A preclinical study comparing approaches for augmenting the immunogenicity of a heptavalent KLH-conjugate vaccine against XXXX .

@entity15670 @entity6496 @entity5 @entity3857 @entity9221 @entity786 @entity35650 @entity2355 @entity17080 @entity4386
Previously using a series of monovalent vaccines, we demonstrated that the optimal method for inducing an antibody response against @entity5 cell-surface antigens is covalent conjugation of the antigens to keyhole limpet hemocyanin (KLH) and the use of a @entity3857 adjuvant. We have prepared a heptavalent-KLH conjugate vaccine containing the seven epithelial @entity5 antigens GM2, Globo H, Lewis(y), TF(c), @entity15670 , @entity9221 (c), and glycosylated @entity2355 . 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 @entity786 or anti- @entity4386 MAb), different @entity3857 adjuvants at various doses ( @entity17080 and @entity35650 ), and different methods of formulation (lyophilization and use of @entity6496 80). We find that: (1). Immunization with the heptavalent-KLH conjugate plus @entity35650 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), @entity9221 (640/5120), TF (320/10240), @entity2355 (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 @entity5 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 @entity6496 80 had any impact on antibody titers. (4). An optimal dose of @entity3857 adjuvant, @entity17080 (50 microg) or @entity35650 (1000 microg), is required for optimal antibody titers. This heptavalent vaccine is sufficiently optimized for testing in the clinic.
