Publication details

Quantitative Profiling of Immune Repertoires for Minor Lymphocyte Counts Using Unique Molecular Identifiers

Authors	EGOROV Evgeny S. MERZLYAK Ekaterina M. SHELENKOV Andrew A. BRITANOVA Olga V. SHARONOV George V. STAROVEROV Dmitriy B. BOLOTIN Dmitriy A. DAVYDOV Alexey Nikolayevich BARSOVA Ekaterina LEBEDEV Yuriy B. SHUGAY Mikhail CHUDAKOV Dmitriy
Year of publication	2015
Type	Article in Periodical
Magazine / Source	Journal of immunology
MU Faculty or unit	Central European Institute of Technology
Citation
Web	http://www.jimmunol.org/content/194/12/6155
Doi	http://dx.doi.org/10.4049/jimmunol.1500215
Field	Epidemiology, infectious diseases and clinical immunology
Keywords	T-CELL-RECEPTOR; ANTIBODY REPERTOIRE; DEEP; SEQUENCE; PCR; RECOMBINATION; LIBRARIES; RESPONSES; SELECTION; PLATFORM
Description	Emerging high-throughput sequencing methods for the analyses of complex structure of TCR and BCR repertoires give a powerful impulse to adaptive immunity studies. However, there are still essential technical obstacles for performing a truly quantitative analysis. Specifically, it remains challenging to obtain comprehensive information on the clonal composition of small lymphocyte populations, such as Ag-specific, functional, or tissue-resident cell subsets isolated by sorting, microdissection, or fine needle aspirates. In this study, we report a robust approach based on unique molecular identifiers that allows profiling Ag receptors for several hundred to thousand lymphocytes while preserving qualitative and quantitative information on clonal composition of the sample. We also describe several general features regarding the data analysis with unique molecular identifiers that are critical for accurate counting of starting molecules in high-throughput sequencing applications.