Assessing vaccine potency using TCRmimic antibodies

• Published in: Vaccine 2008-06, Vol.26 (25), p.3092-3102
• Main Authors: Neethling, Francisca A, Ramakrishna, Venky, Keler, Tibor, Buchli, Rico, Woodburn, Tito, Weidanz, Jon A
• Format: Article
• Language: English
• Subjects: Allergy and Immunology; Antibodies, Monoclonal - administration & dosage; Antibodies, Monoclonal - immunology; Antibody Specificity; Antigens; Antigens, Neoplasm - immunology; Cancer; Cancer Vaccines - administration & dosage; Cancer Vaccines - immunology; Cell Line; Cytokines; Cytotoxic T lymphocyte epitopes; Cytotoxicity; Dendritic Cells - immunology; Epitopes - immunology; Epitopes - metabolism; HLA-A2 Antigen - genetics; HLA-A2 Antigen - immunology; Human leukocyte antigens; Humans; Immune system; Lymphocytes; Monoclonal antibody; Peptides; Proteins; Receptors, Antigen, T-Cell - immunology; Studies; T cell receptor mimics; T cell receptors; T-Lymphocytes, Cytotoxic - immunology; Tumor-associated antigens; Vaccine potency assays; Vaccines
• ISSN: 0264-410X; 1873-2518
• DOI: 10.1016/j.vaccine.2008.02.025

Summary Dendritic cells (DCs) are highly specialized antigen-presenting cells of the immune system that are efficient at presenting peptide-antigen for the activation of T cells and are often the cell type of choice for vaccine targeting by virtue of high expression levels of MHC and costimulatory molecules. Since the level of peptide–MHC complex significantly influences stimulation of T cells, a proof-of-concept potency assay was developed to directly examine the presentation and density of MHC class I peptides derived from the processing of a model tumor antigen, (hCGβ), on the surface of DCs. In this study we first generated antibodies (TCR mimics or TCRm) to two peptide-HLA-A*0201 epitopes derived from hCGβ designated as TMT (40–48) and GVL (47–55). Characterization of each TCRm by ELISA and flow cytometric analysis, demonstrated specific binding to soluble recombinant HLA-A2 protein and HLA-A2.1+ T2 cells loaded with relevant peptide. TCRm reactive against the TMT and GVL epitopes blocked granzyme-B production by peptide-specific cytotoxic T lymphocytes (CTL) lines further supporting their recognition specificity. For the assessment of antigen presentation function, human immature monocyte-derived DCs (iDCs) were treated with the mannose receptor targeting vaccine, B11-hCGβ and matured with Poly I:C. The TMT and GVL epitope reactive CTL lines responded to vaccine-treated but not vehicle-treated mature DCs (mDCs) with TMT and GVL TCRm specifically blocking IFN-γ production. The TCRm were then used to directly confirm specific peptide–MHC complexes on mDCs. TCRm staining of vaccine-treated mDCs showed detection of the TMT and GVL peptide-HLA-A2 complexes. These findings demonstrate that TCRms may be important tools for determining the potency of DC-based vaccines.