High-resolution analysis of the human T-cell receptor repertoire

Unbiased dissection of T-cell receptor (TCR) repertoire diversity at the nucleotide level could provide important insights into human immunity. Here we show that TCR ligation-anchored-magnetically captured PCR (TCR-LA-MC PCR) identifies TCR α- and β-chain diversity without sequence-associated or quantitative restrictions in healthy and diseased conditions. TCR-LA-MC PCR identifies convergent recombination events, classifies different stages of cutaneous T-cell lymphoma in vivo and demonstrates TCR reactivation after in vitro cytomegalovirus stimulation. TCR-LA-MC PCR allows ultra-deep data access to both physiological TCR diversity and mechanisms influencing clonality in all clinical settings with restricted or distorted TCR repertoires.

[1]  S. Balk,et al.  Analysis of T cell antigen receptor (TCR) expression by human peripheral blood CD4-8- alpha/beta T cells demonstrates preferential use of several V beta genes and an invariant TCR alpha chain , 1993, The Journal of experimental medicine.

[2]  Steven J. M. Jones,et al.  Circos: an information aesthetic for comparative genomics. , 2009, Genome research.

[3]  Andrew K. Sewell,et al.  Genetic and Structural Basis for Selection of a Ubiquitous T Cell Receptor Deployed in Epstein-Barr Virus Infection , 2010, PLoS pathogens.

[4]  B. Pulendran,et al.  Ligation-anchored PCR: a simple amplification technique with single-sided specificity. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[5]  R. Küppers,et al.  Analysis of T-cell subpopulations in T-cell non-Hodgkin's lymphoma of angioimmunoblastic lymphadenopathy with dysproteinemia type by single target gene amplification of T cell receptor- beta gene rearrangements. , 2001, The American journal of pathology.

[6]  S. Migueles,et al.  Avidity for antigen shapes clonal dominance in CD8+ T cell populations specific for persistent DNA viruses , 2005, The Journal of experimental medicine.

[7]  J. Bell,et al.  Human HLA-A0201-restricted cytotoxic T lymphocyte recognition of influenza A is dominated by T cells bearing the V beta 17 gene segment , 1995, The Journal of experimental medicine.

[8]  M. Lieber,et al.  Mechanistic constraints on diversity in human V(D)J recombination , 1996, Molecular and cellular biology.

[9]  D. Douek,et al.  Unbiased Molecular Analysis of T Cell Receptor Expression Using Template‐Switch Anchored RT‐PCR , 2011, Current protocols in immunology.

[10]  Luca Biasco,et al.  Comprehensive genomic access to vector integration in clinical gene therapy , 2009, Nature Medicine.

[11]  Marie-Paule Lefranc,et al.  IMGT/JunctionAnalysis: the first tool for the analysis of the immunoglobulin and T cell receptor complex V-J and V-D-J JUNCTIONs , 2004, ISMB/ECCB.

[12]  Sang Joon Kim,et al.  A Mathematical Theory of Communication , 2006 .

[13]  W. Bossert,et al.  The Measurement of Diversity , 2001 .

[14]  Stephan Wolf,et al.  Genome-wide high-throughput integrome analyses by nrLAM-PCR and next-generation sequencing , 2010, Nature Protocols.

[15]  J. Bell,et al.  Variation in human T cell receptor V beta and J beta repertoire: analysis using anchor polymerase chain reaction. , 1992, European journal of immunology.

[16]  S. Josefowicz,et al.  CMV antigen-specific CD4+ and CD8+ T cell IFNgamma expression and proliferation responses in healthy CMV-seropositive individuals. , 2004, Viral immunology.

[17]  Daniel C. Douek,et al.  Convergent recombination shapes the clonotypic landscape of the naïve T-cell repertoire , 2010, Proceedings of the National Academy of Sciences.

[18]  J. Bell,et al.  Extensive conservation of alpha and beta chains of the human T-cell antigen receptor recognizing HLA-A2 and influenza A matrix peptide. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[19]  D J Moss,et al.  Dominant selection of an invariant T cell antigen receptor in response to persistent infection by Epstein-Barr virus , 1994, The Journal of experimental medicine.

[20]  F. Marincola,et al.  CMV pp65 and IE-1 T cell epitopes recognized by healthy subjects , 2007, Journal of Translational Medicine.

[21]  David A Jones,et al.  Profound loss of T-cell receptor repertoire complexity in cutaneous T-cell lymphoma. , 2003, Blood.

