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UVA-Induced Immune Suppression Through an Oxidative Pathway

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1 UVA-Induced Immune Suppression Through an Oxidative Pathway
Ichiro Iwai, Masato Hatao, Masako Naganuma, Yoshimaru Kumano  Journal of Investigative Dermatology  Volume 112, Issue 1, Pages (January 1999) DOI: /j x Copyright © 1999 The Society for Investigative Dermatology, Inc Terms and Conditions

2 Figure 1 Antigen-pulsed epidermal cells induced proliferation of T cells enriched from lymph node cell suspensions. T cell proliferation was evaluated by [3H]thymidine incorporation. EC, intact epidermal cells; pulsed-EC, pulsed epidermal cells; T, T cell-enriched fraction of lymph node cells from TNCB-sensitized mice; T + TNBS, T cell-enriched fraction of lymph node cells from TNCB-sensitized mice with added antigen; T + EC, T cell-enriched fraction with epidermal cells; T + pulsed-EC, T cell-enriched fraction with pulsed epidermal cells. Data shown are from one representative experiment out of two independent experiments that gave similar results (mean ± SD of triplicate cultures). Journal of Investigative Dermatology  , 19-24DOI: ( /j x) Copyright © 1999 The Society for Investigative Dermatology, Inc Terms and Conditions

3 Figure 2 Dose-dependent suppression of antigen-presenting capacity of epidermal cells by UVA irradiation. UVA (0–20 J per cm2)-irradiated murine epidermal cells were pulsed with TNBS and were cultured with T cells prepared from syngeneic mice sensitized with TNBC. The antigen-presenting function of Langerhans cell is expressed in terms of radioactivity of [3H]thymidine incorporated by proliferating T cells. Data shown are from one representative experiment out of two independent experiments that gave similar results (mean ± SD of triplicate cultures). The asterisk indicates a statistically significant difference (p < 0.05) between the UVA-irradiated cells and sham-irradiated cells. Journal of Investigative Dermatology  , 19-24DOI: ( /j x) Copyright © 1999 The Society for Investigative Dermatology, Inc Terms and Conditions

4 Figure 3 Cell survival of UVA-irradiated epidermal cells. Twenty-four hours after UVA (2.5–10 J per cm2) irradiation, the viability of epidermal cells was assessed by trypan blue exclusion. Survival (%) was determined by use of the following formula: (viability of UVA-irradiated epidermal cells) ÷ (viability of sham-irradiated epidermal cells) × 100. Journal of Investigative Dermatology  , 19-24DOI: ( /j x) Copyright © 1999 The Society for Investigative Dermatology, Inc Terms and Conditions

5 Figure 4 Dose-dependent suppression of upregulation of the ICAM-1, B7–1, and B7–2 expression on Langerhans cell by UVA irradiation. Freshly isolated and cultured epidermal cells were incubated with fluorescein isothiocyanate-conjugated anti-Ia antibody and phycoerythrin-conjugated anti-CD54 antibody for analysis of the ICAM-1 expression (a), and with fluorescein isothiocyanate-conjugated anti-Ia antibody and biotin-conjugated anti-CD80 antibody for analysis of B7–1 (b) or biotin-conjugated anti-CD86 antibody for analysis of B7–2 (c), followed by phycoerythrin-conjugated streptavidin. Expression (%) of these surface molecules was calculated by use of the following formula: (fluorescence intensity of UV-irradiated cultured Langerhans cells – fluorescence intensity of fLangerhans cells) ÷ (fluorescence intensity of sham-irradiated cultured Langerhans cells – fluorescence intensity of fLangerhans cells) × 100. Data were expressed as the mean ± SD of three independent experiments. The asterisk indicates a statistically significant difference (p < 0.05) between the UVA-irradiated cells and sham-irradiated cells. Journal of Investigative Dermatology  , 19-24DOI: ( /j x) Copyright © 1999 The Society for Investigative Dermatology, Inc Terms and Conditions

6 Figure 5 Effect of supernatants from UVA-irradiated epidermal cells on Langerhans cell function. Intact epidermal cells and T cells were cocultured in the supernatants harvested from UVA-irradiated or unirradiated epidermal cells that had been cultured for 1–3 d after irradiation. Data shown are from one representative experiment out of two independent experiments that gave similar results (mean ± SD of triplicate cultures). Journal of Investigative Dermatology  , 19-24DOI: ( /j x) Copyright © 1999 The Society for Investigative Dermatology, Inc Terms and Conditions

7 Figure 6 Protective effect of glutathione against UVA-induced suppression of Langerhans cell function. Epidermal cells suspended in RPMI/10% fetal bovine serum were irradiated with UVA (5 J per cm2) in the presence (0.3–3 mM) or absence of glutathione. Data are from one representative experiment out of two independent experiments (mean ± SD of triplicate cultures). The asterisk indicates a statistically significant difference (p < 0.05) between the glutathione treated cells and untreated cells. Journal of Investigative Dermatology  , 19-24DOI: ( /j x) Copyright © 1999 The Society for Investigative Dermatology, Inc Terms and Conditions

8 Figure 7 Protective effect of glutathione against UVA (2.5–10 J per cm2)-induced suppression of ICAM-1 expression. Epidermal cells were irradiated with UVA (2.5–10 J per cm2) in the presence or absence of 3 mM glutathione. The ICAM-1 expression (%) was calculated as described in the legend to Figure 2 . Data were expressed as the mean ± SD of three independent experiments. The asterisk indicates a statistically significant difference (p < 0.05) between the glutathione treated cells and untreated cells. Journal of Investigative Dermatology  , 19-24DOI: ( /j x) Copyright © 1999 The Society for Investigative Dermatology, Inc Terms and Conditions

9 Figure 8 Dose-dependent protective effect of glutathione against UVA-induced suppression of ICAM-1 expression. Epidermal cells were irradiated with UVA (5 J per cm2) in the presence (0.3–3 mM) or absence of glutathione. [The ICAM-1 expression (%) was calculated as described in the legend to Figure 4. Data are expressed as the mean ± SD of three independent experiments. The asterisk indicates a statistically significant difference (p < 0.05) between the glutathione treated and untreated cells. Journal of Investigative Dermatology  , 19-24DOI: ( /j x) Copyright © 1999 The Society for Investigative Dermatology, Inc Terms and Conditions

10 Figure 9 Protective effect of glutathione against the suppression of endogenous proliferation of lymph node cells induced by in vivo UVA irradiation. The back of mice was exposed to 130 J per cm2 UVA. During irradiation, 15 μl of 3% glutathione solution or vehicle was topically applied every 2 h. TNCB was applied to the irradiated sites on the next day. Radioactivity of [3H]thymidine incorporated by regional lymph node cells was measured 3 d after sensitization. Data are from one representative experiment out of three independent experiments that gave similar results (mean ± SD of triplicate cultures). The asterisk indicates a statistically significant difference (p < 0.05) between the glutathione treated cells and untreated cells. Journal of Investigative Dermatology  , 19-24DOI: ( /j x) Copyright © 1999 The Society for Investigative Dermatology, Inc Terms and Conditions

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