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Determination of cellularly processed HLA-A2402-restricted novel CTL epitopes derived from two cancer germ line
Yoshihiro Miyahara1, Hiroaki Naota, Lijie Wang
1Second Department of Internal Medicine and Department of Otorhinolaryngology, Mie University School of Medicine, Mie, Japan.
Purpose:
For identification of CTL epitopes useful for cancer vaccines, it is crucial to determine whether cognate epitopes are presented on the cell surface of target cancer cells through natural processing of endogenous proteins. For this purpose, we tried to use the cellular machinery of both mice and human to define naturally processed CTL epitopes derived from two "cancer germ line" genes, MAGE-A4 and SAGE.
Experimental Design:
We vaccinated newly produced HLA-A2402 transgenic mice with DNA plasmids encoding target antigens. Following screening of synthesized peptides by splenic CD8(+) T cells of vaccinated mice, we selected candidate epitopes bound to HLA-A2402. We then examined whether human CD8(+) T cells sensitized with autologous CD4(+) PHA blasts transduced by mRNA for the cognate antigens could react with these selected peptides in an HLA-A2402-restricted manner.
Results:
After DNA vaccination, murine CD8(+) T cells recognizing MAGE-A4(143-151) or SAGE(715-723) in an HLA-A2402-restricted manner became detectable. Human CTLs specific for these two peptides were generated after sensitization of HLA-A2402-positive CD8(+) T cells with autologous CD4(+) PHA blasts transduced with respective mRNA. CTL clones were cytotoxic toward tumor cell lines expressing HLA-A2402 and cognate genes. Taken together, these CTL epitopes defined in HLA-A24 transgenic mice are also processed and expressed with HLA-A2402 in human cells. The presence of SAGE(715-723)-specific precursors was observed in HLA-A2402-positive healthy individuals.
Conclusions:
Two novel HLA-A2402-restricted CTL epitopes, MAGE-A4(143-151) and SAGE(715-723), were identified. Our approach assisted by cellular machinery of both mice and human could be widely applicable to identify naturally processed CTL epitopes.
Insights
Researchers identified two novel cancer-targeting epitopes, MAGE-A4(143-151) and SAGE(715-723), crucial for developing effective cancer vaccines. This approach uses both mouse and human cellular machinery for natural epitope processing and presentation.
Area of Science:
- Immunology
- Oncology
- Vaccine Development
Background:
- Identifying naturally processed cytotoxic T lymphocyte (CTL) epitopes presented on cancer cells is vital for effective cancer vaccine design.
- Cancer germline genes like MAGE-A4 and SAGE are key targets for cancer immunotherapy.
Purpose of the Study:
- To define naturally processed CTL epitopes derived from MAGE-A4 and SAGE using both murine and human cellular systems.
- To validate these epitopes for their potential use in HLA-A2402-restricted cancer vaccines.
Main Methods:
- DNA vaccination of HLA-A2402 transgenic mice with MAGE-A4 and SAGE antigens.
- Screening of synthesized peptides using splenic CD8(+) T cells from vaccinated mice.
- Sensitization of human CD8(+) T cells with mRNA-transduced autologous CD4(+) PHA blasts to assess peptide reactivity.
Main Results:
- Murine CD8(+) T cells recognizing MAGE-A4(143-151) and SAGE(715-723) in an HLA-A2402-restricted manner were detected post-vaccination.
- Human CTLs specific for these peptides were generated and demonstrated cytotoxicity against relevant tumor cell lines.
- The identified CTL epitopes are naturally processed and presented on cancer cells in an HLA-A2402-restricted manner, with SAGE(715-723)-specific precursors found in healthy individuals.
Conclusions:
- Two novel HLA-A2402-restricted CTL epitopes, MAGE-A4(143-151) and SAGE(715-723), were successfully identified.
- The combined use of mouse and human cellular machinery provides a broadly applicable method for discovering naturally processed CTL epitopes for cancer vaccines.
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