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Quinolone-Photoconjugated Major Histocompatibility Complex Class II-Binding Peptides with Lysine are Antigenic for T Cells Mediating Murine Quinolone.

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Presentation on theme: "Quinolone-Photoconjugated Major Histocompatibility Complex Class II-Binding Peptides with Lysine are Antigenic for T Cells Mediating Murine Quinolone."— Presentation transcript:

1 Quinolone-Photoconjugated Major Histocompatibility Complex Class II-Binding Peptides with Lysine are Antigenic for T Cells Mediating Murine Quinolone Photoallergy  Yoshiki Tokura, Naohiro Seo, Masahiro Takigawa  Journal of Investigative Dermatology  Volume 117, Issue 5, Pages (November 2001) DOI: /j x x Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions

2 Figure 1 Chemical structure of OFLX.
Journal of Investigative Dermatology  , DOI: ( /j x x) Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions

3 Figure 2 The amount of lysine is decreased on amino acid composition analysis of BSA photoreacted with OFLX. A solution of BSA and OFLX was irradiated with UVA and subjected to amino acid analysis. Data are expressed as the percentage reduction in each amino acid relative to untreated BSA. Journal of Investigative Dermatology  , DOI: ( /j x x) Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions

4 Figure 3 The concentration of OFLX is higher in OFLX-H6KH5 than OFLX-H12 photoadducts. UVA-irradiated solutions of OFLX and H12 or H6KH5 were applied to ST-Q-9-Sepharose affinity column. Each fraction of the acid-eluates was measured in the amount of OFLX with a spectrophotometer, and fraction 3 was quantified in the concentration of peptide as estimated by the amount of histidine. Journal of Investigative Dermatology  , DOI: ( /j x x) Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions

5 Figure 4 Proliferation of immune CD4+ T cells in response to Langerhans cells pulsed with OFLX-photoconjugated, I-Ad-binding peptides. CD4+ cells were purified with immunomagnetic beads from LN cells of mice immunized s.c. with OFLX-photomodified epidermal cells. Langerhans cell-enriched epidermal cells from naive mice were precultured with the indicated peptide that were photoconjugated or unconjugated with OFLX. CD4+ cells and Langerhans cell-enriched epidermal cells thus prepared were cocultured for 3 d and the proliferative response of CD4+ cells was measured by 3H-thymidine incorporation. Data are expressed as Δ cpm of OFLX-peptide or peptide responses: (cpm of culture in the presence of Langerhans cells loaded with OFLX-peptide or peptide - cpm of culture in the presence of unloaded Langerhans cells). Error bars represent SD. *p < 0.05, compared with OFLX-unconjugated group. Journal of Investigative Dermatology  , DOI: ( /j x x) Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions

6 Figure 5 Induction of OFLX photoallergy by e.c. application of OFLX-photoconjugated, I-Ad-binding peptides. Mice were sensitized e.c. with OVA9(K+) that were photoconjugated with OFLX (OFLX-peptide), incubated simply without UVA irradiation (OFLX/peptide), or untreated (peptide). As control sensitization, mice received s.c. injections of OFLX-photomodified syngeneic epidermal cells. Six days after immunization, they were administered i.p. with OFLX and irradiated 24 h later with UVA on the earlobes. The ear swelling response was measured 24 h after irradiation. Error bars represent SD. *p < 0.05, compared with no sensitization group. Journal of Investigative Dermatology  , DOI: ( /j x x) Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions

7 Figure 6 In vitro proliferation of CD4+ T cells from mice sensitized e.c. with OFLX-peptide photoadducts in response to OFLX-photoconjugated, I-Ad-binding peptides. OFLX-OVA9(K+) was used as the peptide in this study. CD4+ cells from mice sensitized e.c. with OFLX-peptide photoconjugates or peptide itself were cultured with Langerhans cell-enriched epidermal cells loaded with peptide that were photoconjugated with OFLX (OFLX-peptide), incubated without UVA irradiation (OFLX/peptide), or untreated (peptide). 3H-thymidine incorporation was measured after 72 h cultivation. Error bars represent SD. *p < 0.05, compared with no loading group. **p < 0.05, compared with OFLX/peptide and peptide groups. Journal of Investigative Dermatology  , DOI: ( /j x x) Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions

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