Characterization of Peripheral Blood TCR in Patients with Type 1 Diabetes Mellitus by BD RhapsodyTM VDJ CDR3 Assay

Takuro Okamura; Masahide Hamaguchi; Hiroyuki Tominaga; Noriyuki Kitagawa; Yoshitaka Hashimoto; Saori Majima; Takafumi Senmaru; Hiroshi Okada; Emi Ushigome; Naoko Nakanishi; Shigeyuki Shichino; Michiaki Fukui

doi:10.3390/cells11101623

Characterization of Peripheral Blood TCR in Patients with Type 1 Diabetes Mellitus by BD Rhapsody^TM VDJ CDR3 Assay

Cells. 2022 May 12;11(10):1623. doi: 10.3390/cells11101623.

Affiliations

¹ Department of Endocrinology and Metabolism, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto 602-8566, Japan.
² Division of Molecular Regulation of Inflammatory and Immune Diseases, Research Institute of Biomedical Sciences, Tokyo University of Science, Chiba 278-0022, Japan.

Abstract

The sequence of complementarity-determining region 3 of the T-cell receptor (TCR) varies widely due to the insertion of random bases during V-(D)-J recombination. In this study, we used single-cell VDJ sequencing using the latest technology, BD Rhapsody, to identify the TCR sequences of autoreactive T-cells characteristic of Japanese type 1 diabetes mellitus (T1DM) and to clarify the pairing of TCR of peripheral blood mononuclear cells from four patients with T1DM at the single-cell level. The expression levels of the TCR alpha variable (TRAV) 17 and TRAV21 in T1DM patients were higher than those in healthy Japanese subjects. Furthermore, the Shannon index of CD8⁺ T cells and FOXP3⁺ cells in T1DM patients was lower than that of healthy subjects. The gene expression of PRF1, GZMH, ITGB2, NKG7, CTSW, and CST7 was increased, while the expression of CD4, CD7, CD5, HLA-A, CD27, and IL-32 was decreased in the CD8⁺ T cells of T1DM patients. The upregulated gene expression was IL4R and TNFRSF4 in FOXP3⁺ cells of T1DM patients. Overall, these findings demonstrate that TCR diversity and gene expression of CD8⁺ and FOXP3⁺ cells are different in patients with T1DM and healthy subjects.

Keywords: T-cell receptor; genetic research; genetic screening; single-cell RNA sequencing; type 1 diabetes.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

CD8-Positive T-Lymphocytes* / metabolism
Diabetes Mellitus, Type 1* / metabolism
Forkhead Transcription Factors / metabolism
Humans
Leukocytes, Mononuclear
Receptors, Antigen, T-Cell / metabolism

Substances

Forkhead Transcription Factors
Receptors, Antigen, T-Cell

Associated data

figshare/10.6084/m9.figshare.19744570

Grants and funding

This research received funding from the Japan IDDM network, The Ichiro Kanehara Foundation for the Promotion of Medical Sciences and Medical Care, Foundation Kyoto Health Care Society, and the Uehara Memorial Foundation. This work was supported by KAKENHI, Grant-in-Aid for Young Scientists (20K17497). This work was supported by the Japan Agency for Medical Research and Development PRIME program (S.S., JP21gm6210025).