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Animal Models of Allergic Disease pp 115–132Cite as

Generation and Characterization of Inducible Lung and Skin-Specific IL-22 Transgenic Mice

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 2223))

Abstract

IL-22 is an IL-10 family cytokine that is increased in asthma and atopic dermatitis (AD). However, the specific role of IL-22 in the pathogenesis of allergic lung inflammation and AD in vivo has yet to be elucidated. We aimed to develop mouse models of allergic diseases in the lung and skin with inducible and tissue-specific expression of IL-22, using a tetracycline (Tet)-controlled system. In this chapter, we describe a series of protocols we have developed to generate a construct that contains the TRE-Tight promoter and mouse IL-22 cDNA based on this system. Furthermore, we describe how to generate TRE-Tight-IL-22 mice through pronuclear microinjection. In our approach, two Tet-on (CC10-rtTA or SPC-rtTA) and a Tet-off (K5-tTA) transgenic mouse lines are selected to crossbreed with TRE-Tight-IL-22 mice to generate inducible tissue-specific transgenic lines. The transgenic strains, CC10-rtTA/TRE-Tight-IL-22 (CC10-rtTA-IL-22) or SPC-rtTA/TRE-Tight-IL-22 (SPC-rtTA-IL-22) mice, do not produce detectable levels of IL-22 in their bronchoalveolar lavage (BAL) samples in the absence of doxycycline (Dox). However, oral Dox treatment of these mice induces IL-22 expression in the BAL, and the airway and lung epithelial cells. For K5-tTA/TRE-Tight-IL-22 (K5-tTA–IL-22) mice, to avoid potential IL-22 toxicity to mouse embryos, Dox is given starting at the time of breeding to suppress tTA and to keep the IL-22 transgene off until the K5-tTA–IL-22 mice are 6 weeks old. Experiments are then initiated by withdrawing Dox from the drinking water. In all cases, IL-22 protein can be detected by immunohistochemistry in the skin of Tg(+) animals, but not in the skin of Tg(−) animals. Utilizing transgenic technology based on the Tetracycline-controlled system, we have established inducible transgenic mouse models in which cytokine IL-22 can be expressed specifically in the lung or skin. These models are valuable for studies in vivo in a broad range of diseases involving IL-22 and will provide a new platform for research and for seeking novel therapeutics in the fields of inflammation, asthma, and allergic dermatitis.

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Acknowledgments

This work was supported by NIH grant AI55064, AAAAI Women’s Award, and AAAAI Faculty K to R Award to T.Z., NIH grant HL079349 to Z.Z., and NSFC grant 81301428 to L.Z.

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Authors and Affiliations

  1. Animal Bio-Safety Level III Laboratory, State Key Laboratory of Virology, Wuhan University School of Medicine, Wuhan, China

    Li Zhou

  2. Department of Molecular Microbiology and Immunology and Department of Pediatrics, Brown University Warren Alpert Medical School, Providence, RI, USA

    Tao Zheng & Zhou Zhu

Authors
  1. Li Zhou
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  2. Tao Zheng
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  3. Zhou Zhu
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Correspondence to Zhou Zhu .

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Editors and Affiliations

  1. Department of Biomedical Sciences, University of North Dakota School of Medicine & Health Sciences, Grand Forks, ND, USA

    Kumi Nagamoto-Combs

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Zhou, L., Zheng, T., Zhu, Z. (2021). Generation and Characterization of Inducible Lung and Skin-Specific IL-22 Transgenic Mice. In: Nagamoto-Combs, K. (eds) Animal Models of Allergic Disease. Methods in Molecular Biology, vol 2223. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1001-5_9

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  • DOI: https://doi.org/10.1007/978-1-0716-1001-5_9

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1000-8

  • Online ISBN: 978-1-0716-1001-5

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