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A Syngeneic Mouse Model of Metastatic Renal Cell Carcinoma for Quantitative and Longitudinal Assessment of Preclinical Therapies
Published on: April 12, 2017
Establishment and characterization of a highly immunogenic human renal carcinoma cell line
Clelia Prattichizzo1, Margherita Gigante2, Paola Pontrelli2
1Department of Medical and Surgical Sciences, Section of Clinical Pathology, University of Foggia, Foggia, Italy.
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.
Insights
Researchers developed an immunogenic cell line, Elthem (RCC85#21), from clear cell carcinoma. This cell line effectively stimulates renal cell carcinoma (RCC)-specific T cells, offering a promising new avenue for RCC immunotherapy development.
Area of Science:
- Oncology
- Immunology
- Genetics
Background:
- Renal cell carcinoma (RCC) is the most common kidney cancer, often resistant to conventional treatments.
- Immunotherapy shows promise for RCC, with active specific immunotherapy and vaccine development being key targets.
- Identifying potent immunogenic antigens is crucial for boosting immune responses in RCC patients.
Purpose of the Study:
- To establish and characterize a highly immunogenic cell line for potential use in RCC immunotherapy.
- To assess the immunogenicity and T cell activation potential of the novel cell line, RCC85#21 (Elthem).
Main Methods:
- Genomic analysis (real-time PCR, genome instability) and proteomic analysis (2D electrophoresis, mass spectrometry) of the RCC85#21 cell line.
- Immunological assessment using mixed lymphocyte tumor cell cultures to evaluate T cell activation and cytotoxic responses.
- Analysis of T cell reactivity from RCC patients against the established cell line.
Main Results:
- The RCC85#21 cell line, derived from clear cell carcinoma, expressed tumor antigens and cytokine genes.
- Proteomic analysis identified 119 unique spots, while genomic analysis revealed loss of heterozygosity.
- RCC85#21 significantly activated RCC-specific T cells, demonstrating a cytotoxic phenotype and elevated reactivity with CD8+ T cells from RCC patients.
Conclusions:
- The established RCC85#21 immunogenic cell line is suitable for immune stimulation in renal cell carcinoma.
- The identification of novel tumor-associated antigens in RCC85#21 facilitates evaluation of immune responses.
- This work paves the way for developing novel immunotherapeutic strategies for renal cell carcinoma.
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