There are no patents, products in development or marketed products to declare. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Ĭompeting interests: This study was funded in part by APEIRON Biologics AG. Hector Stiftung, Germany (grant number M57 recipient: HL) Deutsche Kinderkrebsstiftung, Germany (grant number: DKS 2014.05 A/B recipient: HL) and APEIRON Biologics AG, Austria (grant number: APN recipient: HL). This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.ĭata Availability: All relevant data are within the paper.įunding: This work was funded by University Medicine Greifswald, Germany (grant number 97237000 recipient: HL) H.W. Received: NovemAccepted: FebruPublished: March 11, 2016Ĭopyright: © 2016 Eger et al. PLoS ONE 11(3):Įditor: Pierre Busson, Gustave Roussy, FRANCE (2016) Generation and Characterization of a Human/Mouse Chimeric GD 2-Mimicking Anti-Idiotype Antibody Ganglidiximab for Active Immunotherapy against Neuroblastoma. This Ab may be useful to tailor immune responses to the paratope regions mimicking GD 2 overexpressed in NB.Ĭitation: Eger C, Siebert N, Seidel D, Zumpe M, Jüttner M, Brandt S, et al.
In summary, we report generation and characterization of a new human/mouse chimeric anti-Id Ab ganglidiximab for active immunotherapy against NB. Finally, ganglidiximab was successfully used as a protein vaccine in vivo to induce a GD 2-specific humoral immune response. Moreover, binding of anti-GD 2 Abs to the nominal antigen GD 2 as well as GD 2-specific Ab-mediated cytotoxicity (ADCC, CDC) was competitively inhibited by ganglidiximab. Ganglidiximab binding affinities to anti-GD 2 Abs were further determined by surface plasmon resonance technique. Binding of ganglidiximab to anti-GD 2 Abs of the 14.18 family as well as to NK-92tr cells expressing a GD 2-specific chimeric antigen receptor (scFv(ch14.18)-zeta) was shown using standard ELISA and flow cytometry analysis, respectively. After purification from supernatants, anti-idiotypic characteristics of ganglidiximab were demonstrated. 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. 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.
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.
We previously showed efficacy of ganglidiomab in vivo, a murine anti-idiotype (anti-Id) IgG1. Vaccination with proteins mimicking GD 2 that is highly expressed on neuroblastoma (NB) cells is a promising strategy in treatment of NB, a pediatric malignancy with poor prognosis.
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