[22]  R. Holt,et al.  Profiling the T-cell receptor beta-chain repertoire by massively parallel sequencing. , 2009, Genome research.

[23]  Benedict Ng,et al.  NKT and MAIT invariant TCRα sequences can be produced efficiently by VJ gene recombination. , 2013, Immunobiology.

[24]  Mikhail Shugay,et al.  MiTCR: software for T-cell receptor sequencing data analysis , 2013, Nature Methods.

[25]  W. J. Kent,et al.  BLAT--the BLAST-like alignment tool. , 2002, Genome research.

[26]  Nicola Pimpinelli,et al.  Revisions to the staging and classification of mycosis fungoides and Sezary syndrome: a proposal of the International Society for Cutaneous Lymphomas (ISCL) and the cutaneous lymphoma task force of the European Organization of Research and Treatment of Cancer (EORTC). , 2007, Blood.

[27]  Jérôme Lane,et al.  IMGT®, the international ImMunoGeneTics information system® , 2004, Nucleic Acids Res..

[28]  Sergey Lukyanov,et al.  Next generation sequencing for TCR repertoire profiling: Platform‐specific features and correction algorithms , 2012, European journal of immunology.

[29]  C. Carlson,et al.  Overlap and Effective Size of the Human CD8+ T Cell Receptor Repertoire , 2010, Science Translational Medicine.

[30]  R. Küppers,et al.  Analysis of T-cell subpopulations in T-cell non-Hodgkin's lymphoma of angioimmunoblastic lymphadenopathy with dysproteinemia type by single target gene amplification of T cell receptor- beta gene rearrangements. , 2001, The American journal of pathology.

[31]  C E Shannon,et al.  The mathematical theory of communication. 1963. , 1997, M.D. computing : computers in medical practice.

[32]  M S Waterman,et al.  Identification of common molecular subsequences. , 1981, Journal of molecular biology.

[33]  Daniel C. Douek,et al.  A Novel Approach to the Analysis of Specificity, Clonality, and Frequency of HIV-Specific T Cell Responses Reveals a Potential Mechanism for Control of Viral Escape1 , 2002, The Journal of Immunology.

[34]  L. Picker,et al.  Cytomegalovirus (CMV) phosphoprotein 65 makes a large contribution to shaping the T cell repertoire in CMV-exposed individuals. , 2002, The Journal of infectious diseases.

[35]  Hanno Glimm,et al.  High-resolution insertion-site analysis by linear amplification–mediated PCR (LAM-PCR) , 2007, Nature Methods.

[36]  D. Price,et al.  The molecular basis for public T-cell responses? , 2008, Nature Reviews Immunology.

[37]  Abigail Wacher,et al.  Comprehensive assessment of T-cell receptor beta-chain diversity in alphabeta T cells. , 2009, Blood.

[38]  G. Crooks,et al.  WebLogo: a sequence logo generator. , 2004, Genome research.

[39]  Richard A. Moore,et al.  Exhaustive T-cell repertoire sequencing of human peripheral blood samples reveals signatures of antigen selection and a directly measured repertoire size of at least 1 million clonotypes. , 2011, Genome research.

[40]  Daniel C. Douek,et al.  Sharing of T cell receptors in antigen-specific responses is driven by convergent recombination , 2006, Proceedings of the National Academy of Sciences.

[41]  Jiahuai Han,et al.  Determinants of public T cell responses , 2012, Cell Research.

[42]  Frank Baas,et al.  Human T-cell memory consists mainly of unexpanded clones. , 2010, Immunology letters.

[43]  Baback Gharizadeh,et al.  High throughput sequencing reveals a complex pattern of dynamic interrelationships among human T cell subsets , 2010, Proceedings of the National Academy of Sciences.

[44]  O. Gotoh An improved algorithm for matching biological sequences. , 1982, Journal of molecular biology.

[45]  Susan M. Schlenner,et al.  Thymus-autonomous T cell development in the absence of progenitor import , 2012, The Journal of experimental medicine.

[46]  X. Jin,et al.  The human cytotoxic T-lymphocyte (CTL) response to cytomegalovirus is dominated by structural protein pp65: frequency, specificity, and T-cell receptor usage of pp65-specific CTL , 1996, Journal of virology.

[47]  M. Bonneville,et al.  Selection of T Cell Clones Expressing High-Affinity Public TCRs within Human Cytomegalovirus-Specific CD8 T Cell Responses1 , 2005, The Journal of Immunology.