T cell receptor reversed polarity recognition of a self-antigen major histocompatibility complex

Central to adaptive immunity is the interaction between the αβ T cell receptor (TCR) and peptide presented by the major histocompatibility complex (MHC) molecule. Presumably reflecting TCR-MHC bias and T cell signaling constraints, the TCR universally adopts a canonical polarity atop the MHC. We report the structures of two TCRs, derived from human induced T regulatory (iTreg) cells, complexed to an MHC class II molecule presenting a proinsulin-derived peptide. The ternary complexes revealed a 180° polarity reversal compared to all other TCR-peptide-MHC complex structures. Namely, the iTreg TCR α-chain and β-chain are overlaid with the α-chain and β-chain of MHC class II, respectively. Nevertheless, this TCR interaction elicited a peptide-reactive, MHC-restricted T cell signal. Thus TCRs are not 'hardwired' to interact with MHC molecules in a stereotypic manner to elicit a T cell signal, a finding that fundamentally challenges our understanding of TCR recognition

Infectious Tolerance as Candidate Therapy for Type 1 Diabetes: Transfer of Immunoregulatory Properties from Human Regulatory T Cells to Other T Cells and Proinflammatory Dendritic Cells

Transplantation and autoimmunit

Critical Role for TNF in the Induction of Human Antigen-Specific Regulatory T Cells by Tolerogenic Dendritic Cells

TNF is a pleiotropic cytokine with differential effects on immune cells and diseases. Anti-TNF therapy was shown to be effective in rheumatoid arthritis but proved inefficient or even detrimental in other autoimmune diseases. We studied the role of TNF in the induction of Ag-specific regulatory T cells (Tregs) by tolerogenic vitamin D3-modulated human dendritic cells (VD3-DCs), which previously were shown to release high amounts of soluble TNF (sTNF) upon maturation with LPS. First, production of TNF by modulated VD3-DCs was analyzed upon maturation with LPS or CD40L with respect to both secreted (cleaved) TNF (sTNF) and expression of the membrane-bound (uncleaved) form of TNF (mTNF). Next, TNF antagonists were tested for their effect on induction of Ag-specific Tregs by modulated DCs and the subsequent functionality of these Tregs. VD3-DCs expressed greater amounts of mTNF than did control DCs (nontreated DCs), independent of the maturation protocol. Inhibition of TNF with anti-TNF Ab (blocking both sTNF and mTNF) during the priming of Tregs with VD3-DCs prevented generation of Tregs and their suppression of proliferation of CD4(+) T cells. In contrast, sTNF receptor II (sTNFRII), mainly blocking sTNF, did not change the suppressive capacity of Tregs. Blocking of TNFRII by anti-CD120b Ab during Treg induction similarly abrogated their subsequent suppressive function. These data point to a specific role for mTNF on VD3-DCs in the induction of Ag-specific Tregs. Interaction between mTNF and TNFRII instructs the induction of suppressive Tregs by VD3-DCs. Anti-TNF therapy may therefore act adversely in different patients or disease pathways. The Journal of Immunology, 2010, 185: 1412-1418.Transplantation and autoimmunit

Differential Protein Pathways in 1,25-Dihydroxyvitamin D₃ and Dexamethasone Modulated Tolerogenic Human Dendritic Cells

Tolerogenic dendritic cells (DC) that are maturation-resistant and locked in a semimature state are promising tools in clinical applications for tolerance induction. Different immunomodulatory agents have been shown to induce a tolerogenic DC phenotype, such as the biologically active form of vitamin D (1,25(OH)₂D₃), glucocorticoids, and a synergistic combination of both. In this study, we aimed to characterize the protein profile, function and phenotype of DCs obtained <i>in vitro</i> in the presence of 1,25(OH)₂D₃, dexamethasone (DEX), and a combination of both compounds (combi). Human CD14⁺ monocytes were differentiated toward mature DCs, in the presence or absence of 1,25(OH)₂D₃ and/or DEX. Cells were prefractionated into cytoplasmic and microsomal fractions and protein samples were separated in two different pH ranges (pH 3–7NL and 6–9), analyzed by 2D-DIGE and differentially expressed spots (<i>p</i> < 0.05) were identified after MALDI-TOF/TOF analysis. In parallel, morphological and phenotypical analyses were performed, revealing that 1,25(OH)₂D₃- and combi-mDCs are closer related to each other than DEX-mDCs. This was translated in their protein profile, indicating that 1,25(OH)₂D₃ is more potent than DEX in inducing a tolerogenic profile on human DCs. Moreover, we demonstrate that combining 1,25(OH)₂D₃ with DEX induces a unique protein expression pattern with major imprinting of the 1,25(OH)₂D₃ effect. Finally, protein interaction networks and pathway analysis suggest that 1,25(OH)₂D₃, rather than DEX treatment, has a severe impact on metabolic pathways involving lipids, glucose, and oxidative phosphorylation, which may affect the production of or the response to ROS generation. These findings provide new insights on the molecular basis of DC tolerogenicity induced by 1,25(OH)₂D₃ and/or DEX, which may lead to the discovery of new pathways involved in DC immunomodulation

Differential Protein Pathways in 1,25-Dihydroxyvitamin D₃ and Dexamethasone Modulated Tolerogenic Human Dendritic Cells

