Journal: Experimental and Therapeutic Medicine

Article Title: Stability and inhibitory function of Treg cells under inflammatory conditions in vitro

doi: 10.3892/etm.2019.7873

Figure Lengend Snippet: Number of Treg cells following treatment with or without rhIL-6. Number of all Treg cells after culture for 3, 5, 7, 10 and 14 days in the presence of rhIL-2 (300 U/ml) and anti-CD3/CD28 beads (Treg cells/beads, 1:1) (n=8), without or with 25 ng/ml rhIL-6. Unpaired Student's t-test was used to compare differences between groups without and with rhIL-6. Values are expressed as the mean ± standard error of the mean (n=8). *P<0.05, Treg cells cultured for 7 days without vs. with rhIL-6 and **P<0.01, Treg cells cultured for 14 days without vs. with rhIL-6. rhIL, recombinant human interleukin; Treg cells, T-regulatory cells.

Article Snippet: Freshly sorted Treg cells were cultured (1×10 5 /well) in a 96-well U-bottomed plate in the presence of 100, 300 or 500 U/ml rhIL-2 (HumanZyme; ProteinTech Group, Inc.) and anti-CD3/CD28 beads (Treg cells/beads ratio, 0.5:1, 1:1 or 2:1), with or without 25 ng/ml rhIL-6 (HumanZyme) under 37°C in a humidified atmosphere containing 5% CO 2 .

Techniques: Cell Culture, Recombinant

Journal: Experimental and Therapeutic Medicine

Article Title: Stability and inhibitory function of Treg cells under inflammatory conditions in vitro

doi: 10.3892/etm.2019.7873

Figure Lengend Snippet: Proportion of CD4 + FoxP3 + Treg cells in all cell populations under inflammatory conditions. Percentages of CD4 + FoxP3 + Treg cells among (A) all cell populations and (B) the FoxP3 MFI of CD4 + FoxP3 + Treg cells after culture for 3, 5, 7, 10 and 14 days in the presence of rhIL-2 (300 U/ml) and anti-CD3/CD28 beads (Treg cells/beads, 1:1), with or without 25 ng/ml rhIL-6. Unpaired Student's t-test was used to compare differences between groups without and with rhIL-6. Values are expressed as the mean ± standard error of the mean (n=8). *P<0.05 vs. Treg cells cultured with rhIL-6. MFI, mean fluorescence intensity; Treg cells, T-regulatory cells; FoxP3, forkhead box P3; rhIL, recombinant human interleukin.

Article Snippet: Freshly sorted Treg cells were cultured (1×10 5 /well) in a 96-well U-bottomed plate in the presence of 100, 300 or 500 U/ml rhIL-2 (HumanZyme; ProteinTech Group, Inc.) and anti-CD3/CD28 beads (Treg cells/beads ratio, 0.5:1, 1:1 or 2:1), with or without 25 ng/ml rhIL-6 (HumanZyme) under 37°C in a humidified atmosphere containing 5% CO 2 .

Techniques: Cell Culture, Fluorescence, Recombinant

Journal: Experimental and Therapeutic Medicine

Article Title: Stability and inhibitory function of Treg cells under inflammatory conditions in vitro

doi: 10.3892/etm.2019.7873

Figure Lengend Snippet: Proportion of CD39 + CD4 + FoxP3 + active Treg cells in the CD4 + FoxP3 + cell populations. Percentages of CD39 + CD4 + FoxP3 + active Treg cells among (A) the CD4 + FoxP3 + Treg cells and (B) the CD39 MFI of CD39 + CD4 + FoxP3 + Treg cells after culture for 3, 5, 7, 10 and 14 days in the presence of rhIL-2 (300 U/ml) and anti-CD3/CD28 beads (Treg cells/beads, 1:1), with or without 25 ng/ml rhIL-6. Unpaired Student's t-test was used to compare differences between groups without and with rhIL-6. Values are expressed as the mean ± standard error of the mean (n=8). There was no statically significant differences were observed between the CD39 expression of the Treg cells between the two groups (P>0.05). MFI, mean fluorescence intensity; Treg cells, T-regulatory cells; FoxP3, forkhead box P3; rhIL, recombinant human interleukin.

Article Snippet: Freshly sorted Treg cells were cultured (1×10 5 /well) in a 96-well U-bottomed plate in the presence of 100, 300 or 500 U/ml rhIL-2 (HumanZyme; ProteinTech Group, Inc.) and anti-CD3/CD28 beads (Treg cells/beads ratio, 0.5:1, 1:1 or 2:1), with or without 25 ng/ml rhIL-6 (HumanZyme) under 37°C in a humidified atmosphere containing 5% CO 2 .

Techniques: Expressing, Fluorescence, Recombinant

Journal: Experimental and Therapeutic Medicine

Article Title: Stability and inhibitory function of Treg cells under inflammatory conditions in vitro

doi: 10.3892/etm.2019.7873

Figure Lengend Snippet: Suppression of Treg cells under inflammatory conditions. (A) Representative histograms indicating the CFSE dilution from a sample cultured without rhIL-6 and a sample cultured with 25 ng/ml rhIL-6. The proliferation curve of Teff cells cultured alone (red) was overlaid with the curve for Teff cells co-cultured with Treg cells (blue). (B-D) The sorted Teff cells from healthy blood donors (n=8) were labelled with CFSE and co-cultured with sorted Treg cells (Teff cells/Treg cells, 2:1) in the presence of anti-CD3/CD28 beads (Teff cells/beads, 1:1), without or with 25 ng/ml rhIL-6. After co-culture for 5 days, Teff-cell proliferation was assessed by flow cytometry, and (B) the proportion of responding T-cells, (C) doubling time of a single T-cell and (D) proliferative capacity for the Teff cells were calculated. Paired Student's t-test was used to compare differences between groups without and with rhIL-6. Values are expressed as the mean ± standard error of the mean (n=8). *P<0.05 and **P<0.01 compared the doubling time of a single T-cell and proliferative capacity of Teff cells cultured for 5 days without vs. with Treg cells. Treg cells, T-regulatory cells; Teff cells, T effector cells; CFSE, 5,6-carboxyfluorescein succinimidyl ester; rhIL, recombinant human interleukin.

Article Snippet: Freshly sorted Treg cells were cultured (1×10 5 /well) in a 96-well U-bottomed plate in the presence of 100, 300 or 500 U/ml rhIL-2 (HumanZyme; ProteinTech Group, Inc.) and anti-CD3/CD28 beads (Treg cells/beads ratio, 0.5:1, 1:1 or 2:1), with or without 25 ng/ml rhIL-6 (HumanZyme) under 37°C in a humidified atmosphere containing 5% CO 2 .

Techniques: Cell Culture, Co-Culture Assay, Flow Cytometry, Recombinant