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Updated: Jun 9, 2026
The Utilization of Oropharyngeal Intratracheal PAMP Administration and Bronchoalveolar Lavage to Evaluate the Host Immune Response in Mice
Published on: April 3, 2014
Protocol for visualization and quantitative analysis of murine lung immunological architecture using iBALT-UNMAP
Nicholas J First1, Katelyn M Parrish2, Jose Pedreira-Lopez2
1Department Microbiology and Immunology, Louisiana State University Health Sciences Center at Shreveport, Shreveport, LA 71103, USA; Department of Microbiology, Immunology, and Molecular Genetics, UT Health San Antonio, San Antonio, TX 78229, USA.
Abstract:
The spatial distribution of adaptive immune cells is a critical determinant of the quality and magnitude of mucosal immunity. Here, we present a protocol for tyramide signal amplification (TSA) immunofluorescence that preserves the lung architecture and quantifies inducible bronchus-associated lymphoid tissue (iBALT) following murine Bordetella bronchiseptica infection. Starting from paraffin-embedded lungs, we describe steps for staining, imaging, and morphometric analysis. This protocol can be optimized for other pathogens and experimental settings to allow mapping of spatial immune responses within the respiratory mucosa. For complete details on the use and execution of this protocol, please refer to First et al.1.
