Work

Protocol for mapping T cell activation using single-cell RNA-seq

Year: 2024

Type: article

Abstract: Stimulation of CD4 T cells with anti-CD3/CD28 is a commonly used model to study T cell activation. Here, we present a protocol for investigating T cell activation based on anti-CD3/CD28 bead stimulation and single-cell RNA sequencing (scRNA-seq). We describe the workflow from the isolation of human peripheral blood mononuclear cells (PMBCs) and CD4 T cell enrichment to anti-CD3/CD28 bead stimulation, scRNA-seq, and data analysis. For complete details on the use and execution of this protocol, please refer to Li et al. 1 • Steps for CD4 T cell isolation from human whole blood using a density gradient medium • Procedures for anti-CD3/CD28 microbead-based CD4 T cell activation • Instructions for single-cell RNA library construction • Guidance for data integration and downstream analysis of scRNA-seq data Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Stimulation of CD4 T cells with anti-CD3/CD28 is a commonly used model to study T cell activation. Here, we present a protocol for investigating T cell activation based on anti-CD3/CD28 bead stimulation and single-cell RNA sequencing (scRNA-seq). We describe the workflow from the isolation of human peripheral blood mononuclear cells (PMBCs) and CD4 T cell enrichment to anti-CD3/CD28 bead stimulation, scRNA-seq, and data analysis.

Source: STAR Protocols

DOI: 10.1016/j.xpro.2024.103409