Novel approach to the characterization of melanoma associated-peptide-specific CTL lines from Japanese metastatic melanoma patients

  • Authors:
    • Yasuto Akiyama
    • Kouji Maruyama
    • Sachiko Tai
    • Masako Takikawa
    • Chie Ohshita
    • Akifumi Yamamoto
    • Naoya Yamazaki
    • Yoshio Kiyohara
    • Ken Yamaguchi
  • View Affiliations

  • Published online on: September 1, 2008     https://doi.org/10.3892/ijo_00000025
  • Pages: 433-441
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Abstract

Melanoma-associated antigens, MART-1, tyrosinase, gp100 and MAGEs, are typical melanoma-specific tumor antigens which can potently induce immune responses in metastatic melanoma patients treated with peptide vaccines. In the present study, we established a dendritic cell (DC)-based HLA-A2 melanoma-associated peptide (MART-1 or gp100)-specific CTL induction method and characterized the CTLs using HLA-A2 tetramer staining in 6 cases of HLA-A2+ melanoma treated with DC vaccines. Peripheral blood mononuclear cells (PBMC) from patients were stimulated twice with MART-1 A2 peptide-pulsed DCs in the presence of a low dose of IL-2. To boost CTL populations, CTL lines were further stimulated twice with MART-1 A2 peptide-pulsed T2 cells. The frequency of MART-1 A2 tetramer-positive CTLs increased from 0.16% (prior to stimulation) to 2.15% (after DC stimulation), and reached 46.5% on average (after additional T2 stimulation) in 4 cases which showed a successful expansion. The absolute numbers of MART-1 A2 tetramer-positive CTLs increased from 187- to 619-fold (average, 415-fold) compared to prior to DC stimulation. CTL assays using MART-1-specific CTL lines demonstrated potent killing activity against MART-1 peptide-pulsed T2 cells or HLA-A2+ melanoma cell lines in accordance with the frequency of tetramer-positive CTLs. Finally, we were successful in identifying melanoma peptide-specific T-cell receptor (TCR) cDNAs in 2 cases for MART-1 and 1 case for gp100 using the anti-TCR MoAb-based sorting as a novel approach instead of a conventional cell cloning, and confirmed peptide-specific IFN-γ production in TCR cDNA-transduced naïve T cells. The results showed that cloned TCR cDNAs were efficient in reconstituting tumor-specific cytotoxicity and good candidates for novel immunotherapy.

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September 2008
Volume 33 Issue 3

Print ISSN: 1019-6439
Online ISSN:1791-2423

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Spandidos Publications style
Akiyama Y, Maruyama K, Tai S, Takikawa M, Ohshita C, Yamamoto A, Yamazaki N, Kiyohara Y and Yamaguchi K: Novel approach to the characterization of melanoma associated-peptide-specific CTL lines from Japanese metastatic melanoma patients. Int J Oncol 33: 433-441, 2008.
APA
Akiyama, Y., Maruyama, K., Tai, S., Takikawa, M., Ohshita, C., Yamamoto, A. ... Yamaguchi, K. (2008). Novel approach to the characterization of melanoma associated-peptide-specific CTL lines from Japanese metastatic melanoma patients. International Journal of Oncology, 33, 433-441. https://doi.org/10.3892/ijo_00000025
MLA
Akiyama, Y., Maruyama, K., Tai, S., Takikawa, M., Ohshita, C., Yamamoto, A., Yamazaki, N., Kiyohara, Y., Yamaguchi, K."Novel approach to the characterization of melanoma associated-peptide-specific CTL lines from Japanese metastatic melanoma patients". International Journal of Oncology 33.3 (2008): 433-441.
Chicago
Akiyama, Y., Maruyama, K., Tai, S., Takikawa, M., Ohshita, C., Yamamoto, A., Yamazaki, N., Kiyohara, Y., Yamaguchi, K."Novel approach to the characterization of melanoma associated-peptide-specific CTL lines from Japanese metastatic melanoma patients". International Journal of Oncology 33, no. 3 (2008): 433-441. https://doi.org/10.3892/ijo_00000025