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2. Allergen-loaded strontium-doped hydroxyapatite spheres improve allergen-specific immunotherapy in mice

Author

Garbani, M., Xia, Wei, Rhyner, C., Prati, M., Scheynius, Annika, Malissen, B., Engqvist, Håkan, Maurer, M., Crameri, Reto, Terhorst-Molawi, Dorothea, Garbani, M., Xia, Wei, Rhyner, C., Prati, M., Scheynius, Annika, Malissen, B., Engqvist, Håkan, Maurer, M., Crameri, Reto, and Terhorst-Molawi, Dorothea

Abstract

Background Immunomodulatory interventions play a key role in the treatment of infections and cancer as well as allergic diseases. Adjuvants such as micro- and nanoparticles are often added to immunomodulatory therapies to enhance the triggered immune response. Here, we report the immunological assessment of novel and economically manufactured microparticle adjuvants, namely strontium-doped hydroxyapatite porous spheres (SHAS), which we suggest for the use as adjuvant and carrier in allergen-specific immunotherapy (ASIT). Methods and Results Scanning electron microscopy revealed that the synthesis procedure developed for the production of SHAS results in a highly homogeneous population of spheres. SHAS bound and released proteins such as ovalbumin (OVA) or the major cat allergen Fel d 1. SHAS-OVA were taken up by human monocyte-derived dendritic cells (mdDCs) and murine DCs and did not have any necrotic or apoptotic effects even at high densities. In a murine model of ASIT for allergic asthmatic inflammation we found that OVA released from subcutaneously injected SHAS-OVA led to a sustained stimulation of both CD4+ and CD8+ T-cells. ASIT with SHAS-OVA as compared to soluble OVA resulted in similar humoral responses but in a higher efficacy as assessed by symptom scoring. Conclusion We conclude that SHAS may constitute a suitable carrier and adjuvant for ASIT with great potential due to its unique protein-binding properties.

Published
2017
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3. Allergen-loaded strontium-doped hydroxyapatite spheres improve allergen-specific immunotherapy in mice

Author

Garbani, M, Xia, W, Rhyner, C, Prati, M, Scheynius, A, Malissen, B, Engqvist, H, Maurer, M, Crameri, R, Terhorst, D, Garbani, M, Xia, W, Rhyner, C, Prati, M, Scheynius, A, Malissen, B, Engqvist, H, Maurer, M, Crameri, R, and Terhorst, D

Abstract

Background Immunomodulatory interventions play a key role in the treatment of infections and cancer as well as allergic diseases. Adjuvants such as micro- and nanoparticles are often added to immunomodulatory therapies to enhance the triggered immune response. Here, we report the immunological assessment of novel and economically manufactured microparticle adjuvants, namely strontium-doped hydroxyapatite porous spheres (SHAS), which we suggest for the use as adjuvant and carrier in allergen-specific immunotherapy (ASIT). Methods and Results Scanning electron microscopy revealed that the synthesis procedure developed for the production of SHAS results in a highly homogeneous population of spheres. SHAS bound and released proteins such as ovalbumin (OVA) or the major cat allergen Fel d 1. SHAS-OVA were taken up by human monocyte-derived dendritic cells (mdDCs) and murine DCs and did not have any necrotic or apoptotic effects even at high densities. In a murine model of ASIT for allergic asthmatic inflammation we found that OVA released from subcutaneously injected SHAS-OVA led to a sustained stimulation of both CD4+ and CD8+ T-cells. ASIT with SHAS-OVA as compared to soluble OVA resulted in similar humoral responses but in a higher efficacy as assessed by symptom scoring. Conclusion We conclude that SHAS may constitute a suitable carrier and adjuvant for ASIT with great potential due to its unique protein-binding properties.

Published
2017

4. Allergen-loaded strontium-doped hydroxyapatite spheres improve allergen-specific immunotherapy in mice

Author

Garbani, M., Xia, W., Engkvist, H., Scheynius, A., Malissen, Bernard, Maurer, M., Crameri, R., Terhorst, D., Centre d'Immunologie de Marseille - Luminy (CIML), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU), Aix Marseille Université (AMU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), and VAFFIDES, Chantal

Subjects
[SDV.IMM] Life Sciences [q-bio]/Immunology, [SDV.IMM]Life Sciences [q-bio]/Immunology
Abstract

Annual Meeting of the European-Society-for-Dermatological-Research (ESDR), Munich, GERMANY, SEP 07-10, 2016; International audience; no abstract

Published
2016

7. A lectin-mediated resistance of higher fungi against predators and parasites

Author

Bleuler-Martinez, S, Butschi, A, Garbani, M, Wälti, M A, Wohlschlager, T, Potthoff, E, Sabotiĉ, J, Pohleven, J, Lüthy, P, Hengartner, M O, Aebi, M, Künzler, M, University of Zurich, and Künzler, M

