THP1 proteomics in response to mycobacterium tuberculosis infection

Abstract

Temporal data on how the mycobacterium infection establishes itself inside the host cell is not available. We differentiated human THP1 cell line with PMA and infected them with different laboratory (H37Ra and H37Rv) and clinical strains (BND433 and JAL2287) of mycobacterium tuberculosis (Mtb). Uninfected differentiated THP1 cells were used as infection control. Host proteome was investigated at four different time points to understand the dynamics of host response to mycobacterial infection with time. The investigated time points included 6 hrs, 18 hrs, 30 hrs and 42 hrs of infection with all the Mtb strains. SWATH-MS method was used to quantitate the host proteome in response to Mtb infection and the data thus obtained are available via PRIDE repository with the dataset identifier PXD022352 ( https://www.ebi.ac.uk/pride/archive/projects/PXD022352 ).

Temporal Profiling of Host Proteome against Different M. tuberculosis Strains Reveals Delayed Epigenetic Orchestration

“…Different strains of MTB were cultured in Middlebrook 7H9 broth (Becton Dickinson Difco, Sparks, MD, USA) supplemented with 10% ADC (Becton Dickinson, Sparks, MD USA), 0.4% glycerol, and 0.05% Tween 80. Bacteria were harvested in the mid-log phase, resuspended in RPMI medium, dispersed by aspiration to break clumps, and quantified by the spectrophotometric method, as reported before [15]. For THP1 cell lines, RPMI 1640 (Gibco, Grand Island, NY, USA) supplemented with 10% FCS (Hyclone, Marlborough, MA, USA) and 1× penicillin (stock concentration = 100 U/mL) and streptomycin (stock concentration = 100 µg/mL) was used.…”

“…Cells were grown and maintained at 37 • C with appropriate humidity and 5% CO 2 . A detailed methodology can be found in our earlier report [15].…”