOUR A1 - Stringaris, Kate A1 - Sekine, Takuya A1 - Khoder, Ahmad A1 - Alsuliman, Abdullah A1 - Razzaghi, Bonni A1 - Sargeant, Ruhena A1 - Pavlu, Jiri A1 - Brisley, Gill A1 - de Lavallade, Hugues A1 - Sarvaria, Anushruthi A1 - Sarvaria, Anushruthi A1 - Mielke, Stephan A1 - Apperley, Jane F. A1 - Shpall, Elisabeth J. A1 - Barrett, A. John A1 - Rezvani, Katayoun T1 - Leukemia-induced phenotypic and functional defects in natural killer cells predict failure to achieve remission in acute myeloid leukemia JF - Haematologica N2 - The majority of patients with acute myeloid leukemia will relapse, and older patients often fail to achieve remission with induction chemotherapy. We explored the possibility that leukemic suppression of innate immunity might contribute to treatment failure. Natural killer cell phenotype and function was measured in 32 consecutive acute myeloid leukemia patients at presentation, including 12 achieving complete remission. Compared to 15 healthy age-matched controls, natural killer cells from acute myeloid leukemia patients were abnormal at presentation, with downregulation of the activating receptor NKp46 (P=0.007) and upregulation of the inhibitory receptor NKG2A (P=0.04). Natural killer cells from acute myeloid leukemia patients had impaired effector function against autologous blasts and K562 targets, with significantly reduced CD107a degranulation, TNF-alpha and IFN-gamma production. Failure to achieve remission was associated with NKG2A overexpression and reduced TNF-alpha production. These phenotypic and functional abnormalities were partially restored in the 12 patients achieving remission. In vitro co-incubation of acute myeloid leukemia blasts with natural killer cells from healthy donors induced significant impairment in natural killer cell TNF-alpha and IFN-gamma production (P=0.02 and P=0.01, respectively) against K562 targets and a trend to reduced CD107a degranulation (P=0.07). Under transwell conditions, the inhibitory effect of AML blasts on NK cytotoxicity and effector function was still present, and this inhibitory effect was primarily mediated by IL-10. These results suggest that acute myeloid leukemia blasts induce long-lasting changes in natural killer cells, impairing their effector function and reducing the competence of the innate immune system, favoring leukemia survival. KW - resting NK cells KW - acute myelogenous leukemia KW - virus-infected cells KW - cytotoxicity receptors KW - inhibitory receptors KW - ligand incompatibility KW - activating receptors KW - MHC molecules KW - missing self KW - class-I Y1 - 2014 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-116550 SN - 1592-8721 VL - 99 IS - 5 ER -