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Updated: Jun 17, 2025
High-Efficiency Generation of Antigen-Specific Primary Mouse Cytotoxic T Cells for Functional Testing in an Autoimmune Diabetes Model
Published on: August 16, 2019
Teplizumab induces persistent changes in the antigen-specific repertoire in individuals at risk for type 1 diabetes
Ana Lledó-Delgado1, Paula Preston-Hurlburt1, Sophia Currie1
1Departments of Immunobiology and Internal Medicine, Yale University School of Medicine, New Haven, Connecticut, USA.
Abstract:
BACKGROUNDTeplizumab, a non-FcR-binding anti-CD3 mAb, is approved to delay progression of type 1 diabetes (T1D) in at-risk patients. Previous investigations described the immediate effects of the 14-day treatment, but longer-term effects of the drug remain unknown.METHODSWith an extended analysis of study participants, we found that 36% were undiagnosed or remained free of clinical diabetes after 5 years, suggesting operational tolerance. Using single-cell RNA sequencing, we compared the phenotypes, transcriptome, and repertoire of peripheral blood CD8+ T cells including autoreactive T cells from study participants before and after teplizumab and features of responders and non-responders.RESULTSAt 3 months, there were transcriptional signatures of cell activation in CD4+ and CD8+ T cells including signaling that was reversed at 18 months. At that time, there was reduced expression of genes in T cell receptor and activation pathways in clinical responders. In CD8+ T cells, we found increased expression of genes associated with exhaustion and immune regulation with teplizumab treatment. These transcriptional features were further confirmed in an independent cohort. Pseudotime analysis showed differentiation of CD8+ exhausted and memory cells with teplizumab treatment. IL7R expression was reduced, and patients with lower expression of CD127 had longer diabetes-free intervals. In addition, the frequency of autoantigen-reactive CD8+ T cells, which expanded in the placebo group over 18 months, did not increase in the teplizumab group.CONCLUSIONThese findings indicate that teplizumab promotes operational tolerance in T1D, involving activation followed by exhaustion and regulation, and prevents expansion of autoreactive T cells.TRIAL REGISTRATIONClinicalTrials.gov NCT01030861.FUNDINGNational Institute of Diabetes and Digestive and Kidney Diseases/NIH, Juvenile Diabetes Research Foundation.
Insights
Teplizumab delays type 1 diabetes (T1D) progression by inducing operational tolerance, characterized by T cell activation, exhaustion, and regulation, preventing autoreactive T cell expansion.
Area of Science:
- Immunology
- Endocrinology
- Autoimmunity
Background:
- Teplizumab, an anti-CD3 mAb, is approved to delay type 1 diabetes (T1D) progression.
- Previous studies focused on immediate effects; longer-term impacts were unknown.
Purpose of the Study:
- To investigate the long-term effects of teplizumab on T cell phenotypes and function in T1D.
- To identify mechanisms of operational tolerance induced by teplizumab.
Main Methods:
- Single-cell RNA sequencing of CD8+ T cells from T1D patients before and after teplizumab treatment.
- Analysis of T cell receptor repertoire, transcriptome, and autoreactive T cell frequency.
- Comparison of responders and non-responders over 18 months.
Main Results:
- Teplizumab treatment led to transient T cell activation, followed by immune regulation and exhaustion signatures at 18 months.
- Clinical responders showed reduced T cell receptor and activation pathway gene expression.
- Autoreactive CD8+ T cell expansion was prevented in the teplizumab group, unlike the placebo group.
- Reduced IL7R (CD127) expression correlated with longer diabetes-free intervals.
Conclusions:
- Teplizumab promotes operational tolerance in T1D by modulating T cell responses.
- The drug induces a state of immune regulation and prevents the expansion of autoreactive T cells.
- Findings support teplizumab's role in managing T1D through immune modulation.
