Anti-CD3/Anti-Epidermal Growth Factor Receptor-Bispecific Antibody Retargeting of Lymphocytes against Human Neoplastic Keratinocytes in an Autologous Organotypic Culture Model

摘要

Local cellular immune defects have been described in severaltumors including human papillomavirus (HPV)-associated cervical cancer.This observation suggests the potential therapeutic benefit of immunemanipulations that restore cellular immunity. Here, weevaluated the ability of bispecific monoclonal antibodies (bimAbs) toredirect T cells against keratinocytes transformed invitro by HPV in an autologous three-dimensional culture model(organotypic cultures). The epidermal growth factor receptor (EGFR) waschosen as target for an anti-CD3/anti-EGFR bimAb because it isoverexpressed in many malignant epithelial lesions and only weaklyexpressed in the basal layers of normal squamous epithelium.Interestingly, in organotypic cultures, the pattern ofexpression of EGFR was similar to that observed in vivo.The ability of T cells retargeted by CD3/EGFR bimAb to lyseHPV-transformed cell lines was confirmed in monolayer cultures. Inautologous organotypic cultures, an increase in apoptoticHPV+ keratinocytes and a significant decrease in thethickness of HPV+ organotypic cultures were observed whenactivated lymphocytes and bimAbs were added to the cultures,whereas organotypic cultures of normal keratinocytes were notsignificantly affected. These data were similar to those obtained inthe allogeneic model. These results suggest the potential usefulness ofCD3-EGFR bimAb-retargeted lymphocytes in immunotherapeutic protocolsfor malignant epithelial lesions.