Identification of the mouse Kupffer cell receptors recognizing pneumococcal capsules by affinity screening

Haoran An; Xianbin Tian; Yijia Huang; Jing-Ren Zhang

doi:10.1016/j.xpro.2023.102065

Identification of the mouse Kupffer cell receptors recognizing pneumococcal capsules by affinity screening

STAR Protoc. 2023 Mar 17;4(1):102065. doi: 10.1016/j.xpro.2023.102065. Epub 2023 Jan 24.

Authors

Haoran An¹, Xianbin Tian¹, Yijia Huang¹, Jing-Ren Zhang²

Affiliations

¹ Center for Infectious Disease Research, Department of Basic Medical Science, School of Medicine, Tsinghua University, Beijing, China.
² Center for Infectious Disease Research, Department of Basic Medical Science, School of Medicine, Tsinghua University, Beijing, China; Tsinghua-Peking Center for Life Sciences, Tsinghua University, Beijing, China. Electronic address: zhanglab@mail.tsinghua.edu.cn.

Abstract

Kupffer cells (KCs) are the major sentinels to guard the bloodstream by recognizing diverse microbial ligands of blood-borne pathogens. Here, we establish a protocol for identifying the KC receptors recognizing the capsular polysaccharides (CPSs) of low-virulence Streptococcus pneumoniae in a mouse model. This protocol includes preparation of CPS-coated microspheres and KC membrane proteins, affinity pulldown of CPS-binding proteins, and functional validation of the CPS receptors. This protocol provides a platform to investigate the receptor-ligand interactions between KCs and encapsulated bacteria. For complete details on the use and execution of this protocol, please refer to An et al. (2022).¹.

Keywords: Cell Biology; Flow Cytometry/Mass Cytometry; High Throughput Screening; Immunology; Microbiology; Molecular Biology.

Publication types

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

MeSH terms

Animals
Mice
Streptococcus pneumoniae* / metabolism

Substances

Kupffer cell receptor