title: CD40L-Tri, a novel formulation of recombinant human CD40L that effectively activates B cells.
abstract: CD40L has a well-established role in enhancing the immunostimulatory capacity of normal and malignant B cells, but a formulation suitable for clinical use has not been widely available. Like other TNF family members, in vivo and in vitro activity of CD40L requires a homotrimeric configuration, and growing evidence suggests that bioactivity depends on higher-order clustering of CD40. We generated a novel formulation of human recombinant CD40L (CD40L-Tri) in which the CD40L extracellular domain and a trimerization motif are connected by a long flexible peptide linker. We demonstrate that CD40L-Tri significantly expands normal CD19+ B cells by over 20- to 30-fold over 14 days and induces B cells to become highly immunostimulatory antigen-presenting cells (APCs). Consistent with these results, CD40L-Tri-activated B cells could effectively stimulate antigen-specific T responses (against the influenza M1 peptide) from normal volunteers. In addition, CD40L-Tri could induce malignant B cells to become effective APCs, such that tumor-directed immune responses could be probed. Together, our studies demonstrate the potent immune-stimulatory effects of CD40L-Tri on B cells that enable their expansion of antigen-specific human T cells. The potent bioactivity of CD40L-Tri is related to its ability to self-multimerize, which may be facilitated by its long peptide linker.
authors.affiliation: Cancer Vaccine Center, Dana-Farber Cancer Institute, Boston, MA, USA.
authors.forename: Masayasu
authors.initials: M
authors.lastname: Naito
authors.forename: Ursula
authors.initials: U
authors.lastname: Hainz
authors.forename: Ute E
authors.initials: UE
authors.lastname: Burkhardt
authors.forename: Buyin
authors.initials: B
authors.lastname: Fu
authors.forename: Deborah
authors.initials: D
authors.lastname: Ahove
authors.forename: Kristen E
authors.initials: KE
authors.lastname: Stevenson
authors.forename: Mohini
authors.initials: M
authors.lastname: Rajasagi
authors.forename: Baogong
authors.initials: B
authors.lastname: Zhu
authors.forename: Anselmo
authors.initials: A
authors.lastname: Alonso
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Witten
authors.forename: Ken-Ichi
authors.initials: K
authors.lastname: Matsuoka
authors.forename: Donna
authors.initials: D
authors.lastname: Neuberg
authors.forename: Jonathan S
authors.initials: JS
authors.lastname: Duke-Cohan
authors.forename: Catherine J
authors.initials: CJ
authors.lastname: Wu
authors.forename: Gordon J
authors.initials: GJ
authors.lastname: Freeman
mesh_terms: D000328:Adult; D000938:Antigen-Presenting Cells / Q000187:drug effects / Q000276:immunology; D018941:Antigens, CD19 / Q000032:analysis; D000956:Antigens, Viral / Q000276:immunology; D001402:B-Lymphocytes / Q000187:drug effects* / Q000276:immunology; D023201:CD40 Ligand / Q000096:biosynthesis / Q000276:immunology / Q000494:pharmacology*; D002478:Cells, Cultured; D002626:Chemistry, Pharmaceutical; D006801:Humans; D015451:Leukemia, Lymphocytic, Chronic, B-Cell / Q000188:drug therapy / Q000276:immunology; D008213:Lymphocyte Activation / Q000187:drug effects / Q000276:immunology; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology / Q000494:pharmacology*; D013601:T-Lymphocytes / Q000187:drug effects / Q000276:immunology; D014763:Viral Matrix Proteins / Q000276:immunology
chemical_list: D018941:Antigens, CD19; D000956:Antigens, Viral; C488936:M1 protein, Influenza A virus; D011994:Recombinant Proteins; D014763:Viral Matrix Proteins; D023201:CD40 Ligand