title: A multimerized form of recombinant human CD40 ligand supports long-term activation and proliferation of B cells.
abstract: OBJECTIVE
  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.
authors.affiliation: Cologne Interventional Immunology, University Hospital of Cologne, Cologne, Germany. Electronic address: maria.garcia-marquez@uk-koeln.de.
authors.forename: Maria A
authors.initials: MA
authors.lastname: Garcia-Marquez
authors.affiliation: Cologne Interventional Immunology, University Hospital of Cologne, Cologne, Germany; Stem Cell Transplantation Program, Department I of Internal Medicine, University Hospital of Cologne, Cologne, Germany.
authors.forename: Alexander
authors.initials: A
authors.lastname: Shimabukuro-Vornhagen
authors.affiliation: Cologne Interventional Immunology, University Hospital of Cologne, Cologne, Germany; Stem Cell Transplantation Program, Department I of Internal Medicine, University Hospital of Cologne, Cologne, Germany; Department of Mouse Genetics and Metabolism, Institute for Genetics, University of Cologne, Cologne, Germany; Max-Planck-Institute for Neurological Research, Cologne, Germany.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Theurich
authors.affiliation: Cologne Interventional Immunology, University Hospital of Cologne, Cologne, Germany; Stem Cell Transplantation Program, Department I of Internal Medicine, University Hospital of Cologne, Cologne, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Kochanek
authors.affiliation: Cologne Interventional Immunology, University Hospital of Cologne, Cologne, Germany.
authors.forename: Tanja
authors.initials: T
authors.lastname: Weber
authors.affiliation: Cologne Interventional Immunology, University Hospital of Cologne, Cologne, Germany.
authors.forename: Kerstin
authors.initials: K
authors.lastname: Wennhold
authors.affiliation: Miltenyi Biotec GmbH, Bergisch Gladbach, Germany.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Dauben
authors.affiliation: Miltenyi Biotec GmbH, Bergisch Gladbach, Germany.
authors.forename: Andrzej
authors.initials: A
authors.lastname: Dzionek
authors.affiliation: Miltenyi Biotec GmbH, Bergisch Gladbach, Germany.
authors.forename: Claudia
authors.initials: C
authors.lastname: Reinhard
authors.affiliation: Cologne Interventional Immunology, University Hospital of Cologne, Cologne, Germany; Stem Cell Transplantation Program, Department I of Internal Medicine, University Hospital of Cologne, Cologne, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: von Bergwelt-Baildon
mesh_terms: D000818:Animals; D000938:Antigen-Presenting Cells / Q000276:immunology; D001402:B-Lymphocytes / Q000166:cytology / Q000276:immunology*; D023201:CD40 Ligand / Q000276:immunology* / Q000378:metabolism; D049109:Cell Proliferation; D006801:Humans; D007167:Immunotherapy / Q000379:methods; D008213:Lymphocyte Activation / Q000276:immunology; D051379:Mice; D041681:NIH 3T3 Cells; D013601:T-Lymphocytes / Q000276:immunology; D014162:Transfection
chemical_list: D023201:CD40 Ligand
keywords: APC; B cells; CD40L; antigen presentation; immunotherapy; proliferation