title: GD3 vaccines for melanoma: superior immunogenicity of keyhole limpet hemocyanin conjugate vaccines.
abstract: Cell surface gangliosides show altered patterns of expression as a consequence of malignant transformation and have therefore been of interest as potential targets for immunotherapy, including vaccine construction. One obstacle has been that some of the gangliosides that are overexpressed in human cancers are poorly immunogenic in humans. A case in point is GD3, a prominent ganglioside of human malignant melanoma. Using an approach that has been effective in the construction of bacterial carbohydrate vaccines, we have succeeded in increasing the immunogenicity of GD3 in the mouse by conjugating the ganglioside with immunogenic carriers. Several conjugation methods were used. The optimal procedure involved ozone cleavage of the double bond of GD3 in the ceramide backbone, introducing an aldehyde group, and coupling to aminolysyl groups of proteins by reductive amination. Conjugates were constructed with a synthetic multiple antigenic peptide expressing repeats of a malarial T-cell epitope, outer membrane proteins of Neisseria meningitidis, cationized bovine serum albumin, keyhole limpet hemocyanin, and polylysine. Mice immunized with these conjugates showed a stronger antibody response to GD3 than mice immunized with unconjugated GD3. The strongest response was observed in mice immunized with the keyhole limpet hemocyanin conjugate of the GD3 aldehyde derivative and the adjuvant QS-21. These mice showed not only a long-lasting high-titer IgM response but also a consistent high-titer IgG response (predominantly IgG1), indicating recruitment of T-cell help, although the titers of IgM and IgG antibodies following booster immunizations were not as high as they are in the response to classical T-cell-dependent antigens. This method is applicable to other gangliosides, and it may be useful in the construction of immunogenic ganglioside vaccines for the immunotherapy of human cancers expressing gangliosides on their cell surface.
authors.affiliation: Immunology Program, Memorial Sloan-Kettering Cancer Center, New York, New York 10021.
authors.forename: F
authors.initials: F
authors.lastname: Helling
authors.forename: A
authors.initials: A
authors.lastname: Shang
authors.forename: M
authors.initials: M
authors.lastname: Calves
authors.forename: S
authors.initials: S
authors.lastname: Zhang
authors.forename: S
authors.initials: S
authors.lastname: Ren
authors.forename: R K
authors.initials: RK
authors.lastname: Yu
authors.forename: H F
authors.initials: HF
authors.lastname: Oettgen
authors.forename: P O
authors.initials: PO
authors.lastname: Livingston
mesh_terms: D000276:Adjuvants, Immunologic; D000818:Animals; D000912:Antibodies, Neoplasm / Q000276:immunology; D005260:Female; D005732:Gangliosides / Q000737:chemistry / Q000276:immunology*; D006433:Hemocyanins / Q000276:immunology; D000922:Immunotoxins / Q000276:immunology*; D008545:Melanoma / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D009844:Oligosaccharides / Q000276:immunology*; D012503:Saponins / Q000276:immunology; D014612:Vaccines / Q000737:chemistry / Q000276:immunology*
chemical_list: D000276:Adjuvants, Immunologic; D000912:Antibodies, Neoplasm; D005732:Gangliosides; D000922:Immunotoxins; D009844:Oligosaccharides; D012503:Saponins; D014612:Vaccines; C078785:saponin QA-21V1; C026226:ganglioside, GD3; D006433:Hemocyanins; C032808:keyhole-limpet hemocyanin