OPEN Research Support
head

Professor
Torben Barington
OPEN - Odense Patient data Explorative Network


Projekt styring
Projekt status    Sampling ongoing
 
Data indsamlingsdatoer
Start 01.01.2012  
Slut 01.01.2020  
 



Antibody repertoires and rearrangement mechanisms elucidated through next generation sequencing

Short summary

The purpose is to elucidate basic mechanisms involved in the generation of antibody specificity by sequencing large numbers of rearranged immunoglobulin genes (so called VDJ genes) and subjecting them to bioinformatics analyses.


Rationale

Human antibodies are formed by cells expressing a unique VDJ gene assembled by gene rearrangements in the bone marrow and later modified by somatic hypermutation in the lymph nodes. High throughput sequencing is a means to study the mechanisms behind these phenomena in detail. In this project, B cells from selected patients as well as from normal individuals are sequenced and bioinformatics tools developed allowing inference of basic mechanisms behind antibody production as well as pathogenic mechanisms involved in selected immunodeficiencies.  


Description of the cohort

B cells have been purified from blood samples from 120 healthy blood donors, anonymized and pooled. DNA was then extracted and subjected to PCR amplification and next generation sequencing of rearranged VDJ and DJ genes. These sequences were supplemented by publicly available VDJ sequences from the GeneBank database. 


Data and biological material

A total of 43,161 sequences including 34,100 VDH and 9,061 DJ sequences are available for analysis.

 


Collaborating researchers and departments

Department of Clinical Immunology, Odense University Hospital

  • Tina Funck
  • Mike Bogetofte Barnkob
  • Nanna Holm
  • Torben Barington
Department of Clinical Biochemistry, Zealand University Hospital, Roskilde

  • Tina Funck

 MRC Human Immunology Unit, Weatherall Institute of Molecular Medicine, University of Oxford, Oxfordshire

  • Mike Bogetofte Barnkob
Randall Division of Cell & Molecular Biophysics, Kings College London, London, UK

  • Line Ohm-Laursen
Department of ORL - Head & Neck Surgery, Odense University Hospital, Odense

  • Camilla Slot Mehlum

Publications associated with the project

Nucleotide Composition of Human Ig Nontemplated Regions Depends on Trimming of the Flanking Gene Segments, and Terminal Deoxynucleotidyl Transferase Favors Adding Cytosine, Not Guanosine, in Most VDJ Rearrangements. Funck T, Barnkob MB, Holm N, Ohm-Laursen L, Mehlum C, Möller S, Barington T. J Immunol. 2018 Sep 15;201(6):1765-1774. doi: 10.4049/jimmunol.1800100. Epub 2018 Aug 10.