Multiparameter flow cytometry assay to analyze the pulmonary T cell profiles in the ovine model of respiratory syncytial virus infection

Thomas Démoulins; Melanie Brügger; Beatrice Zumkehr; Blandina I Oliveira Esteves; Nicolas Ruggli; Marco P Alves

doi:10.1016/j.xpro.2022.101688

Multiparameter flow cytometry assay to analyze the pulmonary T cell profiles in the ovine model of respiratory syncytial virus infection

STAR Protoc. 2022 Dec 16;3(4):101688. doi: 10.1016/j.xpro.2022.101688. Epub 2022 Sep 16.

Authors

Thomas Démoulins¹, Melanie Brügger², Beatrice Zumkehr², Blandina I Oliveira Esteves², Nicolas Ruggli², Marco P Alves³

Affiliations

¹ Institute of Virology and Immunology, Bern, Switzerland; Institute of Veterinary Bacteriology, University of Bern, Bern, Switzerland; Department of Infectious Diseases and Pathobiology, Vetsuisse Faculty, University of Bern, Bern, Switzerland. Electronic address: thomas.demoulins@vetsuisse.unibe.ch.
² Institute of Virology and Immunology, Bern, Switzerland; Department of Infectious Diseases and Pathobiology, Vetsuisse Faculty, University of Bern, Bern, Switzerland.
³ Institute of Virology and Immunology, Bern, Switzerland; Department of Infectious Diseases and Pathobiology, Vetsuisse Faculty, University of Bern, Bern, Switzerland. Electronic address: marco.alves@vetsuisse.unibe.ch.

Abstract

Here, we present a protocol to analyze the T cell profiles of the neonatal ovine lung during respiratory syncytial virus (RSV) infection. The protocol delivers standardized multiparameter flow cytometry (FCM) analysis of CD4⁺, CD8⁺, regulatory, and γδ T cells isolated from lung, lymph nodes, and bronchoalveolar lavages (BALs). We detail the preparation of RSV and transtracheal inoculation of newborn lambs. We then describe tissue isolation and preparation of cell suspensions, followed by FCM acquisition to identify different T cell subsets. For complete details on the use and execution of this protocol, please refer to Démoulins et al. (2021).

Keywords: Flow cytometry/Mass cytometry; Health sciences; Immunology; Microbiology; Model organisms.

Publication types

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

MeSH terms

Animals
Flow Cytometry
Lung / pathology
Respiratory Syncytial Virus Infections* / pathology
Respiratory Syncytial Viruses
Sheep
T-Lymphocyte Subsets