Human dendritic cells transfected with amplified MUC1 mRNA stimulate cytotoxic T lymphocyte responses against pancreatic cancer in vitro

Abstract

Background and aim: Mucin (MUC) 1 is an epithelial cell glycoprotein that is aberrantly overexpressed in many adenocarcinomas, including pancreatic cancer (PC), providing an ideal tumor-associated antigen and target for immunotherapy. In this study, we investigated whether the cytotoxic T lymphocytes (CTLs) induced by dendritic cells (DCs) transfected with amplified MUC1 mRNA could respond against PC in vitro.

Methods: Amplified mRNA encoding MUC1 were transfected into DCs using electroporation with an optimized setting and the MUC1 expression were evaluated by quantitative real-time polymerase chain reaction and Western blot. The MUC1 specific CTL responses were measured using the standard chromium 51 (51Cr)-release assays and the interferon-γ release assay.

Results: Dendritic cells could be transfected with amplified MUC1 mRNA efficiently. The transfected DCs were remarkably effective in stimulating MUC1-specific CTL responses in vitro. The function of MUC1 specific CTLs, induced by MUC1 mRNA-transfected DCs, was restricted by major histocompatibility complex (MHC) class I antigen presentation.

Conclusion: The CTL responses stimulated by DCs transfected with MUC1 mRNA could only recognize and lyse HLA-A2+/MUC1+ PC and other target cells under restriction by MHC class I-specific antigen presentation, providing a preclinical rationale for using MUC1 as a target structure for immunotherapeutic strategies against PC.