Measuring T cell receptor and T cell gene expression diversity in antigen-responsive human CD4+ T cells.
T cells have diversity in TCR, epitope recognition, and cytokine production, and can be used for immune monitoring. Furthermore, clonal expansion of TCR families in disease may provide opportunities for TCR-directed therapies. We developed methodology for sequencing expressed genes of TCR alpha and beta chains from single cells and applied this to vaccine (tetanus-toxoid)-responsive CD4(+) T cells. TCR alpha and beta chains were both successfully sequenced in 1309 (43%) of 3038 CD4(+) T cells yielding 677 different receptors. TRAV and TRBV gene usage differed between tetanus-toxoid-responsive and non-responsive cells (p=0.004 and 0.0002), and there was extensive TCR diversity in tetanus-toxoid-responsive cells within individuals. Identical TCRs could be recovered in different samples from the same subject: TCRs identified after booster vaccination were frequent in pre-booster memory T cells (31% of pre-booster TCR), and also identified in pre-booster vaccination naïve cells (6.5%). No TCR was shared between subjects, but tetanus toxoid-responsive cells sharing one of their TCR chains were observed within and between subjects. Coupling single-cell gene expression profiling to TCR sequencing revealed examples of distinct cytokine profiles in cells bearing identical TCR. Novel molecular methodology demonstrates extensive diversity of Ag-responsive CD4(+) T cells within and between individuals.
DFG Research Center for Regenerative Therapies Dresden, Technische Universität Dresden, Germany; Paul Langerhans Institute Dresden, German Center for Diabetes Research (DZD). Electronic address: anne.eugster@crt-dresden.de.
Anne
A
Eugster
Annett
A
Lindner
Anne-Kristin
AK
Heninger
Carmen
C
Wilhelm
Sevina
S
Dietz
Mara
M
Catani
Anette-G
AG
Ziegler
Ezio
E
Bonifacio
D000328:Adult; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D004247:DNA / Q000235:genetics; D005260:Female; D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D007156:Immunologic Memory; D007223:Infant; D008297:Male; D008875:Middle Aged; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000235:genetics*; D017422:Sequence Analysis, DNA / Q000379:methods*; D059010:Single-Cell Analysis / Q000379:methods*; D016176:T-Lymphocyte Subsets / Q000276:immunology*; D013745:Tetanus Toxoid / Q000276:immunology*
D016693:Receptors, Antigen, T-Cell, alpha-beta; D013745:Tetanus Toxoid; D004247:DNA
Gene expression; Human; T cell receptor; T cells