Department of Biological Regulation, Weizmann Institute of Science, Rehovot, Israel.
Methods Mol Biol. 2022;2427:133-147. doi: 10.1007/978-1-0716-1971-1_12.
Complex interactions between diverse host immune cells can determine the outcome of pathogen infections. Advances in single-cell RNA sequencing (scRNA-seq) allow detection of the transcriptional patterns of different immune cells at steady state and after infection. To reveal the complex interactions of the human immune system in response to diverse intracellular pathogens, we developed a protocol for scRNA-seq of ex vivo infected human peripheral blood mononuclear cells (PBMCs). We demonstrate here infection with Salmonella enterica serovar Typhimurium, but this protocol can be used for any other pathogen of interest, and expand our knowledge of human host-pathogen biology.
不同宿主免疫细胞之间的复杂相互作用可以决定病原体感染的结果。单细胞 RNA 测序 (scRNA-seq) 的进展可以在稳态和感染后检测不同免疫细胞的转录模式。为了揭示人类免疫系统对不同细胞内病原体的复杂反应,我们开发了一种用于体外感染人外周血单核细胞 (PBMC) 的 scRNA-seq 方案。我们在这里证明了感染鼠伤寒沙门氏菌血清型 Typhimurium,但该方案可用于任何其他感兴趣的病原体,并扩展了我们对人类宿主-病原体生物学的认识。