Functional cloning of a gp100-reactive T-cell receptor from vitiligo patient skin

Jared Klarquist, Jonathan M. Eby, Steven W. Henning, Mingli Li, Derek A. Wainwright, Wiete Westerhof, Rosalie M. Luiten, Michael I. Nishimura, I. Caroline Le Poole*

*Corresponding author for this work

Research output: Contribution to journalArticle

6 Scopus citations

Abstract

We isolated gp100-reactive T cells from perilesional skin of a patient with progressive vitiligo with superior reactivity toward melanoma cells compared with tumor-infiltrating lymphocytes 1520, a melanoma-derived T-cell line reactive with the same cognate peptide. After dimer enrichment and limited dilution cloning, amplified cells were subjected to reverse transcription and 5' RACE to identify the variable TCRα and TCRβ subunit sequences. The full-length sequence was cloned into a retroviral vector separating both subunits by a P2A slippage sequence and introduced into Jurkat cells and primary T cells. Cytokine secreted by transduced cells in response to cognate peptide and gp100-expressing targets signifies that we have successfully cloned a gp100-reactive T-cell receptor from actively depigmenting skin.

Original languageEnglish (US)
Pages (from-to)379-384
Number of pages6
JournalPigment Cell and Melanoma Research
Volume29
Issue number3
DOIs
StatePublished - May 1 2016

Keywords

  • Melanoma
  • T cell receptor
  • Vitiligo

ASJC Scopus subject areas

  • Oncology
  • Biochemistry, Genetics and Molecular Biology(all)
  • Dermatology

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    Klarquist, J., Eby, J. M., Henning, S. W., Li, M., Wainwright, D. A., Westerhof, W., Luiten, R. M., Nishimura, M. I., & Le Poole, I. C. (2016). Functional cloning of a gp100-reactive T-cell receptor from vitiligo patient skin. Pigment Cell and Melanoma Research, 29(3), 379-384. https://doi.org/10.1111/pcmr.12458