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经鼻腔免疫接种后小鼠肺组织驻留记忆 T 细胞和气道训练性先天免疫的分离和鉴定方案。

Protocol for isolation and characterization of lung tissue resident memory T cells and airway trained innate immunity after intranasal vaccination in mice.

机构信息

McMaster Immunology Research Centre, M.G. DeGroote Institute for Infectious Disease Research, Department of Biochemistry & Biomedical Sciences, McMaster University, Hamilton, ON L8S 4K1, Canada.

McMaster Immunology Research Centre, M.G. DeGroote Institute for Infectious Disease Research, Department of Medicine, McMaster University, Hamilton, ON L8S 4K1, Canada.

出版信息

STAR Protoc. 2022 Aug 29;3(3):101652. doi: 10.1016/j.xpro.2022.101652. eCollection 2022 Sep 16.

Abstract

Vaccination route dictates the quality and localization of immune responses within tissues. Intranasal vaccination seeds tissue-resident adaptive immunity, alongside trained innate responses within the lung/airways, critical for superior protection against SARS-CoV-2. This protocol encompasses intranasal vaccination in mice, step-by-step bronchoalveolar lavage for both cellular and acellular airway components, lung mononuclear cell isolation, and detailed flow cytometric characterization of lung tissue-resident memory T cell responses, and airway macrophage-trained innate immunity. For complete details on the use and execution of this protocol, please refer to Afkhami et al. (2022).

摘要

接种途径决定了组织内免疫应答的质量和定位。鼻内接种可在肺部/气道内引发组织驻留适应性免疫和训练有素的固有免疫反应,这对于抵抗 SARS-CoV-2 至关重要。本方案包括在小鼠中进行鼻内接种,逐步进行支气管肺泡灌洗以获取细胞和无细胞气道成分、肺单核细胞分离以及详细的流式细胞术分析肺组织驻留记忆 T 细胞应答和气道巨噬细胞训练固有免疫。有关该方案使用和实施的完整详细信息,请参见 Afkhami 等人(2022 年)。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2e14/9441341/b41d1a238d62/fx1.jpg

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