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

被引：69

作者：

Egorov, Evgeny S. ^{[1
]}

Merzlyak, Ekaterina M. ^{[1
]}

Shelenkov, Andrew A. ^{[2
]}

Britanova, Olga V. ^{[1
]}

Sharonov, George V. ^{[1
,3
]}

Staroverov, Dmitriy B. ^{[1
]}

Bolotin, Dmitriy A. ^{[1
]}

Davydov, Alexey N. ^{[4
]}

Barsova, Ekaterina ^{[1
]}

Lebedev, Yuriy B. ^{[1
]}

Shugay, Mikhail ^{[1
,4
,5
]}

Chudakov, Dmitriy M. ^{[1
,4
,5
]}

机构：

[1] Russian Acad Sci, Shemyakin Ovchinnikov Inst Bioorgan Chem, Moscow 117997, Russia

[2] Russian Acad Sci, Vavilov Inst Gen Genet, Moscow 119991, Russia

[3] Moscow MV Lomonosov State Univ, Fac Med, Moscow 119192, Russia

[4] Masaryk Univ, Cent European Inst Technol, CS-60177 Brno, Czech Republic

[5] Pirogov Russian Natl Res Med Univ, Moscow 117997, Russia

来源：

JOURNAL OF IMMUNOLOGY | 2015年 / 194卷 / 12期

基金：

俄罗斯科学基金会;

关键词：

T-CELL-RECEPTOR; ANTIBODY REPERTOIRE; DEEP; SEQUENCE; PCR; RECOMBINATION; LIBRARIES; RESPONSES; SELECTION; PLATFORM;

D O I：

10.4049/jimmunol.1500215

中图分类号：

R392 [医学免疫学]; Q939.91 [免疫学];

学科分类号：

071005 [微生物学]; 100108 [医学免疫学];

摘要：

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.

引用

页码：6155 / 6163

页数：9

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