The effect of the modification of ceramide on the carbohydrate antigenicity of gangliosides

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  • ガングリオシド糖鎖の抗原性に対するセラミド修飾体の影響

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A murine monoclonal antibody （IgMκ）, termed 3D10, was established by fusion of P3X63Ag8.653 mouse myeloma cells with spleen cells from C3H/HeN mouse immunized with gliomarelated O-acetyl GM3 （NeuAc） having 3-O-acetyl ceramide. The antigens recognized by 3D10 were elucidated through enzyme-linked immunosorbent assay （ELISA） and immunostaining on thin-layer chromatography （TLC-immunostaining）. 3D10 reacts with 3-O-acetyl GM3 （NeuAc） in a strong manner than non-acetylated GM3 （NeuAc）, but not with 3-O-acetyl GM3 （NeuGc）, GM3 （NeuGc）, GM1 （NeuAc）, GM2 （NeuAc）, GD3 （NeuAc）, lactosylceramide or glucosylceramide. Furthermore, the reactivity of 3-O-acetyl GM3 （NeuAc） and non-acetylated GM3 （NeuAc） with the monoclonal antibody M2590, which recognizes terminal NeuAc α2?3Galβ1?4Glc （or GlcNAc） residue as a melanoma antigen, was studied through ELISA, TLC-immunostaining and liposome immune lysis assay （LILA）. 3-O-acetyl GM3 （NeuAc） showed a lower reactivity than GM3 in these three kinds of assay systems, and this was contrastingly different from the reactivity of 3D10 when tested with the same assay systems. Interestingly, a mixture of 3-O-acetyl GM3 （NeuAc） and non- acetylated GM3 （NeuAc） showed the almost same threshold as GM3 and the nearly same maximum reactivity as 3-O-acetyl GM3 （NeuAc） in LILA. Antibody response after immunization with the 3-O-acetyl GM3 （NeuAc）, GM3 （NeuAc）, a mixture of GM3 and 3-O-acetyl GM3 （65:35, mol:mol） in the mouse was studied. The liposome containing each ganglioside was injected intraperitoneally and serum antibody response was assayed by ELISA. GM3 induced a low titer anti-GM3 antibody response but an extremely low anti-3-O-?acetyl GM3 titer. 3-O-acetyl GM3 and a mixture of 3-O-acetyl GM3 and GM3 only minimally induced the elevation of serum anti-3-O-acetyl GM3 or anti-GM3 antibody response. These findings suggest that the existence of 3-O-acetyl GM3 could suppress the antibody response in mice.

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