TY - JOUR A1 - Wand, Inga A1 - Holzlöhner, Pamela A1 - Neupert, Steffi A1 - Micheel, Burkhard A1 - Heilmann, Katja T1 - Cooperation of dendritic cells with naive lymphocyte populations to induce the generation of antigen-specific antibodies in vitro JF - Journal of biotechnology N2 - The production of monoclonal antibodies by hybridoma technology is dependent on lymphocytes taken from vertebrates which have to be immunized against the corresponding antigen. We present here our first experiments which should allow the replacement of this in vivo immunization step by an in vitro immunization procedure. This work provides new possibilities for the specific activation of immune cells in order to use them for the generation of antibodies which are not of murine origin. Bone marrow-derived dendritic cells were loaded with antigen and co-cultured with naive T and B lymphocytes of non-immunized mice. The interaction and activation of the different cell types were investigated by measuring the expression of specific cell surface markers, the release of activation-dependent interleukins and the secretion of antigen-specific antibodies. We could demonstrate that dendritic cells process and present antigen fragments and activate T cells, that T cells proliferate and release activation-induced interleukins, and that B cells maturate under the influence of activated T cells and secrete antigen-specific antibodies. KW - In vitro immunization KW - Activation of dendritic cells KW - Interaction of T and B cells with antigen-presenting cells KW - Induction of antibody responses Y1 - 2011 U6 - https://doi.org/10.1016/j.jbiotec.2011.09.002 SN - 0168-1656 VL - 156 IS - 3 SP - 173 EP - 181 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Michelchen, Sophia A1 - Micheel, Burkhard A1 - Hanack, Katja T1 - In vitro immunization approach to generate specific murine monoclonal IgG antibodies JF - Journal of immunological methods : JIM N2 - Generating a monoclonal antibody to date is a time intense process that requires immunization of laboratory animals. The transfer of the humoral immune response into in vitro settings enables a shortening of this process and circumvents the necessity of in vivo immunization. However, to orchestrate the complex interplay of dendritic cells, T and B lymphocytes in vitro is very challenging. We therefore aimed for a simplified approach focusing on the protagonist of antibody production: the B lymphocyte. We activated purified murine B lymphocytes alone in vitro by using combinations of antigen and stimuli. We were able to induce a specific antibody response within ten days of culture against a viral coat protein as model antigen. Antibodies were of both IgM and IgG subclass. The stimulated B lymphocytes were transformed into permanently antibody-producing hybridomas by cell fusion technology. We furthermore used this method to induce a specific antibody response against L. pneumophila in vitro. We thus established a useful and effective in vitro protocol to generate monoclonal antibodies. By overcoming the necessity of in vivo immunization this protocol may be the first step towards a universal strategy to generate antibodies from various species. KW - Monoclonal antibody KW - Hybridoma technology KW - In vitro immunization KW - B cell activation Y1 - 2021 U6 - https://doi.org/10.1016/j.jim.2021.113149 SN - 0022-1759 SN - 1872-7905 VL - 499 PB - Elsevier CY - Amsterdam ER -