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Updated: Aug 15, 2026
Isolation and Flow Cytometric Analysis of Glioma-infiltrating Peripheral Blood Mononuclear Cells
Published on: November 28, 2015
EphA2 as a glioma-associated antigen: a novel target for glioma vaccines
Manabu Hatano1, Junichi Eguchi, Tomohide Tatsumi
1Department of Neurological Surgery, University of Pittsburgh School of Medicine and University of Pittsburgh Cancer Institute, Pittsburgh, PA 15213, USA.
Abstract:
EphA2 is a receptor tyrosine kinase and is frequently overexpressed in a wide array of advanced cancers. We demonstrate in the current study that the EphA2 protein is restrictedly expressed in primary glioblastoma multiforme and anaplastic astrocytoma tissues in comparison to normal brain tissues. To evaluate the possibility of targeting EphA2 in glioma vaccine strategies, we stimulated human leukocyte antigen (HLA) A2+ peripheral blood mononuclear cells (PBMCs) obtained from healthy donors and glioma patients with autologous dendritic cells (DCs) loaded with synthetic EphA2883-891 peptide (TLADFDPRV), which has previously been reported to induce interferon-gamma in HLA-A2+ PBMCs. Stimulated PBMCs demonstrated antigen-specific cytotoxic T lymphocyte (CTL) responses as detected by specific lysis of T2 cells loaded with the EphA2883 peptide as well as HLA-A2+ glioma cells, SNB19 and U251, that express EphA2. Furthermore, in vivo immunization of HLA-A2 transgenic HHD mice with the EphA2883-891 peptide resulted in the development of an epitope-specific CTL response in splenocytes, despite the fact that EphA2883-891 is an autoantigen in these mice. Taken together, these data suggest that EphA2883-891 may be an attractive antigen epitope for molecularly targeted glioma vaccines.
Insights
EphA2 receptor tyrosine kinase is overexpressed in cancers. This study shows EphA2 peptide can stimulate immune responses, suggesting its potential for targeted glioma vaccines.
Area of Science:
- Oncology
- Immunology
- Molecular Biology
Background:
- EphA2 receptor tyrosine kinase is overexpressed in advanced cancers.
- EphA2 expression is restricted in primary glioblastoma and anaplastic astrocytoma tissues compared to normal brain.
- Targeting EphA2 is a potential strategy for glioma vaccine development.
Purpose of the Study:
- To evaluate the potential of targeting EphA2 in glioma vaccine strategies.
- To assess antigen-specific cytotoxic T lymphocyte (CTL) responses against EphA2 in vitro and in vivo.
Main Methods:
- Stimulation of human leukocyte antigen (HLA) A2+ peripheral blood mononuclear cells (PBMCs) with EphA2883-891 peptide-loaded dendritic cells (DCs).
- Detection of antigen-specific CTL responses via lysis of peptide-loaded T2 cells and HLA-A2+ glioma cells (SNB19, U251).
- In vivo immunization of HLA-A2 transgenic HHD mice with the EphA2883-891 peptide.
Main Results:
- Stimulated PBMCs demonstrated antigen-specific CTL responses against EphA2.
- CTLs effectively lysed HLA-A2+ glioma cells expressing EphA2.
- In vivo immunization induced an epitope-specific CTL response in mice, even with an autoantigen.
Conclusions:
- The EphA2883-891 peptide elicits specific CTL responses.
- EphA2883-891 shows promise as a target antigen for molecularly targeted glioma vaccines.
