Genome Technology Center

Immunorepertoire analysis by multiplex PCR and 454 sequencing

Jian Han, MD., Ph.D.
Faculty Investigator, HudsonAlpha Institute for Biotechnology

An immunorepertoire is the sum of functionally diverse B and T cells in one's circulation at any given moment. Three key factors shape the personal immunorepertoire: HLA type, antigen exposure history, and immunoregulation. Two major challenges have made studying the immunorepertoire difficult: the enormous diversity of V(D)Js, and the scarcity of each VDJ in a sample. To overcome these challenges, a powerful multiplex PCR method has been developed to amplify, inclusively and unbiased, all expressed VDJs in a sample. The amplicon mixture was sequenced directly with Roche 454 platform. In one run, for example, more than 73,000 and 119,000 unique V(D)Js were obtained from B and T cells, respectively. High resolution HLA typing was also carried out with the same sequencing run. This novel technology can be used to uncover possible disease mechanisms associated with abnormal immunorepertoires, to evaluate vaccine efficiency, to identify new biomarkers, and to develop new therapeutics. An online database and unique analytical tools have been developed for data sharing, deposition, and mining.

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