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

Michael R D'Agostino; Sam Afkhami; Alisha Kang; Art Marzok; Matthew S Miller; Zhou Xing

doi:10.1016/j.xpro.2022.101652

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

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

Authors

Michael R D'Agostino¹, Sam Afkhami², Alisha Kang³, Art Marzok⁴, Matthew S Miller⁴, Zhou Xing³

Affiliations

¹ McMaster Immunology Research Centre, M.G. DeGroote Institute for Infectious Disease Research, Department of Biochemistry & Biomedical Sciences, McMaster University, Hamilton, ON L8S 4K1, Canada. Electronic address: dagostim@mcmaster.ca.
² McMaster Immunology Research Centre, M.G. DeGroote Institute for Infectious Disease Research, Department of Medicine, McMaster University, Hamilton, ON L8S 4K1, Canada. Electronic address: afkhams@mcmaster.ca.
³ McMaster Immunology Research Centre, M.G. DeGroote Institute for Infectious Disease Research, Department of Medicine, McMaster University, Hamilton, ON L8S 4K1, Canada.
⁴ McMaster Immunology Research Centre, M.G. DeGroote Institute for Infectious Disease Research, Department of Biochemistry & Biomedical Sciences, McMaster University, Hamilton, ON L8S 4K1, Canada.

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).

Keywords: Cell isolation; Flow Cytometry/Mass Cytometry; Immunology.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
COVID-19* / prevention & control
Immunity, Innate
Lung
Memory T Cells*
Mice
SARS-CoV-2
Vaccination / methods

Grants and funding

CIHR/Canada