Immunization of mice with fucosyl-GM1 conjugated with keyhole limpet hemocyanin results in antibodies against human small-cell lung cancer cells.
Fucosyl-GM1 (Fuc-GM1) [Fucalpha1 --> 2Galbeta1 --> 3GalNAcbeta1 --> 4(NeuAcalpha2-3)Galbeta1 --> 4Glcbeta1 --> O-Cer] is a small-cell-lung-cancer (SCLC)-associated ganglioside initially defined by the murine monoclonal antibody F12. On the basis of its known distribution, Fuc-GM1 is a potential target for active immunotherapy in SCLC patients. Fuc-GM1 has been extracted and purified from bovine thyroid. The immunogenicity of Fuc-GM1 was tested in mice either alone, mixed with carrier protein keyhole limpet hemocyanin (KLH) or covalently linked with KLH, plus immunological adjuvant QS-21. The Fuc-GM1-KLH conjugate plus QS-21 adjuvant was found to be optimal. It induced consistent IgM and IgG enzyme-linked immunosorbent assay (ELISA) titers against Fuc-GM1. These antibodies were strongly reactive with the strongly Fuc-GM1-positive rat hepatoma cell line H4-II-E, and they were moderately reactive with the moderately positive human SCLC cell line H146 by flow cytometry and complement-mediated lysis. Both ELISA and fluorescence-activated cell sorting (FACS) reactions were inhibited with Fuc-GM1or H4-II-E but not with the structurally related ganglioside GM1 or Fuc-GM1-negative colon cancer cell line LS-C. On the basis of these results, a vaccine comprising Fuc-GM1-KLH plus QS-21 is being prepared for testing in patients with SCLC.
Cappello
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S
Clinical Immunology Service, Department of Medicine, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, New York, NY 10021, USA.
Liu
N X
NX
Musselli
C
C
Brezicka
F T
FT
Livingston
P O
PO
Ragupathi
G
G
D000276:Adjuvants, Immunologic; D000818:Animals; D000912:Antibodies, Neoplasm / Q000096:biosynthesis* / Q000276:immunology; D019496:Cancer Vaccines / Q000276:immunology*; D018288:Carcinoma, Small Cell / Q000276:immunology / Q000628:therapy*; D002417:Cattle; D003110:Colonic Neoplasms / Q000473:pathology; D003165:Complement System Proteins / Q000276:immunology; D003602:Cytotoxicity, Immunologic; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005677:G(M1) Ganglioside / Q000031:analogs & derivatives* / Q000276:immunology; D006801:Humans; D007114:Immunization; D008114:Liver Neoplasms, Experimental / Q000473:pathology; D008175:Lung Neoplasms / Q000276:immunology / Q000628:therapy*; D051379:Mice; D009928:Organ Specificity; D051381:Rats; D012503:Saponins; D014407:Tumor Cells, Cultured
D000276:Adjuvants, Immunologic; D000912:Antibodies, Neoplasm; D019496:Cancer Vaccines; D012503:Saponins; D005677:G(M1) Ganglioside; C078785:saponin QA-21V1; C039123:fucosyl GM1 ganglioside; D003165:Complement System Proteins