Poster Presentation The 13th International Congress of the Immunology of Diabetes Society 2013

An Integrated Approach to Determine ZnT8-specific T Cell Repertoire in Type 1 Diabetes Patients and Healthy Adults (#72)

Daisuke Chujo 1 , Emile Foucat 2 , Thien-Son Nguyen 3 , Damien Chaussabel 3 , Masayuki Shimoda 1 , Shinichi Matsumoto 1 , Kunimasa Yagi 4 , Jacques Banchereau 2 , Hideki Ueno 2
  1. National Center for Global Health and Medicine, Tokyo, Japan
  2. Baylor Institute for Immunology Research, Dallas, TX, USA
  3. Benaroya Research Institute, Seattle, WA, USA
  4. Kanazawa University Graduate School of Medical Science, Kanazawa, Japan

Determination of antigen-specific T cell repertoires in human has been a challenge. Here, we show a novel integrated approach that permits determination of multiple parameters of antigen-specific T cell repertoires. Briefly, human PBMCs are first stimulated with overlapping peptides encoding a given antigen for 48 hours to measure cytokine production (Direct assay). Peptide-reactive T cells are further expanded by IL-2 for 5 days and re-stimulated with the same peptides from the initial culture to analyze cytokine production (Cytokine-driven assay). We first applied this approach to determine the type of ZnT8-specific T cells in healthy adults. Out of ten donors, the Direct assay identified ZnT8-specific CD4+ T cells in five adults. Furthermore, these ZnT8-specific CD4+ T cells belonged to the CD45RO+ memory compartment. The Cytokine-driven assay further revealed that those specific CD4+ T cells were capable of producing 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 (n=15) and age/gender/HLA-matched healthy subjects (n=15). We found that ZnT8-specific CD4+ T cells were skewed towards Th1 cells in T1D patients (p=0.0001 in the percentage of IL-10-IL-13-IFN-γ+CD4+ T cells), while Th2 and IL-10-producing cells were prevalent in healthy subjects (p=0.03 and p=0.04 in the percentage of IL-10+IL-13+IFN-γ-CD4+ and IL-10+IL-13-IFN-γ-CD4+ T cells, respectively). 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