title: Targeted delivery of CD40L promotes restricted activation of antigen-presenting cells and induction of cancer cell death.
abstract: BACKGROUND
  Stimulation of CD40 can augment anti-cancer T cell immune responses by triggering effective activation and maturation of antigen-presenting cells (APCs). Although CD40 agonists have clinical activity in humans, the associated systemic activation of the immune system triggers dose-limiting side-effects.
  
  
  METHODS
  To increase the tumor selectivity of CD40 agonist-based therapies, we developed an approach in which soluble trimeric CD40L (sCD40L) is genetically fused to tumor targeting antibody fragments, yielding scFv:CD40L fusion proteins. We hypothesized that scFv:CD40L fusion proteins would have reduced CD40 agonist activity similar to sCD40L but will be converted to a highly agonistic membrane CD40L-like form of CD40L upon anchoring to cell surface exposed antigen via the scFv domain.
  
  
  RESULTS
  Targeted delivery of CD40L to the carcinoma marker EpCAM on carcinoma cells induced dose-dependent paracrine maturation of DCs ~20-fold more effective than a non-targeted control scFv:CD40L fusion protein. Similarly, targeted delivery of CD40L to the B cell leukemia marker CD20 induced effective paracrine maturation of DCs. Of note, the CD20-selective delivery of CD40L also triggered loss of cell viability in certain B cell leukemic cell lines as a result of CD20-induced apoptosis.
  
  
  CONCLUSIONS
  Targeted delivery of CD40L to cancer cells is a promising strategy that may help to trigger cancer-localized activation of CD40 and can be modified to exert additional anti-cancer activity via the targeting domain.
authors.forename: Kim L
authors.initials: KL
authors.lastname: Brunekreeft
authors.forename: Corinna
authors.initials: C
authors.lastname: Strohm
authors.forename: Marloes J
authors.initials: MJ
authors.lastname: Gooden
authors.forename: Anna A
authors.initials: AA
authors.lastname: Rybczynska
authors.forename: Hans W
authors.initials: HW
authors.lastname: Nijman
authors.forename: Götz U
authors.initials: GU
authors.lastname: Grigoleit
authors.forename: Wijnand
authors.initials: W
authors.lastname: Helfrich
authors.forename: Edwin
authors.initials: E
authors.lastname: Bremer
authors.forename: Daniela
authors.initials: D
authors.lastname: Siegmund
authors.forename: Harald
authors.initials: H
authors.lastname: Wajant
authors.affiliation: Department of Obstetrics and Gynecology, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands. m.de.bruyn@umcg.nl.
authors.forename: Marco
authors.initials: M
authors.lastname: de Bruyn
mesh_terms: D058846:Antibodies, Monoclonal, Murine-Derived / Q000494:pharmacology; D018951:Antigens, CD20 / Q000235:genetics / Q000378:metabolism; D000951:Antigens, Neoplasm / Q000235:genetics / Q000378:metabolism; D000970:Antineoplastic Agents / Q000494:pharmacology; D001402:B-Lymphocytes / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D023201:CD40 Ligand / Q000235:genetics* / Q000378:metabolism; D015815:Cell Adhesion Molecules / Q000235:genetics / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D003713:Dendritic Cells / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D000071858:Epithelial Cell Adhesion Molecule; D015870:Gene Expression; D057809:HEK293 Cells; D006801:Humans; D058990:Molecular Targeted Therapy; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D000069283:Rituximab; D057127:Single-Chain Antibodies / Q000235:genetics* / Q000378:metabolism
chemical_list: D058846:Antibodies, Monoclonal, Murine-Derived; D018951:Antigens, CD20; D000951:Antigens, Neoplasm; D000970:Antineoplastic Agents; D015815:Cell Adhesion Molecules; C499379:EPCAM protein, human; D000071858:Epithelial Cell Adhesion Molecule; D011993:Recombinant Fusion Proteins; D057127:Single-Chain Antibodies; D023201:CD40 Ligand; D000069283:Rituximab
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