CTLs are targeted to kill β cells in patients with type 1 diabetes through recognition of a glucose-regulated preproinsulin epitope

Skowera, Ania; Ellis, Richard J.; Varela-Calviño, Rubén; Arif, S.A.M.; Guo, Cuicui; Van-Krinks, Cassie; Zaremba, Anna; Rackham, Chloe L.; Allen, Jeffrey; Tree, Timothy; Zhao, Minghui; Dayan, Colin; Sewell, Andrew K.; Unger, Wendy W. J.; Drijfhout, Jan W.; Ossendorp, Ferry; Roep, Bart O.; Peakman, Marie‐Claire

doi:10.1172/jci35449c1

Cited by 12 publications

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“…Immunoaffinity purification of HLA class I molecules and peptide extraction were carried out as previously described ( 5 ). Peptides were resuspended in 100/0.1 v/v water/formic acid and analyzed by an online nano high-performance liquid chromatography–mass spectrometry system ( 10 ).…”

Section: Methodsmentioning

confidence: 99%

“…HLA-A24 tetramers labeled with phycoerythrin (PE) and allophycocyanin (APC) were generated as described ( 5 ) and loaded with either PPI 3–11 (LWMRLLPLL) or the immunodominant, HLA-A24–presented cytomegalovirus (CMV) peptide (IE1 248–257 , AYAQKIFKIL; referred to as CMV-AYA) ( 11 , 12 ). Patient and control subject samples were examined for the presence of tetramer-positive CD8 T cells using 3 × 10 6 PBMCs washed twice with PBS and stained with a live/dead violet amine reactive dye (Invitrogen) for 15 min at room temperature, followed by washing and incubation with 100 nmol/L protein kinase inhibitor (dasatinib) ( 13 ) for 30 min at 37°C, 5% CO 2 .…”

Section: Methodsmentioning

confidence: 99%

“…CD8 T-cell clones specific for PPI 3–11 were generated as described ( 5 ). In brief, blood monocytes were cultured in X-Vivo 15 medium (Lonza, Walkersville, MD) supplemented with 5% AB, 100 IU/mL penicillin and 100 μg/mL streptomycin (Invitrogen), 100 ng/mL interleukin (IL)-4, and 50 ng/mL granulocyte-microphage colony-stimulating factor (Biosource/Invitrogen, Paisley, Scotland, U.K.) to obtain monocyte-derived immature dendritic cells (iDCs), which were harvested on day 6 and cryopreserved.…”

Section: Methodsmentioning

confidence: 99%

“…As described previously ( 5 ), islet-enriched cell fractions were cultured in CMRL1066 containing 5.6 mmol/L glucose, supplemented with 10% FCS, 100 units/mL penicillin, 100 μg/mL streptomycin, 2 mmol/L l -Gln (Invitrogen), 500 IU/mL interferon (IFN)-γ, 50 IU/mL IL-1β, 2,500 IU/mL TNF-α (all Miltenyi), and 1,000 IU/mL IFN-α (Roche, Mannheim, Germany) to upregulate HLA class I expression ( 5 ). Cytotoxicity was analyzed by a nonradioactive europium hydrophilic ligand cytotoxicity assay using DELFIA Technology (PerkinElmer) as described ( 5 ). When using K562 cell lines, 1 × 10 6 K562-A24 cells were pulsed with 10 µg PPI 3–11 for 1 h at 37°C or left unpulsed (K562-A24-PPI and K562-A24).…”

Section: Methodsmentioning

confidence: 99%

“…Several studies show an increased expression of HLA class I on islet cells in type 1 diabetic patients ( 1 , 3 , 4 ), which implies an enhanced susceptibility to CD8 T cell–mediated killing. Finally, in a previous study, we recapitulated the CD8 T-cell pathway of selective β-cell death in vitro by showing that T-cell clones generated from the blood of a patient, which recognize a peptide of preproinsulin (PPI) presented by HLA-A2*0201, can mediate specific β-cell killing ( 5 ).…”

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confidence: 99%

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Circulating Preproinsulin Signal Peptide–Specific CD8 T Cells Restricted by the Susceptibility Molecule HLA-A24 Are Expanded at Onset of Type 1 Diabetes and Kill β-Cells

et al. 2012

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Type 1 diabetes results from T cell–mediated β-cell destruction. The HLA-A*24 class I gene confers significant risk of disease and early onset. We tested the hypothesis that HLA-A24 molecules on islet cells present preproinsulin (PPI) peptide epitopes to CD8 cytotoxic T cells (CTLs). Surrogate β-cell lines secreting proinsulin and expressing HLA-A24 were generated and their peptide ligandome examined by mass spectrometry to discover naturally processed and HLA-A24–presented PPI epitopes. A novel PPI epitope was identified and used to generate HLA-A24 tetramers and examine the frequency of PPI-specific T cells in new-onset HLA-A*24+ patients and control subjects. We identified a novel naturally processed and HLA-A24–presented PPI signal peptide epitope (PPI3–11; LWMRLLPLL). HLA-A24 tetramer analysis reveals a significant expansion of PPI3–11-specific CD8 T cells in the blood of HLA-A*24+ recent-onset patients compared with HLA-matched control subjects. Moreover, a patient-derived PPI3–11-specific CD8 T-cell clone shows a proinflammatory phenotype and kills surrogate β-cells and human HLA-A*24+ islet cells in vitro. These results indicate that the type 1 diabetes susceptibility molecule HLA-A24 presents a naturally processed PPI signal peptide epitope. PPI-specific, HLA-A24–restricted CD8 T cells are expanded in patients with recent-onset disease. Human islet cells process and present PPI3–11, rendering themselves targets for CTL-mediated killing.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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Circulating Preproinsulin Signal Peptide–Specific CD8 T Cells Restricted by the Susceptibility Molecule HLA-A24 Are Expanded at Onset of Type 1 Diabetes and Kill β-Cells

