Studies on Idiotype-driven T-B collaboration
Aim: to study the biological processes involved in T cell-B cell collaboration which is an important part of adaptive immunity, especially of antibody production. The change that is applied now will facilitate imaging in situ (IVIS imager) and post-mortem (FACS and IHC) of transferred cells irrespective of their differentiation status, in cases where the specific cell surface receptor is down-regulated (i.e. not detecable).
Expected distress: these experiments are not aimed at modelling disease, rather to study principles of basic biology. We do not expect pathology to develop during the course of the experiments. Distress will be related to routine experimental procedures, such as anasethesia, i.v. or i.p. injections, and saphenous vein blood sampling. The expression of fluorescent proteins are not expected to induce any distress or increase risk for developing pathology.Such fluorescent protein are non-toxic and non-immunogenic and are derived from living organisms. Their use is widespread in molecular biology.
Expected Scientific and societal benefit: to increase our understanding relating to the biological principles of T-B collaboration, B cell maturation, memory formation and antibody production. Furthermore, as some of these processes contribute to the development of autoimmune diseases, the data from the experiments will increase our understanding of aspects of autoimmune disease. Furthermore, the role of regulatory T cells in tolerance will be investigated. The use of the fluorescent strains will enable better imaging modalities and allow us to extract more relevant biological information from each mouse.
Animals: we will use transgenic, knock-in and gene modified mice. All are maintained at our facility. We aim to use a maximum of 2346 animals in four years.
The 3 R`s: we will use in vitro experiments to complement our studies, where possible. We will reduce the number of experimental animals to a minimum. Improvement: we will rely largely on the experience we obtained during the course of previous experiments. These were developed under FOTSID 7414, but also other protocols. We know how many B cells, T cells and BCR ligand are needed for a productive response, for example.
Expected distress: these experiments are not aimed at modelling disease, rather to study principles of basic biology. We do not expect pathology to develop during the course of the experiments. Distress will be related to routine experimental procedures, such as anasethesia, i.v. or i.p. injections, and saphenous vein blood sampling. The expression of fluorescent proteins are not expected to induce any distress or increase risk for developing pathology.Such fluorescent protein are non-toxic and non-immunogenic and are derived from living organisms. Their use is widespread in molecular biology.
Expected Scientific and societal benefit: to increase our understanding relating to the biological principles of T-B collaboration, B cell maturation, memory formation and antibody production. Furthermore, as some of these processes contribute to the development of autoimmune diseases, the data from the experiments will increase our understanding of aspects of autoimmune disease. Furthermore, the role of regulatory T cells in tolerance will be investigated. The use of the fluorescent strains will enable better imaging modalities and allow us to extract more relevant biological information from each mouse.
Animals: we will use transgenic, knock-in and gene modified mice. All are maintained at our facility. We aim to use a maximum of 2346 animals in four years.
The 3 R`s: we will use in vitro experiments to complement our studies, where possible. We will reduce the number of experimental animals to a minimum. Improvement: we will rely largely on the experience we obtained during the course of previous experiments. These were developed under FOTSID 7414, but also other protocols. We know how many B cells, T cells and BCR ligand are needed for a productive response, for example.