Single-Cell RNA Sequencing Reveals Expanded Clones of Islet Antigen-Reactive CD4+ T Cells in Peripheral Blood of Subjects with Type 1 Diabetes

被引:64
作者
Cerosaletti, Karen [1 ]
Barahmand-pour-Whitman, Fariba [2 ]
Yang, Junbao [1 ]
DeBerg, Hannah A. [2 ]
Dufort, Matthew J. [2 ]
Murray, Sara A. [2 ]
Israelsson, Elisabeth [2 ]
Speake, Cate [3 ]
Gersuk, Vivian H. [2 ]
Eddy, James A. [2 ]
Reijonen, Helena [3 ]
Greenbaum, Carla J. [3 ]
Kwok, William W. [1 ]
Wambre, Erik [1 ]
Prlic, Martin [4 ]
Gottardo, Raphael [4 ]
Nepom, Gerald T. [5 ]
Linsley, Peter S. [2 ]
机构
[1] Virginia Mason, Benaroya Res Inst, Translat Res Program, 1201 Ninth Ave, Seattle, WA 98101 USA
[2] Virginia Mason, Syst Immunol Program, Benaroya Res Inst, 1201 Ninth Ave, Seattle, WA 98101 USA
[3] Virginia Mason, Benaroya Res Inst, Diabet Clin Res Program, Seattle, WA 98101 USA
[4] Fred Hutchinson Canc Res Ctr, Vaccine & Infect Dis Div, Seattle, WA 98109 USA
[5] Immune Tolerance Network, Bethesda, MD 20814 USA
基金
美国国家卫生研究院;
关键词
SUBUNIT-RELATED PROTEIN; RECEPTOR; IDENTIFICATION; AUTOANTIGEN; RECOGNITION; REPERTOIRE; PHENOTYPE; PEPTIDE; INSULIN; EPITOPE;
D O I
10.4049/jimmunol.1700172
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
The significance of islet Ag-reactive T cells found in peripheral blood of type 1 diabetes (T1D) subjects is unclear, partly because similar cells are also found in healthy control (HC) subjects. We hypothesized that key disease-associated cells would show evidence of prior Ag exposure, inferred from expanded TCR clonotypes, and essential phenotypic properties in their transcriptomes. To test this, we developed single-cell RNA sequencing procedures for identifying TCR clonotypes and transcript phenotypes in individual T cells. We applied these procedures to analysis of islet Ag-reactive CD4(+) memory T cells from the blood of T1D and HC individuals after activation with pooled immunodominant islet peptides. We found extensive TCR clonotype sharing in Ag-activated cells, especially from individual T1D subjects, consistent with in vivo T cell expansion during disease progression. The expanded clonotype from one T1D subject was detected at repeat visits spanning >15 mo, demonstrating clonotype stability. Notably, we found no clonotype sharing between subjects, indicating a predominance of "private" TCR specificities. Expanded clones from two T1D subjects recognized distinct IGRP peptides, implicating this molecule as a trigger for CD4(+) T cell expansion. Although overall transcript profiles of cells from HC and T1D subjects were similar, profiles from the most expanded clones were distinctive. Our findings demonstrate that islet Ag-reactive CD4(+) memory T cells with unique Ag specificities and phenotypes are expanded during disease progression and can be detected by single-cell analysis of peripheral blood.
引用
收藏
页码:323 / 335
页数:13
相关论文
共 67 条
[1]   A disease-associated cellular immune response in type 1 diabetics to an immunodominant epitope of insulin [J].
Alleva, DG ;
Crowe, PD ;
Jin, LP ;
Kwok, WW ;
Ling, N ;
Gottschalk, M ;
Conlon, PJ ;
Gottlieb, PA ;
Putnam, AL ;
Gaur, A .
JOURNAL OF CLINICAL INVESTIGATION, 2001, 107 (02) :173-180
[2]   Autoreactive T cell responses show proinflammatory polarization in diabetes but a regulatory phenotype in health [J].
Arif, S ;
Tree, TI ;
Astill, TP ;
Tremble, JM ;
Bishop, AJ ;
Dayan, CM ;
Roep, BO ;
Peakman, M .
JOURNAL OF CLINICAL INVESTIGATION, 2004, 113 (03) :451-463
[3]  
Auf der Maur P., 1979, HUM GENET, V49, P209
[4]   Antigen-Reactive T Cell Enrichment for Direct, High-Resolution Analysis of the Human Naive and Memory Th Cell Repertoire [J].
Bacher, Petra ;
Schink, Christian ;
Teutschbein, Janka ;
Kniemeyer, Olaf ;
Assenmacher, Mario ;
Brakhage, Axel A. ;
Scheffold, Alexander .
JOURNAL OF IMMUNOLOGY, 2013, 190 (08) :3967-3976
[5]   Single cell sequencing approaches for complex biological systems [J].
Baslan, Timour ;
Hicks, James .
CURRENT OPINION IN GENETICS & DEVELOPMENT, 2014, 26 :59-65
[6]   IMGT/V-QUEST: the highly customized and integrated system for IG and TR standardized V-J and V-D-J sequence analysis [J].
Brochet, Xavier ;
Lefranc, Marie-Paule ;
Giudicelli, Veronique .
NUCLEIC ACIDS RESEARCH, 2008, 36 :W503-W508
[7]  
Charles A, 2001, Immunobiology: The Immune System in Health and Disease, V5th
[8]   Assessment of CD4+T Cell Responses to Glutamic Acid Decarboxylase 65 Using DQ8 Tetramers Reveals a Pathogenic Role of GAD65 121-140 and GAD65 250-266 in T1D Development [J].
Chow, I-Ting ;
Yang, Junbao ;
Gates, Theresa J. ;
James, Eddie A. ;
Mai, Duy T. ;
Greenbaum, Carla ;
Kwok, William W. .
PLOS ONE, 2014, 9 (11)
[9]   Adult-onset type 1 diabetes patients display decreased IGRP-specific Tr1 cells in blood [J].
Chujo, Daisuke ;
Nguyen, Thien-Son ;
Foucat, Emile ;
Blankenship, Derek ;
Banchereau, Jacques ;
Nepom, Gerald T. ;
Chaussabel, Damien ;
Ueno, Hideki .
CLINICAL IMMUNOLOGY, 2015, 161 (02) :270-277
[10]   Hotspot autoimmune T cell receptor binding underlies pathogen and insulin peptide cross-reactivity [J].
Cole, David K. ;
Bulek, Anna M. ;
Dolton, Garry ;
Schauenberg, Andrea J. ;
Szomolay, Barbara ;
Rittase, William ;
Trimby, Andrew ;
Jothikumar, Prithiviraj ;
Fuller, Anna ;
Skowera, Ania ;
Rossjohn, Jamie ;
Zhu, Cheng ;
Miles, John J. ;
Peakman, Mark ;
Wooldridge, Linda ;
Rizkallah, Pierre J. ;
Sewell, Andrew K. .
JOURNAL OF CLINICAL INVESTIGATION, 2016, 126 (06) :2191-2204