Xinyu Xu1, Yong Gu1, Lingling Bian1, Yun Shi1, Yun Cai1, Yang Chen1, Heng Chen1, Li Qian1, Xiangmei Wu1, Kuanfeng Xu1, Roberto Mallone2, Howard W Davidson3, Liping Yu3, Jinxiong She4, Mei Zhang5, Tao Yang1. 1. Department of Endocrinology, First Affiliated Hospital of Nanjing Medical University, 300 Guangzhou Road, Nanjing, Jiangsu, China. 2. INSERM, U1016, Cochin Institute, Paris, France; CNRS, UMR8104, Cochin Institute, Paris, France; Paris Descartes University, Sorbonne Paris Cité, Paris, France; Assistance Publique Hôpitaux de Paris, Dept. of Diabetology, Cochin Hospital, Paris, France. 3. Department of Barbara Davis Center for Childhood Diabetes, University of Colorado at Denver and Health Sciences Center, USA. 4. Department of Endocrinology, First Affiliated Hospital of Nanjing Medical University, 300 Guangzhou Road, Nanjing, Jiangsu, China; Center for Biotechnology and Genomic Medicine, Medical College of Georgia, Georgia Regents University, Augusta, GA, USA. 5. Department of Endocrinology, First Affiliated Hospital of Nanjing Medical University, 300 Guangzhou Road, Nanjing, Jiangsu, China. Electronic address: zhangmei@njmu.edu.cn.
Abstract
OBJECTIVE: ZnT8-specific CD8+ T cells in human type 1 diabetes (T1D) have been reported recently, although the results from different laboratories are inconsistent. We aimed to characterize these ZnT8 specific CD8+ T cells and validate assays to screen peptide libraries. METHODS: We screened HLA-A2-restricted T cell candidate peptides of ZnT8 with different methods including computer algorithms, MHC-peptide binding and dissociation assays in T2 cell line, identification in HLA-A2 transgenic (Tg) mice and in vivo CTL assays. Then ELISpot assay was used to measure peptide-reactive T cell responses in 49 HLA-A2-restricted T1D patients. RESULTS: We demonstrated that ZnT8(107-116)(115), ZnT8(110-118), and ZnT8(177-186) were novel HLA-A*0201-restricted CTL epitopes in T1D patients. ZnT8(107-116)(115), ZnT8(115-123), ZnT8(153-161), ZnT8(177-186) and ZnT8(291-300) represent potentially major biomarkers for T1D. T cell responses against these epitopes showed different distributions between recently diagnosed and long-standing patients. Furthermore, they displayed discriminating performance among different ethnicities. We also compared the performance of the epitope identification strategies used herein. The epitopes which exhibited strong immunogenicity in HLA-A2 Tg mice were also well recognized by T1D patients. CONCLUSIONS: The differences in autoimmune T cell responses among T1D individuals may open new avenues toward T1D prediction and prevention. It also provides efficient strategies for immune intervention.
OBJECTIVE:ZnT8-specific CD8+ T cells in humantype 1 diabetes (T1D) have been reported recently, although the results from different laboratories are inconsistent. We aimed to characterize these ZnT8 specific CD8+ T cells and validate assays to screen peptide libraries. METHODS: We screened HLA-A2-restricted T cell candidate peptides of ZnT8 with different methods including computer algorithms, MHC-peptide binding and dissociation assays in T2 cell line, identification in HLA-A2 transgenic (Tg) mice and in vivo CTL assays. Then ELISpot assay was used to measure peptide-reactive T cell responses in 49 HLA-A2-restricted T1D patients. RESULTS: We demonstrated that ZnT8(107-116)(115), ZnT8(110-118), and ZnT8(177-186) were novel HLA-A*0201-restricted CTL epitopes in T1D patients. ZnT8(107-116)(115), ZnT8(115-123), ZnT8(153-161), ZnT8(177-186) and ZnT8(291-300) represent potentially major biomarkers for T1D. T cell responses against these epitopes showed different distributions between recently diagnosed and long-standing patients. Furthermore, they displayed discriminating performance among different ethnicities. We also compared the performance of the epitope identification strategies used herein. The epitopes which exhibited strong immunogenicity in HLA-A2 Tg mice were also well recognized by T1D patients. CONCLUSIONS: The differences in autoimmune T cell responses among T1D individuals may open new avenues toward T1D prediction and prevention. It also provides efficient strategies for immune intervention.
Authors: Simi Ahmed; Karen Cerosaletti; Eddie James; S Alice Long; Stuart Mannering; Cate Speake; Maki Nakayama; Timothy Tree; Bart O Roep; Kevan C Herold; Todd M Brusko Journal: Diabetes Date: 2019-07 Impact factor: 9.461
Authors: Joseph J Sabatino; Michael R Wilson; Peter A Calabresi; Stephen L Hauser; Jonathan P Schneck; Scott S Zamvil Journal: Proc Natl Acad Sci U S A Date: 2019-11-20 Impact factor: 11.205