title: High Throughput Sequencing of T Cell Antigen Receptors Reveals a Conserved TCR Repertoire.
abstract: The T-cell receptor (TCR) repertoire is a mirror of the human immune system that reflects processes caused by infections, cancer, autoimmunity, and aging. Next-generation sequencing has become a powerful tool for deep TCR profiling. Herein, we used this technology to study the repertoire features of TCR beta chain in the blood of healthy individuals.Peripheral blood samples were collected from 10 healthy donors. T cells were isolated with anti-human CD3 magnetic beads according to the manufacturer's protocol. We then combined multiplex-PCR, Illumina sequencing, and IMGT/High V-QUEST to analyze the characteristics and polymorphisms of the TCR.Most of the individual T cell clones were present at very low frequencies, suggesting that they had not undergone clonal expansion. The usage frequencies of the TCR beta variable, beta joining, and beta diversity gene segments were similar among T cells from different individuals. Notably, the usage frequency of individual nucleotides and amino acids within complementarity-determining region (CDR3) intervals was remarkably consistent between individuals. Moreover, our data show that terminal deoxynucleotidyl transferase activity was biased toward the insertion of G (31.92%) and C (27.14%) over A (21.82%) and T (19.12%) nucleotides.Some conserved features could be observed in the composition of CDR3, which may inform future studies of human TCR gene recombination.
authors.affiliation: From the State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou (XH, CL, QX, GC, JC, ZC, ZW, YD, PY, HD); Beijing Genomics Institute (SC); and Clinical Medical Research Center, The Second Clinical Medical College of Jinan University (Shenzhen People's Hospital) (YD), Shenzhen, Guangdong, China.
authors.forename: Xianliang
authors.initials: X
authors.lastname: Hou
authors.forename: Chong
authors.initials: C
authors.lastname: Lu
authors.forename: Sisi
authors.initials: S
authors.lastname: Chen
authors.forename: Qian
authors.initials: Q
authors.lastname: Xie
authors.forename: Guangying
authors.initials: G
authors.lastname: Cui
authors.forename: Jianing
authors.initials: J
authors.lastname: Chen
authors.forename: Zhi
authors.initials: Z
authors.lastname: Chen
authors.forename: Zhongwen
authors.initials: Z
authors.lastname: Wu
authors.forename: Yulong
authors.initials: Y
authors.lastname: Ding
authors.forename: Ping
authors.initials: P
authors.lastname: Ye
authors.forename: Yong
authors.initials: Y
authors.lastname: Dai
authors.forename: Hongyan
authors.initials: H
authors.lastname: Diao
mesh_terms: D000328:Adult; D022801:Complementarity Determining Regions / Q000235:genetics; D005260:Female; D014644:Genetic Variation; D064368:Healthy Volunteers; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods; D006801:Humans; D008297:Male; D008875:Middle Aged; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000235:genetics*; D013601:T-Lymphocytes / Q000502:physiology*
chemical_list: D022801:Complementarity Determining Regions; D016693:Receptors, Antigen, T-Cell, alpha-beta