Pesquisa imunológica
Acesso livre
ISSN: 1745-7580
ISSN: 1745-7580
Background The number of antigen receptors, immunoglobulins (IG) or antibodies and T cell receptors (TR) of the adap-tive immune response in vertebrates with jaws is almost unlimited (2.1012 per individual in humans). This huge diversity results from complex mechanisms in the synthesis of the variable (V) domains, that include DNA molecular rearrangements of the V, diversity (D) and joining (J) genes, N-diversity at the V-(D)-J junc-tions and, for IG, somatic hypermutations. The specificity of the V domains is conferred by the complemen-tarity determining regions (CDR) and more particularly the CDR3. IMGT®, the international ImMunoGeneT-ics information system®, has developed online tools that provide a detailed and accurate sequence analy-sis of the V domains (IMGT/V-QUEST) and CDR3 (IMGT/JunctionAnalysis), based on IMGT-ONTOLOGY. However online analyses are limited to 50 sequences per batch. The challenge was to provide identical high-quality analysis for the huge number of sequences obtained by Next Generation Sequencing (NGS) high throughput and deep sequencing. Results IMGT® has developed IMGT/HighV-QUEST that analyses up to 150,000 IG or TR V domain sequences per batch and performs statistical analysis on the results of up to 450,000 sequences. IMGT/HighV-QUEST provides users with: (i) a friendly web interface for submission and results retrieval, (ii) high-quality detailed results of IMGT/V-QUEST and IMGT/JunctionAnalysis, based on the IMGT-ONTOLOGY concepts and IMGT Scientific chart rules, (iii) a standardized frame for NGS statistical analysis, based on ‘Results catego-ry’ (‘1 copy’, ‘More than 1’, ‘single allele’, ‘several alleles (or genes)’, (iv) detailed standardized statistical analysis tables and histograms (e.g., V, D and J usage, CDR3-IMGT lengths). Conclusions IMGT/HighV-QUEST has been freely available for use for academics on the IMGT® Home page (http://www.imgt.org) since October 2010. More than 123 million sequences were submitted during its first year. IMGT/HighV-QUEST is a key component for establishing reliable repertoires of IG and TR V domains. The-se repertoires will contribute to the individual immunoprofiles in diverse immune situations and on different B and T cell populations. They will also contribute to characterize potential therapeutic antibodies from combinatorial libraries.