Process development for the controlled expansion and differentiation of human primary B lymphocytes outside the human body. Part 2: Use in bioprocess technology

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The aim of the second project phase applied here, is to use the tools developed in the first project phase for the controlled ex vivo proliferation and differentiation of primary human B lymphocytes to prepare such cells for exemplary applications in bioprocess technology. Using natural abilities of the B lymphocytes such as: the ability to multiply quickly and then to pro-duce large amounts of protein (antibodies) (production type A: 'plasma cell'), the ability to per-sist for extended lengths of time in the body and remain biologically active (production type B: 'Living Drug'), as well as the ability to activate T lymphocytes in a targeted manner (produc-tion type C: 'antigen-presenting cell'), we intend to demonstrate the potential of such cells for applications in bioproduction and cell-based therapies. Each envisioned application/production types require different types of tailor-made B-cell subpopulations, sometimes in larger amounts. The challenge in realizing production type A is to decouple the proliferation of memory B cells and their differentiation into antibody-producing cells. The aim of the project in the sense of a 'demonstrator application' in case of production type A is the generation of an antiserum as a replacement for common blood-derived plasma preparations. In the case of production type B, the ability of B cells to persist in a metabolically active form in the hu-man body predestines such cells for use as 'living drugs', e.g. for protein replacement thera-py. The genetic modification of the B cells presents a great challenge in this case. Building on the results of the first project phase, non-viral transfection is to be used here, using the B cells subtype which is considered more suitable under process conditions. In the case of produc-tion type C, the aim is to provide tailor-made B-lymphocytes as antigen-presenting cells (APC). The activated T-lymphocytes thus represent the actual 'product' in this case. The main challenge in the case of production type C is the need to cultivate both types of lymphocytes together in a controlled manner. While production type A is primarily of interest for bioproduc-tion, production types B and C are also relevant for future medical applications. Within the scope of this project, however, only the basics of cell biology and process engineering are laid, right up to the first ‘demonstrator applications’.