STAR Protocols
Volume 3, Issue 4, 16 December 2022, 101688
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Protocol
Multiparameter flow cytometry assay to analyze the pulmonary T cell profiles in the ovine model of respiratory syncytial virus infection

https://doi.org/10.1016/j.xpro.2022.101688Get rights and content
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Highlights

  • Procedure to generate a translational neonatal model of RSV disease

  • Approach to assess T cell responses from lung, lymph nodes, and BAL of newborn lambs

  • Steps to isolate tissue and prepare cell suspensions

  • Standardized multiparameter FCM assay to identify different T cell subsets

Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Summary

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).

Subject areas

Flow cytometry/Mass cytometry
Health sciences
Immunology
Microbiology
Model organisms

Data and code availability

No data or code was generated or analyzed in this protocol.

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