title: Peptide/MHC tetramer-based sorting of CD8⁺ T cells to a leukemia antigen yields clonotypes drawn nonspecifically from an underlying restricted repertoire.
abstract: Testing of T cell-based cancer therapeutics often involves measuring cancer antigen-specific T-cell populations with the assumption that they arise from in vivo clonal expansion. This analysis, using peptide/MHC tetramers, is often ambiguous. From a leukemia cell line, we identified a CDK4-derived peptide epitope, UNC-CDK4-1 (ALTPVVVTL), that bound HLA-A*02:01 with high affinity and could induce CD8⁺ T-cell responses in vitro. We identified UNC-CDK4-1/HLA-A*02:01 tetramer⁺ populations in 3 of 6 patients with acute myeloid leukemia who had undergone allogeneic stem cell transplantation. Using tetramer-based, single-cell sorting and T-cell receptor β (TCRβ) sequencing, we identified recurrent UNC-CDK4-1 tetramer-associated TCRβ clonotypes in a patient with a UNC-CDK4-1 tetramer⁺ population, suggesting in vivo T-cell expansion to UNC-CDK4-1. In parallel, we measured the patient's TCRβ repertoire and found it to be highly restricted/oligoclonal. The UNC-CDK4-1 tetramer-associated TCRβ clonotypes represented >17% of the entire TCRβ repertoire-far in excess of the UNC-CDK4-1 tetramer⁺ frequency-indicating that the recurrent TCRβ clonotypes identified from UNC-CDK-4-1 tetramer⁺ cells were likely a consequence of the extremely constrained T-cell repertoire in the patient and not in vivo UNC-CDK4-1-driven clonal T-cell expansion. Mapping recurrent TCRβ clonotype sequences onto TCRβ repertoires can help confirm or refute antigen-specific T-cell expansion in vivo.
authors.affiliation: Lineberger Comprehensive Cancer Center, Chapel Hill, North Carolina.
authors.forename: Sally A
authors.initials: SA
authors.lastname: Hunsucker
authors.affiliation: Lineberger Comprehensive Cancer Center, Chapel Hill, North Carolina.
authors.forename: Colleen S
authors.initials: CS
authors.lastname: McGary
authors.affiliation: Lineberger Comprehensive Cancer Center, Chapel Hill, North Carolina.
authors.forename: Benjamin G
authors.initials: BG
authors.lastname: Vincent
authors.affiliation: Department of Chemistry, University of North Carolina, Chapel Hill, North Carolina.
authors.forename: Atim A
authors.initials: AA
authors.lastname: Enyenihi
authors.affiliation: Department of Medicine, University of North Carolina, Chapel Hill, North Carolina.
authors.forename: Jennifer P
authors.initials: JP
authors.lastname: Waugh
authors.affiliation: Lineberger Comprehensive Cancer Center, Chapel Hill, North Carolina.
authors.forename: Karen P
authors.initials: KP
authors.lastname: McKinnon
authors.affiliation: Lineberger Comprehensive Cancer Center, Chapel Hill, North Carolina.
authors.forename: Lisa M
authors.initials: LM
authors.lastname: Bixby
authors.affiliation: Lineberger Comprehensive Cancer Center, Chapel Hill, North Carolina.
authors.forename: Patricia A
authors.initials: PA
authors.lastname: Ropp
authors.affiliation: Lineberger Comprehensive Cancer Center, Chapel Hill, North Carolina. Department of Medicine, University of North Carolina, Chapel Hill, North Carolina.
authors.forename: James M
authors.initials: JM
authors.lastname: Coghill
authors.affiliation: Lineberger Comprehensive Cancer Center, Chapel Hill, North Carolina. Department of Medicine, University of North Carolina, Chapel Hill, North Carolina.
authors.forename: William A
authors.initials: WA
authors.lastname: Wood
authors.affiliation: Lineberger Comprehensive Cancer Center, Chapel Hill, North Carolina. Department of Medicine, University of North Carolina, Chapel Hill, North Carolina.
authors.forename: Don A
authors.initials: DA
authors.lastname: Gabriel
authors.affiliation: Lineberger Comprehensive Cancer Center, Chapel Hill, North Carolina. Department of Medicine, University of North Carolina, Chapel Hill, North Carolina.
authors.forename: Stefanie
authors.initials: S
authors.lastname: Sarantopoulos
authors.affiliation: Lineberger Comprehensive Cancer Center, Chapel Hill, North Carolina. Department of Medicine, University of North Carolina, Chapel Hill, North Carolina.
authors.forename: Thomas C
authors.initials: TC
authors.lastname: Shea
authors.affiliation: Lineberger Comprehensive Cancer Center, Chapel Hill, North Carolina. Department of Medicine, University of North Carolina, Chapel Hill, North Carolina.
authors.forename: Jonathan S
authors.initials: JS
authors.lastname: Serody
authors.affiliation: Section of Transplantation Immunology, Department of Stem Cell Transplant and Cellular Therapy, The University of Texas MD Anderson Cancer Center, Houston, Texas.
authors.forename: Gheath
authors.initials: G
authors.lastname: Alatrash
authors.affiliation: Department of Melanoma, The University of Texas MD Anderson Cancer Center, Houston, Texas.
authors.forename: Tania
authors.initials: T
authors.lastname: Rodriguez-Cruz
authors.affiliation: Department of Melanoma, The University of Texas MD Anderson Cancer Center, Houston, Texas.
authors.forename: Gregory
authors.initials: G
authors.lastname: Lizée
authors.affiliation: Department of Immunobiology, University of Arizona, Tucson, Arizona.
authors.forename: Adam S
authors.initials: AS
authors.lastname: Buntzman
authors.affiliation: Department of Immunobiology, University of Arizona, Tucson, Arizona.
authors.forename: Jeffrey A
authors.initials: JA
authors.lastname: Frelinger
authors.affiliation: Department of Chemistry, University of North Carolina, Chapel Hill, North Carolina.
authors.forename: Gary L
authors.initials: GL
authors.lastname: Glish
authors.affiliation: Lineberger Comprehensive Cancer Center, Chapel Hill, North Carolina. Department of Medicine, University of North Carolina, Chapel Hill, North Carolina. paul_armistead@med.unc.edu.
authors.forename: Paul M
authors.initials: PM
authors.lastname: Armistead
mesh_terms: D000951:Antigens, Neoplasm / Q000276:immunology*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D018984:Epitopes, T-Lymphocyte / Q000276:immunology*; D005260:Female; D015789:HLA-A2 Antigen / Q000276:immunology; D006801:Humans; D007938:Leukemia / Q000276:immunology; D008297:Male; D008875:Middle Aged; D010455:Peptides / Q000276:immunology; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000276:immunology*; D016176:T-Lymphocyte Subsets / Q000276:immunology*; D020298:U937 Cells
chemical_list: D000951:Antigens, Neoplasm; D018984:Epitopes, T-Lymphocyte; C435939:HLA-A*02:01 antigen; D015789:HLA-A2 Antigen; D010455:Peptides; D016693:Receptors, Antigen, T-Cell, alpha-beta