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A Multimerized Form of Recombinant Human CD40 Ligand Supports Long-Term Activation and Proliferation of B Cells

A Multimerized Form of Recombinant Human CD40 Ligand Supports Long-Term Activation and Proliferation of B Cells

Maria A Garcia-Marquez, Alexander Shimabukuro-Vornhagen, Sebastian Theurich, Matthias Kochanek, Tanja Weber, Kerstin Wennhold, Alexandra Dauben, Andrzej Dzionek, Claudia Reinhard, Michael von Bergwelt-Baildon

Cytotherapy. 2014 Nov;16(11):1537-1544.

PMID: 25287602

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.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
IAR42413210	CD40L, Soluble human			Price

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