Generation and Characterization of an Anti-Idiotype Monoclonal Antibody Related to GM3(NeuGc) Ganglioside

Abstract

The 14F7 monoclonal antibody (MAb), IgG1 isotype, which reacts specifically to GM3(NeuGc) ganglioside induced a specific IgG anti-idiotypic antibody (Ab2) response in syngeneic mice when it was administered coupled with KLH and in the presence of Freund's adjuvant. Spleen cells from these mice were used in somatic-cell hybridization experiments using the murine myeloma cell line P3-X63-Ag8 653 as fusion partner. An IgG1 Ab2 MAb was selected. This Ab2 MAb, called 4G9, was able to block the binding of 14F7 MAb to GM3(NeuGc) ganglioside and developed a strong IgG anti-anti-idiotypic antibody (Ab3) response, when injected into syngeneic mice. These Ab3 antibodies were characterized to bear 14F7 MAb idiotopes, but did not have the same specificity as 14F7 MAb. In the other hand, a very specific anti-NeuGc-containing ganglioside response was generated in chickens immunized with this Ab2 MAb, thus behaving, in this species as an "internal image" antibody.

Generation of anti-Neu-glycolyl-ganglioside antibodies by immunization with an anti-idiotype monoclonal antibody: A self versus non-self-matter

“…In fact, we have generated not only IgM Ab1 mAbs, as P3 (Va´zquez et al, 1995a), but also IgG1 mAb, as 14F7, very specific to GM 3 (NeuGc) ganglioside (Carr et al, 2000). But anti-idiotype antibodies to 14F7 mAb also failed to break tolerance to NeuGc gangliosides in mice, while inducing an antigen-specific-antibody response in chickens (Rodrı´guez et al, 2003).…”

Anti-ganglioside antibody-induced tumor cell death by loss of membrane integrity

“…This mAb is able to block the binding of 14F7 mAb to the ganglioside (18). The anti-idiotype 4G9 mAb impaired 14F7-induced NGcGM3-mediated cell death in a dose-dependent manner (Fig.…”

“…11), 14F7 anti-idiotypic 4G9 mAb (murine IgG1, n; ref. 18), iorC5 mAb (murine IgG1, n) that recognizes a glycoprotein expressed in colorectal cancer cells (19) and chimeric P3 (human IgG1, n), an anti-NGc-containing gangliosides and sulfatides mAb (20), were purified by Protein A Affinity Chromatography (Pharmacia) and analyzed by SDS-PAGE under reducing conditions. F(ab ¶) 2 and Fab fragments antibodies were obtained using a standard procedure described previously (21).…”

Insights into the immunogenetic basis of two ganglioside-associated idiotypic networks

“…Both MAbs were able to react with antigens expressed in human melanoma and breast tumors (Va´zquez et al, 1995a;Carr et al, 2000;Alfonso et al, 2002;Oliva et al, 2006). P3 and 14F7 MAbs, coupled to a carrier protein and in the presence of Freund's adjuvant, induced anti-idiotypic responses in syngeneic mice, and specific Ab2 MAbs were generated and characterized Rodrı´guez et al, 2003). In addition, P3 MAb has the uncommon ability to induce an IgG anti-idiotypic antibody response in BALB/c mice even when administered in saline .…”

“…In addition, P3 MAb has the uncommon ability to induce an IgG anti-idiotypic antibody response in BALB/c mice even when administered in saline . On the other hand, 1E10 (anti-P3) and 4G9 (anti-14F7) Ab2 MAbs were not able to induce anti-GM3(NeuGc) ganglioside Ab3 antibodies in syngeneic model, but induced an antigen-positive humoral response in chickens (Rodrı´guez et al, 2003;Herna´ndez et al, 2005) and, in the case of 1E10 MAb, also in humans (Alfonso et al, 2002;Dı´az et al, 2003), where N-glycolyl-gangliosides are non-self-antigens.…”