Journal: Arthritis & Rheumatology (Hoboken, N.j.)

Article Title: A CD8 T Cell/Indoleamine 2,3-Dioxygenase Axis Is Required for Mesenchymal Stem Cell Suppression of Human Systemic Lupus Erythematosus

doi: 10.1002/art.38674

Figure Lengend Snippet: Lupus CD8+ T cell–derived interferon-γ (IFNγ) promotes indoleamine 2,3-dioxygenase (IDO) production. A and B, Levels of IFNγ in supernatant are increased in the presence of UC-MSCs and SLE CD8+ T cells (A), while no change in levels of transforming growth factor β (TGFβ) is observed (B). Symbols represent individual subjects; horizontal lines show the mean (n = 7 per group). C, CD8+ T cells derived from patients with SLE produce much higher levels of intracellular IFNγ as compared to other cell subsets. D, Recombinant anti-human IFNγ antibody significantly abrogates SLE CD+ T cell–mediated IDO mRNA expression in UC-MSCs. E, Kynurenine levels in supernatant decrease in the presence of anti-human IFNγ antibody. F–H, In cocultured lupus peripheral blood mononuclear cells (PBMCs) and UC-MSCs, anti-human IFNγ antibodies (10 μg/ml) significantly inhibit kynurenine levels in supernatant (F), while the level of tryptophan is increased (G), and the ratio of kynurenine to tryptophan is decreased (H) (effects similar to those observed when 1-methyl- dl -tryptophan [1-DL-MT] is added). I, In the coculture system described above, UC-MSC–mediated inhibition of CD4+ T cell proliferation is abrogated by the addition of anti-human IFNγ antibody. Bars in C–I show the mean ± SEM (n = 7 per group in C, 8 per group in D, 8 per group in E, 5 per group in F–H, and 5 per group in I). ∗ = P < 0.05; ∗∗ = P < 0.01; ∗∗∗ = P < 0.001, by one-way ANOVA followed by Bonferroni test. PE = phycoerythrin; FITC = fluorescein isothiocyanate; APC = allophycocyanin (see for other definitions). Color figure can be viewed in the online issue, which is available at http://onlinelibrary.wiley.com/doi/10.1002/art.38674/abstract .

Article Snippet: The human TGFβ1 ELISA kit was from BioLegend.

Techniques: Derivative Assay, Recombinant, Expressing, Inhibition

Journal: Arthritis & Rheumatology (Hoboken, N.j.)

Article Title: A CD8 T Cell/Indoleamine 2,3-Dioxygenase Axis Is Required for Mesenchymal Stem Cell Suppression of Human Systemic Lupus Erythematosus

doi: 10.1002/art.38674

Figure Lengend Snippet: Defective indoleamine 2,3-dioxygenase (IDO) production in lupus bone marrow–derived MSCs (BM-MSCs). A–C, Expression of IFNGR1 (A), JAK-2 (B), and STAT-1 (C) was examined by real-time quantitative polymerase chain reaction. D, STAT-1, STAT-3, STAT-5, and their phosphorylated forms were assessed by Western blot analysis after treatment with UC-MSCs alone, UC-MSCs with different cell subsets (CD4+, CD8+, and non-CD4/CD8 T cells), or recombinant human interferon-γ (IFNγ). E, IDO mRNA expression in UC-MSCs, BM-MSCs from healthy controls, and BM-MSCs from SLE patients was examined after stimulation with different cytokines for 48 hours. F and G, Peripheral blood CD8+ T cells derived from patients with lupus were purified and used to stimulate UC-MSCs and BM-MSCs from healthy controls or lupus patients. Forty-eight hours later, IDO gene expression (F) and kynurenine enzyme activity (G) were determined. H, The ability of BM-MSCs from healthy controls and lupus patients to inhibit CD4+ T cell proliferation was compared. Bars in A–C and E–H show the mean ± SEM (n = 7 per group in A–D, 6 per group in E, 3 per group in F and G, and 4 per group in H). ∗∗ = P < 0.01; ∗∗∗ = P < 0.001, by one-way ANOVA. TGFβ = transforming growth factor β; IL-6 = interleukin-6; PBMC = peripheral blood mononuclear cell; NS = not significant (see for other definitions).

Article Snippet: The human TGFβ1 ELISA kit was from BioLegend.

Techniques: Derivative Assay, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Recombinant, Purification, Gene Expression, Activity Assay