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Identification of a novel cancer-testis antigen CRT2 frequently expressed in various cancers using representational
Emiko Hayashi1, Yuriko Matsuzaki, Go Hasegawa
1Division of Cellular Signaling, Institute for Advanced Medical Research, Keio University School of Medicine, Tokyo, Japan.
Purpose:
Cancer-testis antigens are promising targets for cancer immunotherapy. Identification of additional cancer-testis antigens with frequent expression in various cancers was attempted using representational differential analysis (RDA) and immunogenicity evaluation.
Experimental Design:
cDNAs preferentially expressed in testis were enriched using RDA by subtraction between testis and normal tissues. Thirty clones showing cancer-testis-like expression based on EST database analysis were evaluated by reverse transcription-PCR. A potential antigen, CRT2, was identified and its expression was analyzed with a newly generated anti-CRT2 antibody. The immunogenicity of CRT2 was examined based on reactivity with serum immunoglobulin G (IgG) from cancer patients, using Western blot and ELISA analysis, and on in vitro induction of tumor-reactive CTLs from HLA-A24 transgenic mice and human peripheral blood lymphocytes.
Results:
CRT2 was expressed in elongated spermatids of testis among normal tissues and in various cancer cell lines and tissues. The recombinant CRT2 protein was recognized by serum IgG from patients with various cancers in Western blot and ELISA analyses. A CRT2-derived peptide was identified as an HLA-A24-restricted T-cell epitope that induced tumor-reactive CTLs.
Conclusion:
CRT2 was identified as a new cancer-testis antigen expressed in elongated spermatids of testis and in cancer tissues (particularly melanoma) that is recognized by serum IgG from cancer patients. An HLA-A24-restricted T-cell epitope capable of inducing tumor-reactive CTLs was identified, suggesting that CRT2 may be useful for cancer diagnosis and immunotherapy.
Insights
A new cancer-testis antigen, CRT2, was identified and found in testis and various cancers. CRT2 shows promise for cancer immunotherapy due to its immunogenicity and ability to induce tumor-reactive T-cells.
Area of Science:
- Immunology
- Oncology
- Molecular Biology
Background:
- Cancer-testis antigens are crucial targets for cancer immunotherapy.
- Identifying novel cancer-testis antigens with broad cancer expression is essential.
Purpose of the Study:
- To identify new cancer-testis antigens using representational differential analysis (RDA).
- To evaluate the immunogenicity of identified antigens for potential therapeutic applications.
Main Methods:
- RDA was used to enrich testis-specific cDNAs.
- Reverse transcription-PCR and EST database analysis identified candidate clones.
- CRT2 expression and immunogenicity were assessed using antibody-based assays, Western blot, ELISA, and in vitro T-cell induction.
Main Results:
- CRT2 expression was confirmed in testis (elongated spermatids) and various cancer tissues/cell lines.
- Serum IgG from cancer patients recognized recombinant CRT2 protein.
- A CRT2-derived peptide induced tumor-reactive CTLs in an HLA-A24-restricted manner.
Conclusions:
- CRT2 is a novel cancer-testis antigen with expression in normal testis and diverse cancers.
- CRT2 elicits humoral and cellular immune responses in cancer patients.
- CRT2 and its epitopes hold potential for cancer diagnosis and immunotherapy.
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