title: Deep sequencing of the TCR-β repertoire of human forkhead box protein 3 (FoxP3)+and FoxP3-T cells suggests that they are completely distinct and non-overlapping.
abstract: Maintenance of peripheral tolerance requires a balance between autoreactive conventional T cells (Tconv) and thymically derived forkhead box protein 3 (FoxP3)+regulatory T cells (tTregs). Considerable controversy exists regarding the similarities/differences in T cell receptor (TCR) repertoires expressed by Tconvand tTregs. We generated highly purified populations of human adult and cord blood Tconvand tTregsbased on the differential expression of CD25 and CD127. The purity of the sorted populations was validated by intracellular staining for FoxP3 and Helios. We also purified an overlap group of CD4 T cells from adult donors to ensure that considerable numbers of shared clonotypes could be detected when present. We used deep sequencing of entire TCR-β CDR3 sequences to analyse the TCR repertoire of Tconvand tTregs. Our studies suggest that both neonatal and adult human Tconvand tTregcells are, in fact, entirely distinct CD4 T cell lineages.
authors.affiliation: Laboratory of Immunology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA.
authors.forename: A
authors.identifier: 0000-0003-3659-6654
authors.initials: A
authors.lastname: Golding
authors.affiliation: Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA.
authors.forename: S
authors.initials: S
authors.lastname: Darko
authors.affiliation: Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA.
authors.forename: W H
authors.initials: WH
authors.lastname: Wylie
authors.affiliation: Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA.
authors.forename: D C
authors.initials: DC
authors.lastname: Douek
authors.affiliation: Laboratory of Immunology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA.
authors.forename: E M
authors.initials: EM
authors.lastname: Shevach
mesh_terms: D015415:Biomarkers; D060965:Clonal Evolution; D022801:Complementarity Determining Regions / Q000235:genetics; D051858:Forkhead Transcription Factors / Q000378:metabolism*; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D016130:Immunophenotyping; D010641:Phenotype; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000235:genetics* / Q000378:metabolism; D016176:T-Lymphocyte Subsets / Q000276:immunology / Q000378:metabolism*; D050378:T-Lymphocytes, Regulatory / Q000276:immunology / Q000378:metabolism; D050441:VDJ Exons / Q000235:genetics
chemical_list: D015415:Biomarkers; D022801:Complementarity Determining Regions; C418974:FOXP3 protein, human; D051858:Forkhead Transcription Factors; D016693:Receptors, Antigen, T-Cell, alpha-beta
keywords: T cells repertoire; deep sequencing; regulatory T cells