1. Immune Checkpoint Molecule, VISTA Regulates T-Cell-Mediated Skin Inflammatory Responses 
Tatsukuni Ohno, Yuta Kondo, Chenyang Zhang, Siwen Kang, Miyuki Azuma 
Journal of Investigative Dermatology 
Volume 137, Issue 6, Pages (June 2017)
DOI: /j.jid
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2. Figure 1
Treatment with anti-VISTA mAb at sensitization enhances CH responses. (a) DNFB was used for both sensitization and challenge. Either control Ig or anti-VISTA mAb was injected intraperitoneally 2 hours before each sensitization or challenge. Differences in ear thickness (ear swelling) before and 24, 48, and 72 hours after challenge were measured. The values are the mean ear thickness changes ± SD for each group of five mice. The data are representative of three independent experiments. (b) Histology (H&E) at 48 hours is shown. Scale bars = 25 μm. (c) IFN-γ and TNF-α secretion in challenged ear tissues are shown. The levels of IFN-γ and TNF-α in unsensitized and challenged mice were 6.8 ± 3.5 and 15.3 ± 10.2 pg/ml, respectively. The values are means ± SD for each group of four mice. The data shown are representative from two independent experiments. *P < (d) dLN cells from sensitized mice treated with control Ig or anti-VISTA mAb 4 days after the final sensitization were analyzed. Cells were stained with APC-anti-CD3 or -anti-Foxp3, FITC-anti-IFN-γ or anti-CD3, PerCp-Cy5.5-anti-CD4, and PE-anti-CD25 mAbs or with the appropriate fluorochrome-conjugated control Ig and analyzed by flow cytometry. Electronic gates were placed on CD4+CD3+ lymphocytes to identify CD4+ T cells and on CD4−CD3+ lymphocytes to identify CD8+ T cells, and the percentages of cells positive for CD25, IFN-γ, and Foxp3 were determined. All values are means ± SD for each group of three to five mice. The data shown are representative of two independent experiments. *P < APC, antigen-presenting cell; CH, contact hypersensitivity; dLN, draining lymph node; DNFB, 2,4-dinitro-1-fluorobenzene; H&E, hematoxylin and eosin; mAb, monoclonal antibody; SD, standard deviation; TNF-α, tumor necrosis factor-α; VISTA, V-domain Ig suppressor of T-cell activation.
Journal of Investigative Dermatology  , DOI: ( /j.jid )
Copyright © 2017 The Authors Terms and Conditions

3. Figure 2
Treatment with anti-VISTA mAb at sensitization modulates T-cell function. (a) T cells isolated from sensitized mice treated with control Ig or anti-VISTA mAb 4 days after the final sensitization were transferred into naïve mice. On the next day, DNFB challenge was performed and ear thickness was measured as described in Figure 1. The data shown are representative of two independent experiments. *P < (b) Nondepleted or CD4+ T-cell-depleted mice were subjected to sensitization and treatment with either control Ig or anti-VISTA mAb. After challenge, ear thickness changes were evaluated as described above. Values are means ± SD for each group of five mice. The data shown are representative of two independent experiments. (c) dLN-T cells (5 × 105 cells) from sensitized mice treated with control Ig or anti-VISTA mAb were cocultured with the indicated numbers of DNFB-pulsed dLN cells (left panel). dLN-T cells (5 × 105 cells) from sensitized mice were cocultured with the indicated numbers of either the control Ig- or anti-VISTA mAb-treated DNFB-pulsed APC fraction (right panel). Cultures were pulsed with [3H]-thymidine for the final 18 hours of the 72 hours’ culture. The values are means ± SD from triplicate cultures. *P < The data shown are representative of two independent experiments. APC, antigen-presenting cell; dLN, draining lymph node; DNFB, 2,4-dinitro-1-fluorobenzene; mAb, monoclonal antibody; SD, standard deviation; VISTA, V-domain Ig suppressor of T-cell activation.
Journal of Investigative Dermatology  , DOI: ( /j.jid )
Copyright © 2017 The Authors Terms and Conditions