Polymeric Structure and Host Toll-like Receptor 4 Dictate Immunogenicity of NY-ESO-1 Antigen in Vivo

• Published in: The Journal of biological chemistry 2011-10, Vol.286 (43), p.37077-37084
• Main Authors: Liu, Yanan, Tian, Xiaoli, Leitner, Wolfgang W., Aldridge, Michael E., Zheng, Junying, Yu, Zhiya, Restifo, Nicholas P., Weiss, Richard, Scheiblhofer, Sandra, Xie, Chong, Sun, Ren, Cheng, Genhong, Zeng, Gang
• Format: Article
• Language: English
• Subjects: Animals; Antibodies, Neoplasm - immunology; Antibody Formation - genetics; Antibody Formation - immunology; Antigen; Antigens, Neoplasm - genetics; Antigens, Neoplasm - immunology; Calreticulin; Cancer Vaccines - genetics; Cancer Vaccines - immunology; Cell Line, Tumor; Cell Surface Receptor; Damage-associated Molecular Pattern; Dendritic Cells; Dendritic Cells - immunology; HEK293 Cells; Humans; Immunology; Male; Membrane Proteins - genetics; Membrane Proteins - immunology; Mice; Mice, Knockout; Protein Binding - genetics; Protein Binding - immunology; Protein Multimerization - genetics; Protein Multimerization - immunology; Toll-Like Receptor 4 - genetics; Toll-Like Receptor 4 - immunology; Toll-like Receptors (TLR); Tumor Immunology; Tumor-associated Antigen; Vaccine Development; Vaccines, DNA - genetics; Vaccines, DNA - immunology
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M111.280123

In search of intrinsic factors that contribute to the distinctively strong immunogenicity of a non-mutated cancer/testis antigen, we found that NY-ESO-1 forms polymeric structures through disulfide bonds. NY-ESO-1 binding to immature dendritic cells was dependent on its polymeric structure and involved Toll-like receptor-4 (TLR4) on the surface of immature dendritic cells in mouse and human. Gene gun-delivered plasmid encoding the wild-type NY-ESO-1 readily induced T cell-dependent antibody (Ab) responses in wild-type C57BL/10 mice but not TLR4-knock-out C57BL/10ScNJ mice. Disrupting polymeric structures of NY-ESO-1 by cysteine-to-serine (Cys-to-Ser) substitutions lead to diminished immunogenicity and altered TLR4-dependence in the induced Ab response. To demonstrate its adjuvant effect, NY-ESO-1 was fused with a major mugwort pollen allergen Art v 1 and a tumor-associated antigen, carbonic anhydrase 9. Plasmid DNA vaccines encoding the fusion genes generated robust immune responses against otherwise non-immunogenic targets in mice. Polymeric structure and TLR4 may play important roles in rendering NY-ESO-1 immunogenic and thus serve as a potent molecular adjuvant. NY-ESO-1 thus represents the first example of a cancer/testis antigen that is a also damage-associated molecular pattern.