High resolution analysis of the human t cell receptor repertoire

Krammer Manfred Schmidt and Christof von Kalle. High-throughput TCR sequencing TCR-seq involves the use of next generation sequencing platforms to generate large numbers of short DNA sequences covering key regions of the TCR coding sequence which enables quantification of T-cell diversity at unprecedented resolution.

Analysis of T-cell receptor TCR beta-chain gene expression in atherosclerotic lesions of human aorta.

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.

High-resolution analysis of the human T-cell receptor repertoire. Eliana Ruggiero 1 na1 Jan P. Nicolay 23 na1 Raffaele Fronza 1 Anne Arens 1 Anna Paruzynski 1 Ali Nowrouzi 1 Gökçe Ürenden 1.

High-resolution analysis of the human T-cell receptor repertoire. Eliana Ruggiero Department of Translational Oncology National Center for Tumor Diseases and German Cancer Research Center Im Neuenheimer Feld 280 69120 Heidelberg Germany. High-throughput T-cell receptor repertoire sequencing constitutes a powerful tool to study T cell responses at the clonal level.

However it does not give information on the functional phenotype. High-resolution analysis of the human T-cell receptor repertoire Eliana Ruggiero 1 Jan P. Nicolay 23 Raffaele Fronza 1 Anne Arens 1 Anna Paruzynski 1 Ali Nowrouzi 1.

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. 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-. High-resolution analysis of the human T-cell receptor repertoire By Eliana Ruggiero Jan P.

Nicolay Raffaele Fronza Anne Arens Anna Paruzynski Ali Nowrouzi Gökçe Ürenden Christina Lulay Sven Schneider Sergij Goerdt Hanno Glimm Peter H. Krammer Manfred Schmidt and Christof von Kalle. High-throughput analysis of the human thymic Vδ1 T cell receptor repertoire Download PDF.

High-throughput analysis of the human. High-throughput TCR sequencing TCR-seq involves the use of next generation sequencing platforms to generate large numbers of short DNA sequences covering key regions of the TCR coding sequence which enables quantification of T-cell diversity at unprecedented resolution. TCR-seq studies have provided new insights into the healthy human T-cell repertoire such as revised estimates of repertoire size and the understanding that TCR.

High-resolution analysis of the human T-cell receptor repertoire Nature Communications Sep 2015 Eliana Ruggiero Jan P. Nicolay Raffaele Fronza Anne Arens Anna Paruzynski Ali Nowrouzi Gökçe Ürenden Christina Lulay Sven Schneider Sergij Goerdt et al. The AbHelix assay is designed to sequence 5 chains targeting B cell receptors IgG IgM IgA IgK and IgL and 2 chains targeting T cell receptors TCRα and TCRβ.

The total RNA was divided evenly per B or T cell receptor chain type so only 27 of the total RNA provided was utilized for TCRα and TCRβ sequencing. The data from the B cell sequencing of HIP1 HIP2 and HIP3 at AbHelix was. Analysis of T-cell receptor TCR beta-chain gene expression in atherosclerotic lesions of human aorta.

TCR diversity was studied using non-radioactive polymerase chain reaction for quantitative assessment of TCRBV gene transcripts together with size and sequence analysis of the beta-chain third complementarity-determining region CDR3. Samples represent a wide range of atheromatous histology allowing evaluation of the T-cell repertoire. High-resolution analysis of the human T-cell receptor repertoire Published in.

Nature Communications September 2015 DOI. Eliana Ruggiero Jan P. Nicolay Raffaele Fronza Anne Arens Anna Paruzynski Ali Nowrouzi Gökçe Ürenden Christina Lulay Sven Schneider Sergij Goerdt Hanno Glimm Peter H.

High-resolution analysis of the human T-cell receptor repertoire. Eliana Ruggiero Jan P. Nicolay Raffaele Fronza Anne Arens Anna Paruzynski Ali Nowrouzi Gökçe Ürenden Christina Lulay Sven Schneider Sergij Goerdt Hanno Glimm Peter H.

Krammer Manfred Schmidt Christof Von Kalle. Contribution to journal Article. Recent advances in massively parallel sequencing have increased the depth at which T cell receptor TCR repertoires can be probed by 3log10 allowing for saturation sequencing of immune repertoires.

The resolution of this sequencing is dependent on its accuracy and direct assessments of the errors formed during high throughput repertoire analyses are limited. We analyzed 3 monoclonal TCR from TCR transgenic Rag– mice using Illumina sequencing. A total of 27 sequencing.

High-resolution analysis of the human T-cell receptor repertoire. Eliana Ruggiero Jan P. Nicolay Raffaele Fronza Anne Arens Anna Paruzynski Ali Nowrouzi Gökçe Ürenden Christina Lulay Sven Schneider Sergij Goerdt Hanno Glimm Peter H.

Krammer Manfred Schmidt Christof von Kalle. High-resolution analysis of human cytomegalovirus-specific T-cell receptor repertoires. The human herpesvirus cytomegalovirus CMV is a prevalent pathogen and infects a person for life.

After primary infection CMV latently resides in certain body cells to avoid clearance by the immune system. To gain insights into the ontogeny of the human innate B cell repertoire by performing high resolution analysis of the IgH-Cmu repertoire of human FL FBM and post-natal B cells. CD34negCD19pos B cells from human 2nd trimester FL and FBM child peripheral blood cPB and adult peripheral blood aPB were isolated by flow sorting.

For initial assessment of the IgM repertoire ontogeny in FL and FBM B cells we performed CDR3 fragment length analysis. High-resolution analysis of human cytomegalovirus-specific.