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UV Exposure Boosts Transcutaneous Immunization and Improves Tumor Immunity: Cytotoxic T-Cell Priming through the Skin  Pamela Stein, Gerd Rechtsteiner,

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Presentation on theme: "UV Exposure Boosts Transcutaneous Immunization and Improves Tumor Immunity: Cytotoxic T-Cell Priming through the Skin  Pamela Stein, Gerd Rechtsteiner,"— Presentation transcript:

1 UV Exposure Boosts Transcutaneous Immunization and Improves Tumor Immunity: Cytotoxic T-Cell Priming through the Skin  Pamela Stein, Gerd Rechtsteiner, Tobias Warger, Tobias Bopp, Thorsten Fuhr, Steve Prüfer, Hans-Christian Probst, Michael Stassen, Peter Langguth, Hansjörg Schild, Markus P. Radsak  Journal of Investigative Dermatology  Volume 131, Issue 1, Pages (January 2011) DOI: /jid Copyright © 2011 The Society for Investigative Dermatology, Inc Terms and Conditions

2 Figure 1 Modulation of T-cell responses in the skin by UV exposure and imiquimod. (a) Mice were sensitized (0.5% dinitrofluorobenzene (DNFB)) after UV-B exposure (90mJcm-2) or imiquimod (2.5mg) as indicated. Antigen challenge was performed 4 days later on the ears (0.2% DNFB). (b, c) Mice were immunized by transcutaneous immunization (TCI) with imiquimod (2.5mg) using the peptide SIINFEKL (100μg) in 50mg cream per application without or after UV-B exposure (90mJcm-2). The frequency of specific CD8+ T cells after 7 days in the spleen was assessed by flow cytometry (b) or by in vivo cytolytic activity (c). A cumulative analysis of four independent experiments, each performed with three mice per group, is shown. *Significant difference to sens./chall., by unpaired Student's t-test. **Significant difference to UV-B by unpaired Student's t-test. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2011 The Society for Investigative Dermatology, Inc Terms and Conditions

3 Figure 2 UV-B exposure before transcutaneous immunization (TCI) with imiquimod induces a cytotoxic T lymphocyte (CTL) memory response. (a) C57BL/6 mice were immunized by TCI with imiquimod in the absence or presence of UV-B light (90mJcm-2) or after intravenous anti-CD40-Ab (100μg) injection (days -3 to 0). The frequency of peptide-specific CD8+ T cells was monitored after 7, 21, and 40 days. (b) After 30 days the in vivo cytolytic activity was assessed after adoptive transfer of peptide loaded and carboxyfluorescein diacetate succinimidyl ester-labeled target cells without prior restimulation. The depicted results are summarized data from at least three independent experiments with three mice per group. *Significant difference by unpaired Student's t-test. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2011 The Society for Investigative Dermatology, Inc Terms and Conditions

4 Figure 3 Low dose of UVB before transcutaneous immunization (TCI) with imiquimod leads to increased immune responses. C57BL/6 mice were immunized and UV-B irradiated as indicated. (a) Dose of UV light was titrated from 0 to 300mJcm-2 and cytolytic activity was assessed. To determine the optimal timing of UV treatment, we irradiated mice 48 or 24hours before and 24 or 24+48hours after immunization. Specific CD8+ T cells (b) and cytolytic activity (c) were quantified by flow cytometric analysis. One representative experiment out of two performed with n=3 mice per group is shown. *A significant difference (P<0.05) by unpaired Student's t-test compared with the no-UV group in a or the -24hour group in b and c. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2011 The Society for Investigative Dermatology, Inc Terms and Conditions

5 Figure 4 Delayed tumor growth and superior tumor protection after UV-B exposure before transcutaneous immunization (TCI). EG.7 cells (4 × 105) were injected subcutaneously into the flank of mice. Immunization treatments were started after 10–12 days after inoculation when tumors became palpable. The mean tumor size with SEM (a) and survival (b) were monitored. Depicted are the cumulative results of four independent experiments with n=37 mice in the TCI group, n=33 in the UV/TCI group, n=33 in the UV group, and n=30 mice in the untreated control group. *A significant difference (P<0.05) between TCI and UV/TCI by Mantel–Cox test for survival analysis or by paired Student's t-test for comparison of tumor growth; NS, not significant. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2011 The Society for Investigative Dermatology, Inc Terms and Conditions

6 Figure 5 CD11c+, but not Langerin+ dendritic cells (DCs), are essential for cytotoxic T lymphocyte (CTL) responses induced by transcutaneous immunization (TCI) with imiquimod. (a, b) Mice were immunized as indicated after FITC painting of the back skin. Lymph nodes were collected after 48hours and analyzed by flow cytometry. CD11c+MHC II+ cells (median, quartiles, range) or Langerin+MHC II+ cells are shown (gated on PI-CD19-CD90- cells). (c) IL-12p35 mRNA induction by qRT-PCR in the draining (filled bars) and nondraining (open bars) lymph nodes after immunizations is shown. (d) The DC phenotype in the draining lymph node 48hours after the indicated treatments is shown (gated on FITC+CD11c+MHC II+ cells). Langerin-DTREGFP mice (e), chimeric mice reconstituted with bone marrow from Langerin-DTREGFP (Langerin-DTREGFP → C57BL/6; f) or CD11c-DTR mice (CD11c-DTR → C57BL/6; g) were used for immunizations. The depicted in vivo cytolytic activity was assessed 7 days later. “Depleted” indicates intraperitoneal diphtheria toxin (DT) injections as described in the Materials and Methods section. The results are representative of at least two independent experiments performed with n=3 mice per group. *A difference (P<0.05) by Mann–Whitney U-test between the draining versus the nondraining lymph nodes. **A difference (P<0.05) by unpaired Student's t-test between groups treated with DT or not. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2011 The Society for Investigative Dermatology, Inc Terms and Conditions

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