Microvesicles (MVs) are a subtype of extracellular vesicle (EVs) released by cells, which play important roles in cell-cell communication. The role of host cell-derived MVs in host-pathogen interactions remains to be defined in the context of mycobacterial infection. In this study, we characterized MVs that were isolated from mouse RAW 264.7 cells either uninfected or infected with Mycobacterium abscessus (M.ab), one of the most common non-tuberculous mycobacterial (NTM) pathogens in humans, in cell culture. The proteomic profile of purified MVs was determined by mass spectrometry (MS)-based proteomics.