title: Vaccination with anti-idiotype antibody ganglidiomab mediates a GD(2)-specific anti-neuroblastoma immune response.
abstract: OBJECTIVE
  Immunotherapy targeting disialoganglioside GD(2) emerges as an important treatment option for neuroblastoma, a pediatric malignancy characterized by poor outcome. Here, we report the induction of a GD(2)-specific immune response with ganglidiomab, a new anti-idiotype antibody to anti-GD(2) antibodies of the 14.18 family.
  
  
  RESULTS
  Ganglidiomab was generated following immunization of Balb/c mice with 14G2a, and splenocytes were harvested to generate hybridoma cells. Clones were screened by ELISA for mouse antibody binding to hu14.18. One positive clone was selected to purify and characterize the secreted IgG protein (κ, IgG(1)). This antibody bound to anti-GD(2) antibodies 14G2a, ch14.18/CHO, hu14.18, and to immunocytokines ch14.18-IL2 and hu14.18-IL2 as well as to NK-92 cells expressing scFv(ch14.18)-zeta receptor. Binding of these anti-GD(2) antibodies to the nominal antigen GD(2) as well as GD(2)-specific lysis of neuroblastoma cells by NK-92-scFv(ch14.18)-zeta cells was competitively inhibited by ganglidiomab, proving GD(2) surrogate function and anti-idiotype characteristics. The dissociation constants of ganglidiomab from anti-GD(2) antibodies ranged from 10.8 ± 5.01 to 53.5 ± 1.92 nM as determined by Biacore analyses. The sequences of framework and complementarity-determining regions of ganglidiomab were identified. Finally, we demonstrated induction of a GD(2)-specific humoral immune response after vaccination of mice with ganglidiomab effective in mediating GD(2)-specific killing of neuroblastoma cells.
  
  
  CONCLUSIONS
  We generated and characterized a novel anti-idiotype antibody ganglidiomab and demonstrated activity against neuroblastoma.
authors.affiliation: Pediatric Hematology and Oncology, University Medicine Greifswald, Ferdinand-Sauerbruchstrasse 1, 17475 Greifswald, Germany. lode@uni-greifswald.de
authors.forename: Holger N
authors.initials: HN
authors.lastname: Lode
authors.forename: Manuela
authors.initials: M
authors.lastname: Schmidt
authors.forename: Diana
authors.initials: D
authors.lastname: Seidel
authors.forename: Nicole
authors.initials: N
authors.lastname: Huebener
authors.forename: Diana
authors.initials: D
authors.lastname: Brackrock
authors.forename: Matthias
authors.initials: M
authors.lastname: Bleeke
authors.forename: Daniel
authors.initials: D
authors.lastname: Reker
authors.forename: Sven
authors.initials: S
authors.lastname: Brandt
authors.forename: Hans-Peter
authors.initials: HP
authors.lastname: Mueller
authors.forename: Christiane
authors.initials: C
authors.lastname: Helm
authors.forename: Nikolai
authors.initials: N
authors.lastname: Siebert
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000008:administration & dosage / Q000737:chemistry / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000008:administration & dosage / Q000737:chemistry / Q000276:immunology*; D001483:Base Sequence; D001667:Binding, Competitive / Q000276:immunology; D019496:Cancer Vaccines / Q000276:immunology*; D045744:Cell Line, Tumor; D005732:Gangliosides / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D051379:Mice; D008969:Molecular Sequence Data; D009447:Neuroblastoma / Q000276:immunology* / Q000628:therapy; D011485:Protein Binding / Q000276:immunology; D016415:Sequence Alignment
chemical_list: D000888:Antibodies, Anti-Idiotypic; D000911:Antibodies, Monoclonal; D019496:Cancer Vaccines; D005732:Gangliosides; C582619:ganglidiomab; C019403:ganglioside, GD2