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A multimerized form of recombinant human CD40 ligand supports long-term activation and proliferation of B cells

MPS-Authors

Garcia-Marquez,  M. A.
Max Planck Institute for Biology of Ageing, Max Planck Society;

Shimabukuro-Vornhagen,  A.
Max Planck Institute for Biology of Ageing, Max Planck Society;

Theurich,  S.
Max Planck Institute for Biology of Ageing, Max Planck Society;

Kochanek,  M.
Max Planck Institute for Biology of Ageing, Max Planck Society;

Weber,  T.
Max Planck Institute for Biology of Ageing, Max Planck Society;

Wennhold,  K.
Max Planck Institute for Biology of Ageing, Max Planck Society;

Dauben,  A.
Max Planck Institute for Biology of Ageing, Max Planck Society;

Dzionek,  A.
Max Planck Institute for Biology of Ageing, Max Planck Society;

Reinhard,  C.
Max Planck Institute for Biology of Ageing, Max Planck Society;

von Bergwelt-Baildon,  M.
Max Planck Institute for Biology of Ageing, Max Planck Society;

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Citation

Garcia-Marquez, M. A., Shimabukuro-Vornhagen, A., Theurich, S., Kochanek, M., Weber, T., Wennhold, K., et al. (2014). A multimerized form of recombinant human CD40 ligand supports long-term activation and proliferation of B cells. Cytotherapy, 16(11), 1537-44. doi:10.1016/j.jcyt.2014.05.011.


Cite as: http://hdl.handle.net/11858/00-001M-0000-0028-58E2-F
Abstract
BACKGROUND AIMS: CD40-activated B cells have long been studied as potent antigen-presenting cells that can potentially be used for cancer immunotherapy. Nevertheless, their use in human clinical trials has been limited by the lack of a Good Manufacturing Practice-grade soluble human CD40 ligand that is able to induce activation and proliferation of primary B cells. We describe an in vitro method to effectively generate and expand B cells through the use of a multimerized form of human recombinant CD40 ligand (rCD40L). METHODS: Human B cells were isolated from healthy donors and cultivated with either rCD40L or on a monolayer of murine NIH3T3 cells stably expressing human CD40L (NIH3T3/tCD40L) as a widely used standard method. Morphology, expansion rate, immune phenotype and antigen presentation function were assessed. RESULTS: B cells efficiently proliferated in response to rCD40L over 14 days of culture in comparable amounts to NIH3T3/tCD40L. B-cell division in response to CD40L was also confirmed by carboxyfluorescein succinimidyl ester dilution. Moreover, rCD40L induced on B cells upregulation of co-stimulatory molecules essential for antigen presentation. Additionally, proliferation of T cells from allogeneic healthy volunteers confirmed the immunostimulatory capacities of CD40-activated B cells. CONCLUSIONS: We demonstrated that B cells with potent antigen presentation capacity can be generated and expanded by use of a non-xenogeneic form of CD40L that could be implemented in future human clinical settings.