Nanobody-Antigen Conjugates Elicit HPV-Specific Antitumor Immune Responses

High-risk human papillomavirus-associated cancers express viral oncoproteins (e.g., E6 and E7) that induce and maintain the malignant phenotype. The viral origin of these proteins makes them attractive targets for development of a therapeutic vaccine. Camelid-derived single-domain antibody fragments (nanobodies or VHHs) that recognize cell surface proteins on antigen-presenting cells (APC) can serve as targeted delivery vehicles for antigens attached to them. Such VHHs were shown to induce CD4⁺ and CD8⁺ T-cell responses against model antigens conjugated to them via sortase, but antitumor responses had not yet been investigated. Here, we tested the ability of an anti-CD11b VHH (VHH_CD11b) to target APCs and serve as the basis for a therapeutic vaccine to induce CD8⁺ T-cell responses against HPV⁺ tumors. Mice immunized with VHH_CD11b conjugated to an H-2D^b-restricted immunodominant E7 epitope (E7_49-57) had more E7-specific CD8⁺ T cells compared with those immunized with E7_49-57 peptide alone. These CD8⁺ T cells acted prophylactically and conferred protection against a subsequent challenge with HPV E7-expressing tumor cells. In a therapeutic setting, VHH_CD11b-E7_49-57 vaccination resulted in greater numbers of CD8⁺ tumor-infiltrating lymphocytes compared with mice receiving E7_49-57 peptide alone in HPV⁺ tumor-bearing mice, as measured by in vivo noninvasive VHH-based immune-positron emission tomography (immunoPET), which correlated with tumor regression and survival outcome. Together, these results demonstrate that VHHs can serve as a therapeutic cancer vaccine platform for HPV-induced cancers. Cancer Immunol Res; 6(7); 870-80. ©2018 AACR.