To elucidate how different immune cells contribute to control or progression of M. tuberculosis (Mtb) infection, we developed a technique to perform multi-modal single-cell RNA sequencing (scRNA-seq) from in vivo Mtb-infected lung macrophages. This protocol simultaneously acquires the transcriptome, surface marker expression, and bacterial phenotype of each infected cell. We describe steps for sorting Mtb-infected cells and staining with CITE-seq antibodies, as well as for methanol fixation and generation of scRNA-seq libraries. This protocol can be used on tissues derived from murine, nonhuman primate, and human infections. For complete details on the use and execution of this protocol, please refer to Pisu et al. (2021).1.
Keywords:Bioinformatics; Cell Biology; Flow Cytometry/Mass Cytometry; Genomics; Immunology; Microbiology; Sequencing; Single Cell.