title: Generation and Characterization of a Human/Mouse Chimeric GD2-Mimicking Anti-Idiotype Antibody Ganglidiximab for Active Immunotherapy against Neuroblastoma.
abstract: Vaccination with proteins mimicking GD2 that is highly expressed on neuroblastoma (NB) cells is a promising strategy in treatment of NB, a pediatric malignancy with poor prognosis. We previously showed efficacy of ganglidiomab in vivo, a murine anti-idiotype (anti-Id) IgG1. In order to tailor immune responses to variable regions, we generated a new human/mouse chimeric anti-Id antibody (Ab) ganglidiximab by replacing murine constant fragments with corresponding human IgG1 regions. DNA sequences encoding for variable regions of heavy (VH) and light chains (VL) were synthesized by RT-PCR from total RNA of ganglidiomab-producing hybridoma cells and further ligated into mammalian expression plasmids with coding sequences for constant regions of human IgG1 heavy and light chains, respectively. We established a stable production cell line using Chinese hamster ovarian (CHO) cells co-transfected with two expression plasmids driving the expression of either ganglidiximab heavy or light chain. After purification from supernatants, anti-idiotypic characteristics of ganglidiximab were demonstrated. Binding of ganglidiximab to anti-GD2 Abs of the 14.18 family as well as to NK-92tr cells expressing a GD2-specific chimeric antigen receptor (scFv(ch14.18)-zeta) was shown using standard ELISA and flow cytometry analysis, respectively. Ganglidiximab binding affinities to anti-GD2 Abs were further determined by surface plasmon resonance technique. Moreover, binding of anti-GD2 Abs to the nominal antigen GD2 as well as GD2-specific Ab-mediated cytotoxicity (ADCC, CDC) was competitively inhibited by ganglidiximab. Finally, ganglidiximab was successfully used as a protein vaccine in vivo to induce a GD2-specific humoral immune response. In summary, we report generation and characterization of a new human/mouse chimeric anti-Id Ab ganglidiximab for active immunotherapy against NB. This Ab may be useful to tailor immune responses to the paratope regions mimicking GD2 overexpressed in NB.
authors.affiliation: Department of Pediatric Oncology and Hematology, University Medicine Greifswald, 17475, Greifswald, Germany.
authors.forename: Christin
authors.initials: C
authors.lastname: Eger
authors.affiliation: Department of Pediatric Oncology and Hematology, University Medicine Greifswald, 17475, Greifswald, Germany.
authors.forename: Nikolai
authors.initials: N
authors.lastname: Siebert
authors.affiliation: Department of Pediatric Oncology and Hematology, University Medicine Greifswald, 17475, Greifswald, Germany.
authors.forename: Diana
authors.initials: D
authors.lastname: Seidel
authors.affiliation: Department of Pediatric Oncology and Hematology, University Medicine Greifswald, 17475, Greifswald, Germany.
authors.forename: Maxi
authors.initials: M
authors.lastname: Zumpe
authors.affiliation: Department of Pediatric Oncology and Hematology, University Medicine Greifswald, 17475, Greifswald, Germany.
authors.forename: Madlen
authors.initials: M
authors.lastname: Jüttner
authors.affiliation: ZIK HIKE, Center for Innovation Competence: Humoral Immune Reactions in Cardiovascular Diseases, University of Greifswald, Greifswald, Germany.
authors.forename: Sven
authors.initials: S
authors.lastname: Brandt
authors.affiliation: ZIK HIKE, Center for Innovation Competence: Humoral Immune Reactions in Cardiovascular Diseases, University of Greifswald, Greifswald, Germany.
authors.forename: Hans-Peter
authors.initials: HP
authors.lastname: Müller
authors.affiliation: Department of Pediatric Oncology and Hematology, University Medicine Greifswald, 17475, Greifswald, Germany.
authors.forename: Holger N
authors.initials: HN
authors.lastname: Lode
mesh_terms: D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000235:genetics / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000008:administration & dosage* / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D000915:Antibody Affinity; D016466:CHO Cells; D019496:Cancer Vaccines / Q000008:administration & dosage* / Q000235:genetics / Q000378:metabolism*; D006224:Cricetinae; D003412:Cricetulus; D003602:Cytotoxicity, Immunologic; D006801:Humans; D016233:Immunotherapy, Active; D007694:Killer Cells, Natural; D051379:Mice; D009447:Neuroblastoma / Q000276:immunology / Q000628:therapy
chemical_list: D000888:Antibodies, Anti-Idiotypic; D000911:Antibodies, Monoclonal; D019496:Cancer Vaccines; C582619:ganglidiomab; C112746:dinutuximab