title: Functional cloning of a gp100-reactive T-cell receptor from vitiligo patient skin.
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.
authors.affiliation: Oncology Research Institute, Loyola University Chicago, Maywood, IL, USA.
authors.forename: Jared
authors.initials: J
authors.lastname: Klarquist
authors.affiliation: Oncology Research Institute, Loyola University Chicago, Maywood, IL, USA.
authors.forename: Jonathan M
authors.initials: JM
authors.lastname: Eby
authors.affiliation: Oncology Research Institute, Loyola University Chicago, Maywood, IL, USA.
authors.forename: Steven W
authors.initials: SW
authors.lastname: Henning
authors.affiliation: Department of Surgery, Medical University of South Carolina, Charleston, SC, USA.
authors.forename: Mingli
authors.initials: M
authors.lastname: Li
authors.affiliation: Oncology Research Institute, Loyola University Chicago, Maywood, IL, USA.
authors.forename: Derek A
authors.initials: DA
authors.lastname: Wainwright
authors.affiliation: Department of Dermatology, Netherlands Institute for Pigment Disorders, University of Amsterdam, AZ Amsterdam Zuidoost, The Netherlands.
authors.forename: Wiete
authors.initials: W
authors.lastname: Westerhof
authors.affiliation: Department of Dermatology, Netherlands Institute for Pigment Disorders, University of Amsterdam, AZ Amsterdam Zuidoost, The Netherlands.
authors.forename: Rosalie M
authors.initials: RM
authors.lastname: Luiten
authors.affiliation: Oncology Research Institute, Loyola University Chicago, Maywood, IL, USA.|Department of Surgery, Loyola University Chicago, Maywood, IL, USA.
authors.forename: Michael I
authors.initials: MI
authors.lastname: Nishimura
authors.affiliation: Oncology Research Institute, Loyola University Chicago, Maywood, IL, USA.|Departments of Pathology, Microbiology and Immunology, Loyola University Chicago, Maywood, IL, USA.
authors.forename: I Caroline
authors.initials: IC
authors.lastname: Le Poole
mesh_terms: D000328:Adult; D001483:Base Sequence; D002999:Clone Cells; D003001:Cloning, Molecular; D006801:Humans; D019169:Jurkat Cells; D011948:Receptors, Antigen, T-Cell / Q000378:metabolism*; D012867:Skin / Q000473:pathology*; D014820:Vitiligo / Q000235:genetics*; D058951:gp100 Melanoma Antigen / Q000235:genetics*
chemical_list: D011948:Receptors, Antigen, T-Cell; D058951:gp100 Melanoma Antigen
keywords: T cell receptor; melanoma; vitiligo
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