Subjects
Acanthamoeba castellanii, Cytoplasm, Mycelium, Fungi, Feeding Behavior, 10124 Institute of Molecular Life Sciences, Fungal Proteins, 1105 Ecology, Evolution, Behavior and Systematics, 1311 Genetics, Aedes, Polysaccharides, Lectins, Escherichia coli, 570 Life sciences, biology, Animals, Fruiting Bodies, Fungal, Cloning, Molecular, Caenorhabditis elegans
Abstract

Fruiting body lectins are ubiquitous in higher fungi and characterized by being synthesized in the cytoplasm and up-regulated during sexual development. The function of these lectins is unclear. A lack of phenotype in sexual development upon inactivation of the respective genes argues against a function in this process. We tested a series of characterized fruiting body lectins from different fungi for toxicity towards the nematode Caenorhabditis elegans, the mosquito Aedes aegypti and the amoeba Acanthamoeba castellanii. Most of the fungal lectins were found to be toxic towards at least one of the three target organisms. By altering either the fungal lectin or the glycans of the target organisms, or by including soluble carbohydrate ligands as competitors, we demonstrate that the observed toxicity is dependent on the interaction between the fungal lectins and specific glycans in the target organisms. The toxicity was found to be dose-dependent such that low levels of lectin were no longer toxic but still led to food avoidance by C. elegans. Finally, we show, in an ecologically more relevant scenario, that challenging the vegetative mycelium of Coprinopsis cinerea with the fungal-feeding nematode Aphelenchus avenae induces the expression of the nematotoxic fruiting body lectins CGL1 and CGL2. Based on these findings, we propose that filamentous fungi possess an inducible resistance against predators and parasites mediated by lectins that are specific for glycans of these antagonists.

Published
2011
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8. Equine dendritic cells generated with horse serum have enhanced functionality in comparison to dendritic cells generated with fetal bovine serum.

Author

Ziegler A, Everett H, Hamza E, Garbani M, Gerber V, Marti E, and Steinbach F

Subjects
Animals, Cattle, Cell Differentiation genetics, Cell Survival drug effects, Cells, Cultured, Culture Media chemistry, Culture Media standards, Gene Expression Profiling veterinary, Gene Expression Regulation drug effects, Monocytes cytology, Cell Culture Techniques methods, Cell Differentiation drug effects, Culture Media pharmacology, Dendritic Cells cytology, Dendritic Cells drug effects, Horses, Serum metabolism
Abstract

Background: Dendritic cells are professional antigen-presenting cells that play an essential role in the initiation and modulation of T cell responses. They have been studied widely for their potential clinical applications, but for clinical use to be successful, alternatives to xenogeneic substances like fetal bovine serum (FBS) in cell culture need to be found. Protocols for the generation of dendritic cells ex vivo from monocytes are well established for several species, including horses. Currently, the gold standard protocol for generating dendritic cells from monocytes across various species relies upon a combination of GM-CSF and IL-4 added to cell culture medium which is supplemented with FBS. The aim of this study was to substitute FBS with heterologous horse serum. For this purpose, equine monocyte-derived dendritic cells (eqMoDC) were generated in the presence of horse serum or FBS and analysed for the effect on morphology, phenotype and immunological properties. Changes in the expression of phenotypic markers (CD14, CD86, CD206) were assessed during dendritic cell maturation by flow cytometry. To obtain a more complete picture of the eqMoDC differentiation and assess possible differences between FBS- and horse serum-driven cultures, a transcriptomic microarray analysis was performed. Lastly, immature eqMoDC were primed with a primary antigen (ovalbumin) or a recall antigen (tetanus toxoid) and, after maturation, were co-cultured with freshly isolated autologous CD5 + T lymphocytes to assess their T cell stimulatory capacity., Results: The microarray analysis demonstrated that eqMoDC generated with horse serum were indistinguishable from those generated with FBS. However, eqMoDC incubated with horse serum-supplemented medium exhibited a more characteristic dendritic cell morphology during differentiation from monocytes. A significant increase in cell viability was also observed in eqMoDC cultured with horse serum. Furthermore, eqMoDC generated in the presence of horse serum were found to be superior in their functional T lymphocyte priming capacity and to elicit significantly less non-specific proliferation., Conclusions: EqMoDC generated with horse serum-supplemented medium showed improved morphological characteristics, higher cell viability and exhibited a more robust performance in the functional T cell assays. Therefore, horse serum was found to be superior to FBS for generating equine monocyte-derived dendritic cells.

Published
2016
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