Production of monoclonal antibodies directed to Hanganutziu-Deicher active gangliosides, N-glycolylneuraminic acid-containing gangliosides.
We have established three kinds of monoclonal antibodies against gangliosides containing N-glycolylneuraminic acid (NeuGc) by immunization of BALB/c mice with the purified gangliosides inserted into liposomes comprising Salmonella minnesota R595 lipopolysaccharides, and fusion of spleen cells with a mouse myeloma cell line. One monoclonal antibody, SHS-1, which was generated by immunizing mice with purified i-active ganglioside(NeuGc), reacted specifically with the i-active ganglioside(NeuGc) used as an immunogen. Structurally related gangliosides, such as GM3(NeuGc), sialosylparagloboside (SPG) (NeuGc), or I-active ganglioside(NeuGc), corresponding gangliosides [GM3 containing N-acetylneuraminic acid (NeuAc), SPG(NeuAc), i-active ganglioside(NeuAc), and I-active ganglioside(NeuAc)], other gangliosides, or neutral glycosphingolipid (GSL) were not recognized by the monoclonal antibody. These findings indicate that the SHS-1 monoclonal antibody may be specific for NeuGc-containing i-active ganglioside. On the other hand, the other two monoclonal antibodies, MSG-1 and SPS-20, which were generated by immunizing mice with purified ganglioside GM3(NeuGc) and SPG(NeuGc), respectively, showed cross-reactivity to structurally related gangliosides. The MSG-1 monoclonal antibody exhibited reactivity to ganglioside GM3(NeuAc). The SPS-20 monoclonal antibody also cross-reacted with SPG(NeuAc), i-active ganglioside(NeuGc), and i-active ganglioside(NeuAc). Neither MSG-1 nor SPS-20 reacted with corresponding gangliosides, other gangliosides, or neutral GSLs tested. Using the SHS-1 antibody specific for i-active ganglioside(NeuGc), we studied the expression of NeuGc-containing antigen in human colon cancer tissue. An NeuGc-containing glycoconjugate was detected in the colon cancer tissue.
Watarai
S
S
Department of Cell Chemistry, Okayama University Medical School.
Kushi
Y
Y
Shigeto
R
R
Misawa
N
N
Eishi
Y
Y
Handa
S
S
Yasuda
T
T
D000818:Animals; D000910:Antibodies, Heterophile / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D000917:Antibody Formation; D000918:Antibody Specificity; D015478:Antigens, Heterophile / Q000276:immunology*; D000951:Antigens, Neoplasm / Q000032:analysis / Q000737:chemistry / Q000276:immunology; D002240:Carbohydrate Sequence; D003110:Colonic Neoplasms / Q000737:chemistry / Q000276:immunology; D005732:Gangliosides / Q000737:chemistry / Q000276:immunology*; D005915:Globosides / Q000276:immunology; D006801:Humans; D007150:Immunohistochemistry; D008081:Liposomes / Q000737:chemistry / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009438:Neuraminic Acids / Q000276:immunology
D000910:Antibodies, Heterophile; D000911:Antibodies, Monoclonal; D015478:Antigens, Heterophile; D000951:Antigens, Neoplasm; D005732:Gangliosides; D005915:Globosides; D008081:Liposomes; D009438:Neuraminic Acids; C014812:sialosylparagloboside