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Liang Deng, Wanhong Ding, Richard D. Granstein 

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Presentation on theme: "Liang Deng, Wanhong Ding, Richard D. Granstein "— Presentation transcript:

1 Thalidomide Inhibits Tumor Necrosis Factor-α Production and Antigen Presentation by Langerhans Cells 
Liang Deng, Wanhong Ding, Richard D. Granstein  Journal of Investigative Dermatology  Volume 121, Issue 5, Pages (November 2003) DOI: /j x Copyright © 2003 The Society for Investigative Dermatology, Inc Terms and Conditions

2 Figure 1 Effect of thalidomide on TNF-α production in LPS-stimulated XS106A cells. A total of 5×106 XS106A cells were treated with thalidomide at various concentrations (50, 100, or 200 μg/mL) or DMSO before stimulation with LPS. Supernatants were collected at different time points (3, 9, and 21 h). Data represent TNF-α concentration in the supernatants, expressed as averages±SD from triplicates in one experiment. Statistical analysis shows the difference in the amount TNF-α production is significant at all concentration levels and at all time points tested, p<0.001 compared to no thalidomide. Journal of Investigative Dermatology  , DOI: ( /j x) Copyright © 2003 The Society for Investigative Dermatology, Inc Terms and Conditions

3 Figure 2 Inhibition of TNF-α production by thalidomide in LPS-stimulated XS52 cells. A total of 5×105 XS52 cells were treated with thalidomide at various concentrations (50, 100, or 200 μg/mL) or DMSO. Supernatants were taken out at 9 h after LPS stimulation. Data represent the concentration of TNF-α in the supernatants, expressed as averages±SD from triplicates in one experiment. *p<0.01 for no thalidomide versus all other groups. Journal of Investigative Dermatology  , DOI: ( /j x) Copyright © 2003 The Society for Investigative Dermatology, Inc Terms and Conditions

4 Figure 3 Inhibition of TNF-α production and increase of IL-12 production by thalidomide in LPS-stimulated XS52 cells at low concentrations. A total of 3×105 cells were treated with thalidomide at various concentrations (10, 20, or 50 μg/mL) or DMSO for 12 h. Supernatants were taken out at 9 h after LPS stimulation. Data represent the concentration of TNF-α, IL-12 p40 subunit in the supernatants, expressed as averages±SD from triplicates in one experiment. *p<0.05 for no thalidomide vs. thalidomide. Journal of Investigative Dermatology  , DOI: ( /j x) Copyright © 2003 The Society for Investigative Dermatology, Inc Terms and Conditions

5 Figure 4 Inhibition of TNF-α production by thalidomide in purified LC. Purified LC (4 × 105 cells/well) were treated with thalidomide at 200 μg per mL or DMSO as control for 24 h. Supernatants were taken out at 12 and 21 h after LPS administration. Data represent TNF-α concentration in the supernatants, expressed as averages±SD from triplicates in one experiment. *p<0.01 for both 12 and 21 h. Journal of Investigative Dermatology  , DOI: ( /j x) Copyright © 2003 The Society for Investigative Dermatology, Inc Terms and Conditions

6 Figure 5 The effect of thalidomide on TNF-α mRNA level in LPS-stimulated XS106A cells as analyzed by northern blot hybridization. Total RNA (25 μg) from XS106A cells treated with thalidomide (200 μg/mL) for 48 h and stimulated with LPS for 1, 3, 6, and 9 h was probed for TNF-α and β-actin. Autographs were exposed for 4 h for TNF-α and 2 h for β-actin. Journal of Investigative Dermatology  , DOI: ( /j x) Copyright © 2003 The Society for Investigative Dermatology, Inc Terms and Conditions

7 Figure 6 The effect of thalidomide on the immunostimulatory function of epidermal LC as determined by DO11.1 assay. Epidermal cells (EC) at different concentrations (0, 0.5 × 105, 1×105, and 2×105) were pretreated with thalidomide (200 μg/mL) or DMSO for 24 h. Then they were incubated with cOVA for 24 h. Cells were washed and then coincubated with DO11.1 cells for 24 h. Supernatants were collected and incubated with CTLL-2 cells. The incorporation of [3H]thymidine by CTLL-2 cells was measured by scintillation counter. Results represent averages±SD of triplicates. *p<0.001. Journal of Investigative Dermatology  , DOI: ( /j x) Copyright © 2003 The Society for Investigative Dermatology, Inc Terms and Conditions

8 Figure 7 The effects of thalidomide on the immunostimulatory function of epidermal LC as determined by HDK assay. Enriched LLs were preincubated with KLH, thalidomide at various concentrations (10, 25, 50, 100, or 200 μg/mL), or DMSO as control for 24 h. After they were washed, 1×105 ECs were coincubated with 5 × 104 HDK cells for 48 h. The amount of IFN-γ in the supernatants was measured by ELISA assay. HDK cells alone and LCs alone were used as negative controls. Results represent averages±SD of triplicates. p<0.05 for 0 vs. 10 μg per mL; p<0.001 for 0 vs. all other thalidomide groups. Journal of Investigative Dermatology  , DOI: ( /j x) Copyright © 2003 The Society for Investigative Dermatology, Inc Terms and Conditions

9 Figure 8 Thalidomide does not affect HDK cells directly. Fifty thousand HDK cells were treated with thalidomide directly at 10, 25, 50, 100, or 200 μg per mL overnight. They were then added to 1×105 EC cells that were preincubated with KLH for 24 h. Incubation was for 18 h. Supernatants were collected to evaluate IFN-γ production using ELISA assay. Results represent averages±SD of triplicates. Journal of Investigative Dermatology  , DOI: ( /j x) Copyright © 2003 The Society for Investigative Dermatology, Inc Terms and Conditions

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