Journal: Journal of the Endocrine Society

Article Title: Blood Lymphocyte Subsets and Proinflammatory Cytokine Profile in ROHHAD(NET) and non-ROHHAD(NET) Obese Individuals

doi: 10.1210/jendso/bvad103

Figure Lengend Snippet: Immunophenotype of PBMNC. The percentage of cells expressing different lineage markers for T cells (CD3, CD3/CD8, CD4, TCRαβ, TCRγδ), B cells (CD19), NK cells (CD16, CD56), regulatory B cells (Breg, CD19, CD38, and CD24), regulatory T cells (Treg, CD4, CD25, and CD127) and type 1 regulatory T cells (Tr1, CD4, CD45RO, LAG-3, and CD49b) has been evaluated by flow cytometry on PBMNC cells from patients affected by ROHHAD (n = 4, black squares), ROHHAD-NET (n = 4, gray triangles), and control individuals (n = 6, white dots). Results are expressed as number of cells/μL, which has been calculated as follows: (number of lymphocytes/μL × percentage of positive cells)/100. Horizontal bars indicated medians. P values are indicated where differences are statistically significant.

Article Snippet: Tr1 increase in our ROHHAD(NET) cohort could represent a consequence of the autoimmune activation in our patients, to be interpreted as an ineffective attempt to control such response, but this hypothesis needs to be confirmed.

Techniques: Expressing, Flow Cytometry, Control

Journal: Frontiers in Immunology

Article Title: Engineered T Regulatory Type 1 Cells for Clinical Application

doi: 10.3389/fimmu.2018.00233

Figure Lengend Snippet: T regulatory type 1 (Tr1)-mediated suppression in vivo . In steady-state condition, Tr1 cells reside in the spleen and circulate in the periphery. During inflammation, Tr1 cells are recruited to the site of tissue injury (i.e., after infections, autoimmune reactions, or transplantation) and are activated by professional antigen-presenting cells [APCs; dendritic cells (DCs)] via their T cell receptor, thus by their cognate antigen (Ag). Upon activation, Tr1 cells secrete IL-10 and TGF-β and (1) directly inhibit effector T cell (i.e., Th1 and Th17 cells) proliferation and pro-inflammatory cytokines production and (2) indirectly inhibit effector T cells by modulating professional APCs (i.e., downregulation of costimulatory and HLA class II expression and inhibition of pro-inflammatory cytokine secretion). (3) Tr1 cells can suppress effector T cells by cell-to-cell contact-mediated mechanisms, (4) suppress CD8 + T cell responses (i.e., proliferation and IFN-γ production), and (5) mediate bystander suppression by specifically killing professional APCs [DC or macrophages (M)], thus preventing naive T (Tn) cell priming and reactivation of effector T cells (i.e., Th1 and Th17 cells). Concomitantly, (6) Tr1 cells via IL-10 and TGF-β promote the induction of tolerogenic DC and anti-inflammatory macrophages (M2), which in turn promote de novo induction of Tr1 cells and T regulatory cells (Tregs), restoring tissue homeostasis and promoting long-term tolerance.

Article Snippet: An alternative method to induce/expand Ag-specific Tr1 cell medicinal product has been developed by the France-based company TxCell.

Techniques: In Vivo, Transplantation Assay, Activation Assay, Expressing, Inhibition

Journal: Frontiers in Immunology

Article Title: Engineered T Regulatory Type 1 Cells for Clinical Application

doi: 10.3389/fimmu.2018.00233

Figure Lengend Snippet: Protocols to generate/expand T regulatory type 1 (Tr1) cells. (A) Tr1-enriched cell lines. Donor-derived PBMC or CD4 + T cells are stimulated with host-derived monocytes in the presence of recombinant human IL-10 for 10 days. Alternatively, PBMC or CD4 + T cells are cultured for 10 days with allogenic dendritic cell (DC)-10, differentiated in vitro from peripheral blood monocytes with GM-CSF/IL-4/IL-10, in the presence of recombinant human IL-10 for 10 days . To generate T-allo10 cells, donor-derived T cells are cultured with host-derived DC-10 (Bacchetta and Roncarolo, unpublished data), whereas to induce host-derived T10 cells, T cells are stimulated with donor-derived DC-10 . (B) Tr1 cell clones. PBMC are stimulated with antigen (Ag; i.e., ovalbumin or collagen II) in the presence of IL-2 and IL-4 to enrich/expand Ag-specific T cells, followed by T cell cloning and expansion of the T cell clones using Schneider cells . (C) Tr1-like cell lines. Human CD4 + T cells are preactivated for 48 h with soluble anti-CD3/CD28 mAbs and IL-2 and then transduced with lentiviral vector (LV)-IL-10 overnight. Transduced T cells are isolated and expanded in feeder mixture . To generate allospecific IL-10-transduced cells, naive CD4 + T cells are stimulated with allogeneic DC and transduced with LV-IL-10 upon secondary stimulation. After selection, IL-10-trasduced cells are expanded in vitro with feeder mixture .

Article Snippet: An alternative method to induce/expand Ag-specific Tr1 cell medicinal product has been developed by the France-based company TxCell.

Techniques: Derivative Assay, Recombinant, Cell Culture, In Vitro, Clone Assay, Cloning, Transduction, Plasmid Preparation, Isolation, Selection

Journal: Frontiers in Immunology

Article Title: Engineered T Regulatory Type 1 Cells for Clinical Application

doi: 10.3389/fimmu.2018.00233

Figure Lengend Snippet: Tr1 cells in clinical and preclinical development.

Article Snippet: An alternative method to induce/expand Ag-specific Tr1 cell medicinal product has been developed by the France-based company TxCell.

Techniques: Clone Assay, In Vitro, In Vivo, Functional Assay