Abstract
We established several immortalized human T cell lines by cotransfecting primary lymphocytes derived from breast cancer patients with human c-myc and human c-Ha-ras oncogenes. A CD8 positive (CD8+) killer T cell line, FT-8, exhibitedin vitro specific cytotoxicity to a human breast cancer cell line, MCF-7. The FT-8 cells suppressed the growth of MCF-7 cells transplanted to athymic mice. The cytotoxic reaction was caused via T cell antigen receptor (TCR) on MCF-7 cells, because monoclonal antibodies against the TCR inhibited the cytotoxicity of FT-8 cells. The TCR(α) cDNA of FT-8 was cloned by using a PCR amplification technique and expressed by a cell-freein vitro translation system. The TCR(α) protein recognized a target antigen of 32 KDa on MCF-7 cells.
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Abbreviations
- FCS:
-
fetal calf serum
- FITC:
-
fluorescein isothiocynate
- MAb:
-
monoclonal antibody
- PE:
-
phycoerythrin
- TCR:
-
T cell recepter
- TCR(α):
-
α chain of TCR
- TPBS:
-
PBS containing Tween 20
References
Okubo M, Sato N, Wada Y, Takahashi S, Torimoto K, Takahashi N, Sato T, Okazaki M, Asaishi K, Kikuchi K Identification by monoclonal antibody of the tumor antigen of a human autologous breast cancer cell that is involved in cytotoxicity by a cytotoxic T-cell clone. Cancer Research 1989; 49: 3950–4.
Mikuras P, Gunhild LW, Prem S, Dean M, Robert CG Transformation of human umbilical cord blood T cells by human T-cell leukemia/lymphoma virus. Proc Natl Acad Sci USA 1983; 80: 5402–6
Taniguchi M, Miller JFAP Specific suppressor factors produced by hybridomas derived from the fusion of enriched suppressor T cells and a T lymphoma cell line. J Exp Med 1978; 148: 373–6.
Mavillio F, Sposi NM, Petrini M, Bottero L, Marinucci M, de Rossi G, Amadori S, Mandelli F, Peshele C Expression of cellular oncogenes in primary cells from human acute leukemia. Proc Natl Acad Sci USA 1986; 51: 5745–62.
Okano F, Tachibana H, Shirahata S, Murakami H Immortalization of human T lymphocytes by oncogenes. Cytotechnology 1993; in press.
Curran T, Macconnell WP, Van Straaten F, Verma IM Structure of the FBJ murine osteo sarcoma virus genome molecular cloning of its associated helper virus and the cellular homologue of the v-fos gene from mouse and human cells. Mol Cell Biol 1983; 3: 914–21.
Bergmann DG, Souza LM, Baluda MA Vertebrate DNA species contain nucleotide sequences related to the transforming gene of avian myeloblastosis virus. J Virol 1981; 40: 450–5.
Sekiya T, Fushimi M, Hori H, Hirohashi S, Nishimura S Molecular cloning and the total nucleotide sequence of the human c-Ha-ras-1 gene activated in a melanoma from a Japanese patient. Proc Natl Acad Sci USA 1983; 81: 4771–5.
Taya Y, Hosogai K, Hirohashi S, Shimosato Y, Tsuchiya R, Tsuchida N, Fushimi M, Sekiya T, Nishimura S A novel combination of K-ras andmyc amplification accompanied by point mutational activation of K-ras in a human lung cancer. EMBO J 1984; 3: 2943–6.
Land H, Parada LF, Weinberg RA Tumorigenic conversion of primary embryo fibroblasts requires at least two cooperating oncogenes. Nature 1983; 304: 596–602.
Tamai T, Shirahata S, Sato N, Nonaka, M, Murakami H. Purification and characterization of interferon-Iike antiviral protein derived from flatfish (Paralichthys olivaceus) lymphocytes immortalized by oncogenes. Cytotechnology, 11: 121–31.
Mosmann T Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assay. J Immunol Method 1983; 65: 55–63.
Aihara K, Yamada K, Murakami H, Nomura Y, Omura H Production of human-human hybridomas secreting monoclonal antibodies reactive to breast cancer cell line.In vitro Cellular & Developmental Biology 1988; 24: 959–62.
Ullrich A, Shine J, Chirgwin J, Pictet R, Tischer E, Rutter W Rat insulin gene: Construction of plasmids containing to coding sequences. Science 1977 169: 1313–9.
Gillis S Long term culture of tumor-specific cytotoxic T cells. Nature 1977; 268: 154–6.
Rosenberg S.A., Lotze M.T., Mull L.M. A progress report of the treatment of 157 patients with advanced cancer using lymphokine activated killer cells and interleukin 2 or high-dose of interleukin 2 alone. N Engl J Med 1987; 316: 889–94.
Rosenberg S.A., Spiess P, Lafreniere R A new approach to adoptive immunotherapy of cancer with tumor-infiltrating lymphocytes. Science 1986; 233: 1318–21.
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Okano, F., Tachibana, H., Akiyama, K. et al. The detection of a breast cancer antigen on MCF-7 cells reactive with the TCR(α) of a specific killer T cell line. Biotherapy 6, 195–203 (1993). https://doi.org/10.1007/BF01878081
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DOI: https://doi.org/10.1007/BF01878081