The Thomsen-Friedenreich (TF) antigen is a key target for the development of anticancer vaccines, and this ongoing challenge remains relevant due to the poor immunogenicity of the TF antigen.To overcome this challenge, we adopted a bivalent conjugate design which introduced both the TF antigen and Twin Upholstered Headboard the Thomsen-nouveau (Tn) antigen onto the immunologically relevant polysaccharide A1 (PS A1).The immunological results in C57BL/6 mice revealed that the bivalent, Tn-TF-PS A1 conjugate increased the immune response towards the TF antigen as compared to the monovalent TF-PS A1.This phenomenon was first observed with enzyme-linked immunosorbent assay (ELISA) where the bivalent conjugate generated high titers of IgG antibodies where the monovalent conjugate generated an exclusive IgM response.
Fluorescence-activated cell sorting (FACS) analysis also revealed increased binding events to the tumor cell lines MCF-7 and OVCAR-5, which are consistent with the enhanced tumor cell lysis observed in a complement dependent cytotoxicity (CDC) assay.The cytokine profile generated by the bivalent construct revealed increased pro-inflammatory cytokines IL-17 and IFN-γ.This Outdoor Equipment increase in cytokine concentration was matched with an increase in cytokine producing cells as observed by ELISpot.We hypothesized the mechanisms for this phenomenon to involve the macrophage galactose N-acetylgalactosamine specific lectin 2 (MGL2).
This hypothesis was supported by using biotinylated probes and recombinant MGL2 to measure carbohydrate-protein interactions.