Revealing immune responses in the Mycobacterium avium subsp. paratuberculosis-infected THP-1 cells using single cell RNA-sequencing

To determine the initial responses of human THP-1 cells against MAP infection using single-cell RNA-seq analysis. Differentiation of THP-1 cells into macrophages was performed by stimulation with 50 ng/ml PMA. After 72 h of incubation, the cells were washed twice with FBS-free RPMI 1640 medium and incubated with 5% FBS-RPMI 1640 medium without antibiotics for 24 h before the experiments. Differentiated THP-1 cells (5 × 106 cells) were inoculated with MAP strain K-10 at a multiplicity of infection (MOI) of 10:1 and incubated for 3 h.