title: A preclinical study comparing approaches for augmenting the immunogenicity of a heptavalent KLH-conjugate vaccine against epithelial cancers.
abstract: Previously using a series of monovalent vaccines, we demonstrated that the optimal method for inducing an antibody response against cancer cell-surface antigens is covalent conjugation of the antigens to keyhole limpet hemocyanin (KLH) and the use of a saponin adjuvant. We have prepared a heptavalent-KLH conjugate vaccine containing the seven epithelial cancer antigens GM2, Globo H, Lewis(y), TF(c), Tn(c), STn(c), and glycosylated MUC1. In preparation for testing this vaccine in the clinic, we tested the impact on antibody induction of administering the individual conjugates plus adjuvant compared with a mixture of the seven conjugates plus adjuvant, and of several variables thought to augment immunogenicity. These include approaches for decreasing suppressor cell activity or increasing helper T-lymphocyte activity (low dose cyclophosphamide or anti-CTLA-4 MAb), different saponin adjuvants at various doses (QS-21 and GPI-0100), and different methods of formulation (lyophilization and use of polysorbate 80). We find that: (1). Immunization with the heptavalent-KLH conjugate plus GPI-0100 vaccine induces antibodies against the seven antigens of comparable titer to those induced by the individual-KLH conjugate vaccines, high titers of antibodies against Tn (median ELISA titer IgM/IgG 320/10240), STn (640/5120), TF (320/10240), MUC1 (80/20480), and globo H (640/40); while lower titers of antibodies against Lewis(y)()(160/0) and only occasional antibodies against GM2 are induced. (2). These antibodies reacted with the purified synthetic antigens by ELISA, and with naturally expressed antigens on the cancer cell surface by FACS. (3). None of the approaches for further altering the suppressor cell/helper T-cell balance nor changes to the standard formulation by lyophilization or use of polysorbate 80 had any impact on antibody titers. (4). An optimal dose of saponin adjuvant, QS-21 (50 microg) or GPI-0100 (1000 microg), is required for optimal antibody titers. This heptavalent vaccine is sufficiently optimized for testing in the clinic.
authors.affiliation: Laboratory of Tumor Vaccinology, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, New York, NY 10021, USA. ragupatg@mskcc.org
authors.forename: Govind
authors.initials: G
authors.lastname: Ragupathi
authors.forename: Fusataka
authors.initials: F
authors.lastname: Koide
authors.forename: Natarajan
authors.initials: N
authors.lastname: Sathyan
authors.forename: Ella
authors.initials: E
authors.lastname: Kagan
authors.forename: Maria
authors.initials: M
authors.lastname: Spassova
authors.forename: William
authors.initials: W
authors.lastname: Bornmann
authors.forename: Polly
authors.initials: P
authors.lastname: Gregor
authors.forename: Celso A
authors.initials: CA
authors.lastname: Reis
authors.forename: Henrik
authors.initials: H
authors.lastname: Clausen
authors.forename: Samuel J
authors.initials: SJ
authors.lastname: Danishefsky
authors.forename: Philip O
authors.initials: PO
authors.lastname: Livingston
mesh_terms: D000818:Animals; D000912:Antibodies, Neoplasm / Q000096:biosynthesis*; D000917:Antibody Formation; D000951:Antigens, Neoplasm / Q000276:immunology*; D019496:Cancer Vaccines / Q000276:immunology*; D004353:Drug Evaluation, Preclinical; D005260:Female; D005732:Gangliosides / Q000008:administration & dosage / Q000276:immunology; D006433:Hemocyanins / Q000276:immunology*; D007114:Immunization; D007074:Immunoglobulin G / Q000276:immunology / Q000378:metabolism; D007075:Immunoglobulin M / Q000276:immunology / Q000378:metabolism; D007983:Lewis Blood Group Antigens / Q000008:administration & dosage / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D018396:Mucin-1 / Q000008:administration & dosage / Q000276:immunology; D009375:Neoplasms, Glandular and Epithelial / Q000276:immunology* / Q000473:pathology; D010446:Peptide Fragments / Q000008:administration & dosage / Q000276:immunology; D012503:Saponins / Q000008:administration & dosage / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology; D014611:Vaccination; D018074:Vaccines, Conjugate / Q000276:immunology
chemical_list: D000912:Antibodies, Neoplasm; D000951:Antigens, Neoplasm; D019496:Cancer Vaccines; D005732:Gangliosides; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D007983:Lewis Blood Group Antigens; C094162:Lewis Y antigen; C116055:MUC1 tandem repeat peptide; D018396:Mucin-1; D010446:Peptide Fragments; D012503:Saponins; D018074:Vaccines, Conjugate; C078785:saponin QA-21V1; D006433:Hemocyanins; C032808:keyhole-limpet hemocyanin