Identification of an HLA-A*0201-restrictive CTL epitope from MUC4 for applicable vaccine therapy

Abstract

Recent research has indicated that MUC4 plays an important role in the development of many tumors and may prove useful as a novel cancer immunotherapy target. We aimed to identify HLA-A*0201-restrictive cytotoxic T lymphocyte (CTL) epitopes of the cancer-associated antigen MUC4. The MUC4 sequence was scanned for immunogenic peptides using HLA-binding prediction software. Dendritic cells (DCs) from peripheral blood mononuclear cells (PBMCs) were induced by cytokines. Five possible CTL epitopes were selected by software analysis, synthesized, and used to pulse mature DCs. The CD8⁺ T cells from PBMCs from an HLA-A*0201 healthy donor were stimulated with autologous MUC4-peptide-loaded DCs and expanded in vitro. T cell activation was assessed by ELISPOT, and cytotoxicity was determined by ⁵¹chromium (⁵¹Cr)-release assays. Our results show that CTLs induced by peptide P01204 could lyse T2 cells pulsed with peptide P01204 and HCT-116 cells (MUC4⁺, HLA-A2⁺). Compared with a control peptide, P01204 increased the number of IFN-γ producing T cells. Overall, these results suggest that P01204 is a novel HLA-A*0201-restrictive CTL epitope of the cancer-associated antigen MUC4. This will provide a foundation for the development of tumor-specific peptide vaccines.

Keywords::

• Mucin4
• peptide
• dendritic cell
• cytotoxic T lymphocyte
• vaccine

Acknowledgments

This work was supported by grants from the National Nature Science Foundation of China (No. 30500492 and No. 30471691).

Declaration of interest: The authors report no conflicts of interest. The authors alone are responsible for the content and writing of the paper.