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.

Structural Mimicry of the Dengue Virus Envelope Glycoprotein Revealed by the Crystallographic Study of an Idiotype–Anti-idiotype Fab Complex

“…3 in reference 3), and several of these antibodies were not derived from the same D1.3 clone. More recently, efforts to develop "idiotypic vaccines" for cancer treatment have been reported (15,16). Racotumomab, an antiidiotypic antibody targeting the NeuGcGM3 tumor-associated ganglioside, was developed, and a clinical trial was conducted to provide a preliminary estimate of the efficacy and safety of racotumomab in patients with advanced non-small-cell lung cancer (15).…”

“…Racotumomab, an antiidiotypic antibody targeting the NeuGcGM3 tumor-associated ganglioside, was developed, and a clinical trial was conducted to provide a preliminary estimate of the efficacy and safety of racotumomab in patients with advanced non-small-cell lung cancer (15). In a separate study, the GD2 antigen, which is highly expressed on neuroblastoma cells in cases of pediatric malignancy with poor prognoses, was targeted through the use of a new human-mouse chimeric anti-idiotypic antibody named ganglidiximab: this antibody was successfully used as a protein vaccine in vivo to induce a GD2-specific humoral immune response (16).…”

Advances in Anti-GD2 Immunotherapy for Treatment of High-risk Neuroblastoma

“…MAb1A7, an anti-Id Ab directed against murine 14G2a (that also recognizes the idiotype of ch14.18), and ganglidiomab, a human/mouse chimeric anti-Id Ab directed against ch14.18 anti-GD2 mAb, have shown similar potential for active immunotherapy against NBL. 62,63 Predicting Response to Immunotherapy with Anti-GD2 mAb Development of human anti-chimeric antibody (HACA), human anti-mouse antibody (HAMA), and human anti-human antibody (HAHA) can be a mechanism of resistance to tumor immunotherapy with anti-GD2 mAb. Patients with strong HACA/HAMA responses following treatment with anti-GD2 mAb show significantly decreased detectable circulating anti-GD2 mAb following subsequent mAb infusions; this was associated with decreased, but not complete abrogation, of ADCC and CDC effector function.…”

Cell death-based treatment of neuroblastoma

“…These designed T cells have a chimeric antigen receptor against GD2, L1-CAM, or ALK, and they have demonstrated safety and no pain toxicity in relapsed NB 248 – 253 . Another similar approach is to use a peptide vaccine, such as ganglidiximab 254 , made from the tumor proteins, to activate T cells against the NB 255 – 257 . These strategies are already in clinical trials and demonstrating high efficiency.…”