Isolation and transcriptional profiling of antigen-presenting cells from mouse lung and MLNs following intranasal immunization

Xiuyu Wang; Yihan Wang; Qiang Wang; Anita S Chong; Shengbo Cao; Youhui Si

doi:10.1016/j.xpro.2024.103400

Isolation and transcriptional profiling of antigen-presenting cells from mouse lung and MLNs following intranasal immunization

STAR Protoc. 2024 Dec 20;5(4):103400. doi: 10.1016/j.xpro.2024.103400. Epub 2024 Oct 19.

Authors

Xiuyu Wang¹, Yihan Wang², Qiang Wang², Anita S Chong³, Shengbo Cao⁴, Youhui Si⁵

Affiliations

¹ National Key Laboratory of Agricultural Microbiology, Hubei Hongshan Laboratory, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China.
² Department of Surgery, The University of Chicago, Chicago, IL 60637, USA.
³ Department of Surgery, The University of Chicago, Chicago, IL 60637, USA. Electronic address: [email protected].
⁴ National Key Laboratory of Agricultural Microbiology, Hubei Hongshan Laboratory, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China. Electronic address: [email protected].
⁵ National Key Laboratory of Agricultural Microbiology, Hubei Hongshan Laboratory, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China. Electronic address: [email protected].

Abstract

Dendritic cells (DCs) play a central role in the initiation of the adaptive immune response. Here, we present a protocol for isolating and transcriptionally profiling antigen-presenting cells (APCs) from the mouse lung and mediastinal lymph nodes (MLNs) following intranasal immunization. We describe steps for preparing single-cell suspensions from the lung and MLN, along with the detection and RNA sequencing (RNA-seq) of antigen-presenting DCs. This protocol offers a broadly applicable approach for identifying variations in DC subpopulations under diverse experimental conditions. For complete details on the use and execution of this protocol, please refer to Youhui et al.¹.

Keywords: Cell isolation; Flow Cytometry; Immunology; Molecular Biology; RNA-Seq.

MeSH terms

Administration, Intranasal*
Animals
Antigen-Presenting Cells* / cytology
Antigen-Presenting Cells* / immunology
Cell Separation / methods
Dendritic Cells / immunology
Gene Expression Profiling / methods
Immunization* / methods
Lung* / cytology
Lung* / immunology
Lymph Nodes* / cytology
Lymph Nodes* / immunology
Mice
Mice, Inbred C57BL