Open Access

Establishment and characterization of a highly immunogenic human renal carcinoma cell line

  • Authors:
    • Clelia Prattichizzo
    • Margherita Gigante
    • Paola Pontrelli
    • Alessandro Stella
    • Maria Teresa Rocchetti
    • Maddalena Gigante
    • Eugenio Maiorano
    • Wolfgang Herr
    • Michele Battaglia
    • Loreto Gesualdo
    • Elena Ranieri
  • View Affiliations

  • Published online on: May 27, 2016     https://doi.org/10.3892/ijo.2016.3544
  • Pages: 457-470
  • Copyright: © Prattichizzo et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Renal cell carcinoma (RCC) is the most common kidney cancer, and accounts for ~3% of all adult malignancies. RCC has proven refractory to conventional treatment modalities but appears to be the only histological form that shows any consistent response to immunotherapeutic approaches. The development of a clinically effective vaccine remains a major strategic target for devising active specific immunotherapy in RCC. We aimed to identify a highly immunogenic antigenic format for immunotherapeutic approaches, so as to boost immune responses in RCC patients. We established and cloned an immunogenic cell line, RCC85#21 named Elthem, which was derived from a non-aggressive and non-metastatic clear cell carcinoma. The cell line characterization was performed by genomics (real-time PCR, genome instability), proteomics (two dimensional electrophoresis, mass spectro-metry) and immunological analysis (mixed lymphocytes tumor cell cultures). Real-time PCR confirmed the RCC85#21 cell expression of tumor antigens and cytokine genes. No difference in microsatellite instability (MSI) in RCC85#21 cell line was found as compared to control, loss of heterozygosity was observed in the RCC85#21 clone, but not in the renal cancer cell lines from which it was generated. The image analysis of RCC85#21 by two-dimensional gels showed 700±26 spots and 119 spots were identified by mass spectrometry analysis. RCC85#21 promoted a significant RCC-specific T cells activation by exhibiting a cytotoxic phenotype after mixed lymphocyte and tumor cell cultures. CD8+ T cells isolated from RCC patients displayed an elevated reactivity against RCC85#21 and efficiently lysed the RCC85#21 clone. The RCC85#21 immunogenic cell line will be suitable for immune stimulation. The identification of novel tumor associated antigens will allow the evaluation of the immune response in vitro and, subsequently, in vivo paving the way for new immunotherapeutic strategies in the RCC setting.
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August-2016
Volume 49 Issue 2

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

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Spandidos Publications style
Prattichizzo C, Gigante M, Pontrelli P, Stella A, Rocchetti MT, Gigante M, Maiorano E, Herr W, Battaglia M, Gesualdo L, Gesualdo L, et al: Establishment and characterization of a highly immunogenic human renal carcinoma cell line. Int J Oncol 49: 457-470, 2016
APA
Prattichizzo, C., Gigante, M., Pontrelli, P., Stella, A., Rocchetti, M.T., Gigante, M. ... Ranieri, E. (2016). Establishment and characterization of a highly immunogenic human renal carcinoma cell line. International Journal of Oncology, 49, 457-470. https://doi.org/10.3892/ijo.2016.3544
MLA
Prattichizzo, C., Gigante, M., Pontrelli, P., Stella, A., Rocchetti, M. T., Gigante, M., Maiorano, E., Herr, W., Battaglia, M., Gesualdo, L., Ranieri, E."Establishment and characterization of a highly immunogenic human renal carcinoma cell line". International Journal of Oncology 49.2 (2016): 457-470.
Chicago
Prattichizzo, C., Gigante, M., Pontrelli, P., Stella, A., Rocchetti, M. T., Gigante, M., Maiorano, E., Herr, W., Battaglia, M., Gesualdo, L., Ranieri, E."Establishment and characterization of a highly immunogenic human renal carcinoma cell line". International Journal of Oncology 49, no. 2 (2016): 457-470. https://doi.org/10.3892/ijo.2016.3544