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An HLA-restricted, p53 specific immune response from HLA transgenic p53 knockout mice

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Abstract

Background: p53 is over-expressed in most human malignancies and is therefore an attractive target for immunotherapy. Unfortunately, a human cytotoxic T cell response to p53 is difficult to generate. p53 knockout transgenic mice may provide a model to circumvent immunologic tolerance to p53 and develop high-affinity p53-specific cytotoxic T lymphocytes (CTL).

Methods: p53 knockout, HLA A2.1 transgenic mice were generated and immunized with the immunodominant wild-type p53 nonamer peptide epitope p53149-157. Two weeks later splenocytes were harvested and stimulated in vitro with acid-treated, p53 peptide-pulsed syngeneic blast cells. Cultures were restimulated weekly with acid-treated, p53 peptide-pulsed Jurkat cells transfected with the HLA A2.1 gene. Peptide-specific cytotoxic activity was measured by chromium release assay, and the resulting CD8+ effectors were cloned via limiting dilution.

Results: P53 peptide-specific CTL were generated against p53149-157. Clones generated from the p53149-157 cell line demonstrated high affinity and specificity for p53149-157 when presented by HLA A2.1+ antigen-presenting cells. The p53149-157 CTL killed only cells overexpressing p53 cells that were HLA A2.1+ and did not kill cells with normal levels of p53 expression or those that were HLA A2.1-.

Conclusion: HLA transgenic mice not previously exposed to the p53 protein provide a useful model for generating high-affinity p53-specific CTL.

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Authors and Affiliations

  1. Dept. of Surgery, City of Hope National Medical Center, 1500 E. Duarte Rd., 91010, Duarte, CA, USA

    Todd M. McCarty MD, Zhiwei Yu MD &  Joshua D. I. Ellenhorn MD

  2. the Department of Hematology, City of Hope National Medical Center, Duarte, California, USA

    Xiping Liu MD, PhD & Don J. Diamond PhD

Authors
  1. Todd M. McCarty MD
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  2. Zhiwei Yu MD
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  3. Xiping Liu MD, PhD
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  4. Don J. Diamond PhD
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  5. Joshua D. I. Ellenhorn MD
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McCarty, T.M., Yu, Z., Liu, X. et al. An HLA-restricted, p53 specific immune response from HLA transgenic p53 knockout mice. Annals of Surgical Oncology 5, 93–99 (1998). https://doi.org/10.1007/BF02303770

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  • DOI: https://doi.org/10.1007/BF02303770

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