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Contact Hypersensitivity in MHC Class II-Deficient Mice Depends on CD8 T Lymphocytes Primed by Immunostimulating Langerhans Cells  Anne Bouloc, Andrea.

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Presentation on theme: "Contact Hypersensitivity in MHC Class II-Deficient Mice Depends on CD8 T Lymphocytes Primed by Immunostimulating Langerhans Cells  Anne Bouloc, Andrea."— Presentation transcript:

1 Contact Hypersensitivity in MHC Class II-Deficient Mice Depends on CD8 T Lymphocytes Primed by Immunostimulating Langerhans Cells  Anne Bouloc, Andrea Cavani, Stephen I. Katz  Journal of Investigative Dermatology  Volume 111, Issue 1, Pages (July 1998) DOI: /j x Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

2 Figure 1 MHC class II-deficient mice have enhanced CHS reactions to TNCB in vivo compared with wild-type littermate controls. (A) Wild-type and MHC class II-deficient mice were sensitized with 100 μl 3% TNCB on the abdomen and challenged 6 d later on the right ear with 20 μl 1% TNCB (four mice per group). The results are expressed as the mean of increase of ear swelling (±SD) 24 h after challenge. The data are representative of two separate experiments. (B) Wild-type and MHC class II-deficient mice were sensitized with 100 μl 3% TNCB on the abdomen and challenged 6 d later on the right ear with 20 μl of different concentrations of TNCB: 1%, 0.5%, 0.1% (four mice per group). The results are expressed as the mean of increase of ear swelling (±SD) 24, 48, and 72 h after challenge. Journal of Investigative Dermatology  , 44-49DOI: ( /j x) Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

3 Figure 2 In vivo depletion with anti-CD8 monoclonal antibody inhibits CHS to TNCB in MHC class II-deficient mice. MHC class II-deficient mice were injected intraperitoneally with 100 μg anti-CD8 monoclonal antibody ( ) or with rat IgG on three consecutive days (four mice per group). One day later the animals were sensitized with 100 μl 3% TNCB on the abdomen. They were then challenged after 6 d on the right ear with 20 μl 1% TNCB. The results are expressed as the mean of increase of ear swelling (±SD) 24 h after challenge. The data are representative of two separate experiments. Journal of Investigative Dermatology  , 44-49DOI: ( /j x) Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

4 Figure 3 Injection of anti-CD8 monoclonal antibodies into wild-type controls results in inhibition of CHS to TNCB and the injection of anti-CD4 antibodies results in enhancement of CHS. Mice were injected intraperitoneally with 100 μg anti-CD8 ( ) or anti-CD4 monoclonal antibodies (GK1.5) or with rat IgG on three consecutive days (four mice per group). One day later the animals were sensitized with 100 μl 3% TNCB on the abdomen. They were then challenged after 6 d on the right ear with 20 μl 1% TNCB. The results are expressed as the mean of increase of ear swelling (±SD) 24 h after challenge. The data are representative of two separate experiments. Journal of Investigative Dermatology  , 44-49DOI: ( /j x) Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

5 Figure 4 Transfer of hapten-specific CD4+ T cells from wild-type mice into MHC class II-deficient mice results in diminution of CHS to TNCB. MHC class II-deficient mice were sensitized with 100 μl 3% TNCB on the abdomen. They were injected intravenously with 2×106 wild-type hapten-specific CD4+ T cells 6 d later and immediately challenged on the right ear with 20 μl 1% TNCB (four mice per group). The results are expressed as the mean of increase of ear swelling (±SD) 24 h after challenge. The data are representative of two separate experiments. Journal of Investigative Dermatology  , 44-49DOI: ( /j x) Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

6 Figure 5 Epidermal cell from MHC class II-deficient mice upregulate B7–1 and B7–2 costimulatory molecules upon short-term culture. Detection of costimulatory molecules B7–1, B7–2, and CD45 on MHC class II-deficient and wild-type epidermal cell suspensions by two color flow cytometry (data are plotted as intensities of FITC fluorescence on the x axis versus phycoerythrin fluorescence on the y axis). Epidermal cells were studied in freshly prepared suspensions (fEC) and after 3 d of culture (cEC). Journal of Investigative Dermatology  , 44-49DOI: ( /j x) Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

7 Figure 6 Epidermal cells from MHC class II-deficient mice exhibit enhanced MHC class I expression upon short-term culture. Detection of MHC class I antigens and CD45 on MHC class II-deficient and wild-type epidermal cell suspensions by two color flow cytometry (data are plotted as intensities of FITC fluorescence on the x axis versus phycoerythrin fluorescence on the y axis). Epidermal cells were studied in freshly prepared suspensions (fEC) and after 3 d of culture (cEC). Journal of Investigative Dermatology  , 44-49DOI: ( /j x) Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

8 Figure 7 Hapten-derivatized epidermal cells from MHC class II-deficient mice can stimulate (A) hapten-specific CD8+ T cells from MHC class II-deficient mice and (B) hapten-specific T cells from wild-type mice. (A) In vivo primed TNP-specific CD8+ T cells were cocultured with fresh epidermal cells, TNP-fresh epidermal cells, cultured epidermal cells, or TNP-cultured epidermal cells. T cell proliferation was determined by the incorporation of [3H]thymidine (1 μCi per well) during the last 16 h of culture. Results are presented as the mean (±SD) of proliferation assays performed in triplicate. The data are representative of two separate experiments. (B) In vivo primed TNP-specific T cells were cocultured with fresh epidermal cells, TNP-fresh epidermal cells, cultured epidermal cells, or TNP-cultured epidermal cells. T cell proliferation was determined by the incorporation of [3H]thymidine (1 μCi per well) during the last 16 h of culture. Results are presented as the mean (±SD) of proliferation assays performed in triplicate. The data are representative of two separate experiments. Journal of Investigative Dermatology  , 44-49DOI: ( /j x) Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

9 Figure 8 Hapten-derivatized epidermal cells from MHC class II-deficient mice induce CHS to TNCB in MHC class II-deficient mice. Mice were sensitized with either 100 μl 3% TNCB on the abdomen or with 2 × 106 TNP-epidermal cells or FITC-epidermal cells injected in the footpad and challenged 6 d later on the right ear with 20 μl 1% TNCB (four mice per group). The results are expressed as the mean of increase of ear swelling (±SD) 24 h after challenge. Journal of Investigative Dermatology  , 44-49DOI: ( /j x) Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

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