Tolerogenic dendritic cells (DC) that are maturation-resistant and locked in a semimature state are promising tools in clinical applications for tolerance induction. Different immunomodulatory agents have been shown to induce a tolerogenic DC phenotype, such as the biologically active form of vitamin D (1,25(OH)₂D₃), glucocorticoids, and a synergistic combination of both. In this study, we aimed to characterize the protein profile, function and phenotype of DCs obtained <i>in vitro</i> in the presence of 1,25(OH)₂D₃, dexamethasone (DEX), and a combination of both compounds (combi). Human CD14⁺ monocytes were differentiated toward mature DCs, in the presence or absence of 1,25(OH)₂D₃ and/or DEX. Cells were prefractionated into cytoplasmic and microsomal fractions and protein samples were separated in two different pH ranges (pH 3–7NL and 6–9), analyzed by 2D-DIGE and differentially expressed spots (<i>p</i> < 0.05) were identified after MALDI-TOF/TOF analysis. In parallel, morphological and phenotypical analyses were performed, revealing that 1,25(OH)₂D₃- and combi-mDCs are closer related to each other than DEX-mDCs. This was translated in their protein profile, indicating that 1,25(OH)₂D₃ is more potent than DEX in inducing a tolerogenic profile on human DCs. Moreover, we demonstrate that combining 1,25(OH)₂D₃ with DEX induces a unique protein expression pattern with major imprinting of the 1,25(OH)₂D₃ effect. Finally, protein interaction networks and pathway analysis suggest that 1,25(OH)₂D₃, rather than DEX treatment, has a severe impact on metabolic pathways involving lipids, glucose, and oxidative phosphorylation, which may affect the production of or the response to ROS generation. These findings provide new insights on the molecular basis of DC tolerogenicity induced by 1,25(OH)₂D₃ and/or DEX, which may lead to the discovery of new pathways involved in DC immunomodulation

Differential Protein Pathways in 1,25-Dihydroxyvitamin D₃ and Dexamethasone Modulated Tolerogenic Human Dendritic Cells

Tolerogenic dendritic cells (DC) that are maturation-resistant and locked in a semimature state are promising tools in clinical applications for tolerance induction. Different immunomodulatory agents have been shown to induce a tolerogenic DC phenotype, such as the biologically active form of vitamin D (1,25(OH)₂D₃), glucocorticoids, and a synergistic combination of both. In this study, we aimed to characterize the protein profile, function and phenotype of DCs obtained <i>in vitro</i> in the presence of 1,25(OH)₂D₃, dexamethasone (DEX), and a combination of both compounds (combi). Human CD14⁺ monocytes were differentiated toward mature DCs, in the presence or absence of 1,25(OH)₂D₃ and/or DEX. Cells were prefractionated into cytoplasmic and microsomal fractions and protein samples were separated in two different pH ranges (pH 3–7NL and 6–9), analyzed by 2D-DIGE and differentially expressed spots (<i>p</i> < 0.05) were identified after MALDI-TOF/TOF analysis. In parallel, morphological and phenotypical analyses were performed, revealing that 1,25(OH)₂D₃- and combi-mDCs are closer related to each other than DEX-mDCs. This was translated in their protein profile, indicating that 1,25(OH)₂D₃ is more potent than DEX in inducing a tolerogenic profile on human DCs. Moreover, we demonstrate that combining 1,25(OH)₂D₃ with DEX induces a unique protein expression pattern with major imprinting of the 1,25(OH)₂D₃ effect. Finally, protein interaction networks and pathway analysis suggest that 1,25(OH)₂D₃, rather than DEX treatment, has a severe impact on metabolic pathways involving lipids, glucose, and oxidative phosphorylation, which may affect the production of or the response to ROS generation. These findings provide new insights on the molecular basis of DC tolerogenicity induced by 1,25(OH)₂D₃ and/or DEX, which may lead to the discovery of new pathways involved in DC immunomodulation

Differential Protein Pathways in 1,25-Dihydroxyvitamin D₃ and Dexamethasone Modulated Tolerogenic Human Dendritic Cells

Tolerogenic dendritic cells (DC) that are maturation-resistant and locked in a semimature state are promising tools in clinical applications for tolerance induction. Different immunomodulatory agents have been shown to induce a tolerogenic DC phenotype, such as the biologically active form of vitamin D (1,25(OH)₂D₃), glucocorticoids, and a synergistic combination of both. In this study, we aimed to characterize the protein profile, function and phenotype of DCs obtained <i>in vitro</i> in the presence of 1,25(OH)₂D₃, dexamethasone (DEX), and a combination of both compounds (combi). Human CD14⁺ monocytes were differentiated toward mature DCs, in the presence or absence of 1,25(OH)₂D₃ and/or DEX. Cells were prefractionated into cytoplasmic and microsomal fractions and protein samples were separated in two different pH ranges (pH 3–7NL and 6–9), analyzed by 2D-DIGE and differentially expressed spots (<i>p</i> < 0.05) were identified after MALDI-TOF/TOF analysis. In parallel, morphological and phenotypical analyses were performed, revealing that 1,25(OH)₂D₃- and combi-mDCs are closer related to each other than DEX-mDCs. This was translated in their protein profile, indicating that 1,25(OH)₂D₃ is more potent than DEX in inducing a tolerogenic profile on human DCs. Moreover, we demonstrate that combining 1,25(OH)₂D₃ with DEX induces a unique protein expression pattern with major imprinting of the 1,25(OH)₂D₃ effect. Finally, protein interaction networks and pathway analysis suggest that 1,25(OH)₂D₃, rather than DEX treatment, has a severe impact on metabolic pathways involving lipids, glucose, and oxidative phosphorylation, which may affect the production of or the response to ROS generation. These findings provide new insights on the molecular basis of DC tolerogenicity induced by 1,25(OH)₂D₃ and/or DEX, which may lead to the discovery of new pathways involved in DC immunomodulation

Differential Protein Pathways in 1,25-Dihydroxyvitamin D-3 and Dexamethasone Modulated Tolerogenic Human Dendritic Cells

Transplantation and autoimmunit