et al. 2012

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Thymus-Derived Regulatory T Cells Are Positively Selected on Natural Self-Antigen through Cognate Interactions of High Functional Avidity

et al. 2016

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Regulatory T (Treg) cells expressing Foxp3 transcripton factor are essential for immune homeostasis. They arise in the thymus as a separate lineage from conventional CD4(+)Foxp3(-) T (Tconv) cells. Here, we show that the thymic development of Treg cells depends on the expression of their endogenous cognate self-antigen. The formation of these cells was impaired in mice lacking this self-antigen, while Tconv cell development was not negatively affected. Thymus-derived Treg cells were selected by self-antigens in a specific manner, while autoreactive Tconv cells were produced through degenerate recognition of distinct antigens. These distinct modes of development were associated with the expression of T cell receptor of higher functional avidity for self-antigen by Treg cells than Tconv cells, a difference subsequently essential for the control of autoimmunity. Our study documents how self-antigens define the repertoire of thymus-derived Treg cells to subsequently endow this cell type with the capacity to undermine autoimmune attack.

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The protection of Gα_z-null NOD mice from hyperglycemia is sexually dimorphic and only partially β-cell autonomous

et al. 2021

Preprint

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The mechanisms that underlie the β-cell pathophysiology of Type 1 Diabetes (T1D) are not fully understood. Our group has defined the unique heterotrimeric G protein alpha-subunit, Gαz, as a key negative regulator of β-cell signal transduction pathways. Non-obese diabetic (NOD) mice lacking Gαz throughout the body are protected from developing T1D-like hyperglycemia. To determine whether this phenotype is β-cell autonomous, we generated and validated a β-cell-specific Gαz knockout (βKO) on the NOD background and characterized the phenotype of female and male cohorts. Long-term hyperglycemia incidence was lower in Gαz βKO mice as compared to wild-type (WT) controls, but, unlike global Gαz knockout mice, this protection was incomplete. While young male and female Gαz βKO NOD mice had improved glucose tolerance, WT NOD males were significantly less glucose tolerant than females, and only female Gαz βKO mice retained improved glucose tolerance at 28-29 weeks of age. Conversely, β-cell-specific Gαz loss only influenced insulitis in 28-29-week old male NOD mice, a phenotype correlating directly with body burden of glucose during oral glucose challenge. Using surrogates for β-cell function and apoptosis, the partial penetrance of euglycemia in Gαz βKO NOD was best explained by an early failure to up-regulate β-cell proliferation. We conclude β-cell Gαz is an important regulator of the sexually-dimorphic T1D-like phenotype of NOD mice. Yet, other factors must be important in imparting full protection from the disease.

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CTLs are targeted to kill β cells in patients with type 1 diabetes through recognition of a glucose-regulated preproinsulin epitope

Abstract: CTLs are targeted to kill β cells in patients with type 1 diabetes through recognition of a glucose-regulated preproinsulin epitope

Cited by 12 publications

References 0 publications

Circulating Preproinsulin Signal Peptide–Specific CD8 T Cells Restricted by the Susceptibility Molecule HLA-A24 Are Expanded at Onset of Type 1 Diabetes and Kill β-Cells

Circulating Preproinsulin Signal Peptide–Specific CD8 T Cells Restricted by the Susceptibility Molecule HLA-A24 Are Expanded at Onset of Type 1 Diabetes and Kill β-Cells

Thymus-Derived Regulatory T Cells Are Positively Selected on Natural Self-Antigen through Cognate Interactions of High Functional Avidity

The protection of Gα_z-null NOD mice from hyperglycemia is sexually dimorphic and only partially β-cell autonomous

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CTLs are targeted to kill β cells in patients with type 1 diabetes through recognition of a glucose-regulated preproinsulin epitope

Abstract: CTLs are targeted to kill β cells in patients with type 1 diabetes through recognition of a glucose-regulated preproinsulin epitope

Cited by 12 publications

References 0 publications

Circulating Preproinsulin Signal Peptide–Specific CD8 T Cells Restricted by the Susceptibility Molecule HLA-A24 Are Expanded at Onset of Type 1 Diabetes and Kill β-Cells

Circulating Preproinsulin Signal Peptide–Specific CD8 T Cells Restricted by the Susceptibility Molecule HLA-A24 Are Expanded at Onset of Type 1 Diabetes and Kill β-Cells

Thymus-Derived Regulatory T Cells Are Positively Selected on Natural Self-Antigen through Cognate Interactions of High Functional Avidity

The protection of Gαz-null NOD mice from hyperglycemia is sexually dimorphic and only partially β-cell autonomous

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The protection of Gα_z-null NOD mice from hyperglycemia is sexually dimorphic and only partially β-cell autonomous