IL-21 Reduces Immediate Hypersensitivity Reactions in Mouse Skin by Suppressing Mast Cell Activation or IgE Production Risa Tamagawa-Mineoka, Tsunao.

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Volume 13, Issue 2, Pages (October 2015)

Presentation transcript:

IL-21 Reduces Immediate Hypersensitivity Reactions in Mouse Skin by Suppressing Mast Cell Activation or IgE Production Risa Tamagawa-Mineoka, Tsunao Kishida, Osam Mazda, Norito Katoh Journal of Investigative Dermatology Volume 131, Issue 7, Pages 1513-1520 (July 2011) DOI: 10.1038/jid.2011.73 Copyright © 2011 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Schemes showing the timing of sensitization, challenge, and recombinant murine (rm)IL-21 administration. (a) Mice were sensitized by intraperitoneal injection of ovalbumin (OVA) and given rmIL-21 or vehicle intraperitoneally twice a week for 2 weeks during the sensitization period. Reactions to OVA were elicited by intracutaneous injection of OVA in the ear on day 14. (b) In separate experiments, mice were sensitized by intraperitoneal injection of OVA and given rmIL-21 or vehicle intraperitoneally once a day on days 14–17. Reactions to OVA were elicited by intracutaneous injection of OVA in the ear at 2hours after the final rmIL-21 administration on day 17. Journal of Investigative Dermatology 2011 131, 1513-1520DOI: (10.1038/jid.2011.73) Copyright © 2011 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Effect on the cutaneous immediate hypersensitivity reaction (IHR) of recombinant murine (rm)IL-21 administration during sensitization. Mice were sensitized by intraperitoneal injection of ovalbumin (OVA) and given rmIL-21 or vehicle intraperitoneally twice a week for 2 weeks during the sensitization period. Reactions to OVA were elicited by intracutaneous injection of OVA in the ear on day 14. Ear swelling responses were examined by measuring the (a) thickness and (b) weight of the ears. (c) Histology of ear skin was examined at 1 and 24hours. Hematoxylin and eosin. Original magnification × 100, scale bar=50μm. (d) Myeloperoxidase (MPO) and eosinophil peroxidase (EPO) activities in skin tissue were examined at 24hours. Data are expressed as means±SD of six mice per group. *P<0.01 versus vehicle administration in OVA-challenged mice. ##P<0.01 versus corresponding sham-challenged mice. Journal of Investigative Dermatology 2011 131, 1513-1520DOI: (10.1038/jid.2011.73) Copyright © 2011 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 Effect on the cutaneous immediate hypersensitivity reaction (IHR) of recombinant murine (rm)IL-21 administration after sensitization. Mice were sensitized by intraperitoneal injection of ovalbumin (OVA) and then given rmIL-21 or vehicle intraperitoneally once a day on days 14–17. Reactions to OVA were elicited by intracutaneous injection of OVA in the ear after the final rmIL-21 administration on day 17. Ear swelling responses were determined by the (a) ear thickness and (b) weight. (c) Histology of ear skin was examined at 1 and 24hours. Hematoxylin and eosin. Original magnification × 100, scale bar=50μm. (d) Myeloperoxidase (MPO) and eosinophil peroxidase (EPO) activities in skin tissue were examined at 24hours. Data are expressed as means±SD of six mice per group. *P<0.01 versus vehicle in OVA-challenged mice. ##P<0.01 versus corresponding sham-challenged mice. Journal of Investigative Dermatology 2011 131, 1513-1520DOI: (10.1038/jid.2011.73) Copyright © 2011 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 Serum levels of allergen-specific IgE and total IgE. (a) Mice were sensitized by intraperitoneal injection of ovalbumin (OVA) and given recombinant murine (rm)IL-21 or vehicle intraperitoneally twice a week for 2 weeks during the sensitization period. Sera were collected on day 14. (b) In separate experiments, mice were sensitized by intraperitoneal injection of OVA and given rmIL-21 or vehicle intraperitoneally once a day on days 14–17. Sera were collected at 2hours after the final rmIL-21 administration on day 17. Serum levels of total IgE and OVA-specific IgE were measured using the corresponding ELISA kits. Data are expressed as means±SD of six mice per group. *P<0.01 versus vehicle in OVA-challenged mice. ##P<0.01 versus corresponding sham-sensitized mice. Journal of Investigative Dermatology 2011 131, 1513-1520DOI: (10.1038/jid.2011.73) Copyright © 2011 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 5 Effect of in vivo recombinant murine (rm)IL-21 administration on mast cell degranulation. (a) Mice were sensitized by injecting ovalbumin (OVA) intraperitoneally and given rmIL-21 or vehicle intraperitoneally once a day on days 14–17. Reactions to OVA were elicited by intracutaneous injection of OVA in the ear of mice at 2hours after the final rmIL-21 administration on day 17. Ears were excised at 1hour after challenge and samples were stained with toluidine blue. Degranulated mast cells are indicated by arrows. Original magnification × 400, scale bar=25μm. (b) Dermal mast cells were evaluated by averaging the number of cells present in 10 high-power fields. Data are expressed as means±SD of six mice per group. *P<0.01 versus vehicle in OVA-challenged mice. #P<0.05 or ##P<0.01 versus corresponding sham-challenged mice. Journal of Investigative Dermatology 2011 131, 1513-1520DOI: (10.1038/jid.2011.73) Copyright © 2011 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 6 Effect of in vitro recombinant murine (rm)IL-21 administration on mediator release from mast cells. (a) Bone marrow-derived mast cells (BMMCs) were labeled by a specific antibody to IL-21 receptor and then IL-21 receptor expression on the surface of BMMCs was analyzed by FACS. Representative data from one of three independent experiments are shown. (b) In separate experiments, BMMCs were sensitized with anti-dinitrophenyl (DNP) IgE, incubated in the presence of rmIL-21 or vehicle, and then challenged with DNP-bovine serum albumin for 0, 0.5, 12, and 24hours. Histamine, leukotriene (LT)C4, tumor necrosis factor-α (TNF-α), and RANTES (regulated on activation, normal T-cell expressed, and presumably secreted) levels were quantified in cell-free supernatants using the corresponding ELISA kits (n=3–5 in each group). *P<0.01 versus vehicle at the same time point after challenge. Journal of Investigative Dermatology 2011 131, 1513-1520DOI: (10.1038/jid.2011.73) Copyright © 2011 The Society for Investigative Dermatology, Inc Terms and Conditions