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

作者：Garcia Marquez Maria A^*; Shimabukuro Vornhagen Alexander; Theurich Sebastian; Kochanek Matthias; Weber Tanja; Wennhold Kerstin; Dauben Alexandra; Dzionek Andrzej; Reinhard Claudia; von Bergwelt Baildon Michael

来源：Cytotherapy, 2014, 16(11): 1537-1544.

DOI：10.1016/j.jcyt.2014.05.011

• antigen presentation
• APC
• B cells
• CD4OL
• immunotherapy
• proliferation

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 carboxyfiuorescein 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.

• 出版日期2014-11