ZnT8-Specific CD4+ T Cells Display Distinct Cytokine Expression Profiles between Type 1 Diabetes Patients and Healthy Adults

Daisuke Chujo, Emile Foucat, Thien Son Nguyen, Damien J. Chaussabel, Jacques Banchereau, Hideki Ueno

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Determination of antigen-specific T cell repertoires in human blood has been a challenge. Here, we show a novel integrated approach that permits determination of multiple parameters of antigen-specific T cell repertoires. The approach consists of two assays: the Direct assay and the Cytokine-driven assay. Briefly, human PBMCs are first stimulated with overlapping peptides encoding a given antigen for 48 hours to measure cytokine secretion (Direct assay). Peptide-reactive T cells are further expanded by IL-2 for 5 days; and after overnight starvation, expanded cells are stimulated with the same peptides from the initial culture to analyze cytokine secretion (Cytokine-driven assay). We first applied this integrated approach to determine the type of islet-antigen-specific T cells in healthy adults. Out of ten donors, the Direct assay identified GAD65-specific CD4+ T cells in three adults and zinc transporter 8 (ZnT8)-specific CD4+ T cells in five adults. The intracytoplasmic cytokine staining assay showed that these islet-antigen-specific CD4+ T cells belonged to the CD45RO+ memory compartment. The Cytokine-driven assay further revealed that islet-antigen-specific CD4+ T cells in healthy adults were capable of secreting various types of cytokines including type 1 and type 2 cytokines as well as IL-10. We next applied our integrated assay to determine whether the type of ZnT8-specific CD4+ T cells is different between Type 1 diabetes patients and age/gender/HLA-matched healthy adults. We found that ZnT8-specific CD4+ T cells were skewed towards Th1 cells in T1D patients, while Th2 and IL-10-producing cells were prevalent in healthy adults. In conclusion, the Direct assay and the Cytokine-driven assay complement each other, and the combination of the two assays provides information of antigen-specific T cell repertoires on the breadth, type, and avidity. This strategy is applicable to determine the differences in the quality of antigen-specific T cells between health and disease.

Original languageEnglish
Article numbere55595
JournalPLoS One
Volume8
Issue number2
DOIs
Publication statusPublished - 4 Feb 2013
Externally publishedYes

Fingerprint

T-cells
insulin-dependent diabetes mellitus
Medical problems
Type 1 Diabetes Mellitus
transporters
Assays
cytokines
T-lymphocytes
zinc
Display devices
Cytokines
T-Lymphocytes
assays
antigens
Antigens
CD4 Antigens
peptides
interleukin-10
Interleukin-10
Peptides

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

ZnT8-Specific CD4+ T Cells Display Distinct Cytokine Expression Profiles between Type 1 Diabetes Patients and Healthy Adults. / Chujo, Daisuke; Foucat, Emile; Nguyen, Thien Son; Chaussabel, Damien J.; Banchereau, Jacques; Ueno, Hideki.

In: PLoS One, Vol. 8, No. 2, e55595, 04.02.2013.

Research output: Contribution to journalArticle

Chujo, Daisuke ; Foucat, Emile ; Nguyen, Thien Son ; Chaussabel, Damien J. ; Banchereau, Jacques ; Ueno, Hideki. / ZnT8-Specific CD4+ T Cells Display Distinct Cytokine Expression Profiles between Type 1 Diabetes Patients and Healthy Adults. In: PLoS One. 2013 ; Vol. 8, No. 2.
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