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The treatment of NTM infection in CF patients is challenging due to the natural resistance of NTM species to many antibiotics. <jats:italic>Mycobacterium abscessus<\/jats:italic> is one of the most common NTM species found in the airways of CF patients. In this study, we characterized the extracellular vesicles (EVs) released by drug\u2010sensitive <jats:italic>M. abscessus<\/jats:italic> untreated or treated with clarithromycin (CLR), one of the frontline anti\u2010NTM drugs. Our data show that exposure to CLR increases mycobacterial protein trafficking into EVs as well as the secretion of EVs in culture. Additionally, EVs released by CLR\u2010treated <jats:italic>M. abscessus<\/jats:italic> increase <jats:italic>M. abscessus<\/jats:italic> resistance to CLR when compared to EVs from untreated <jats:italic>M. abscessus<\/jats:italic>. Proteomic analysis further indicates that EVs released by CLR\u2010treated <jats:italic>M. abscessus<\/jats:italic> carry an increased level of 50S ribosomal subunits, the target of CLR. Taken together, our results suggest that EVs play an important role in <jats:italic>M. abscessus<\/jats:italic> resistance to CLR treatment.<\/jats:p>","DOI":"10.1002\/pmic.202300332","type":"journal-article","created":{"date-parts":[[2024,1,18]],"date-time":"2024-01-18T23:51:13Z","timestamp":1705621873000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":12,"title":["<i>Mycobacterium abscessus<\/i> extracellular vesicles increase mycobacterial resistance to clarithromycin in vitro"],"prefix":"10.1002","volume":"24","author":[{"given":"Charlie A.","family":"Vermeire","sequence":"first","affiliation":[{"name":"Department of Biochemistry and Molecular Biology Oklahoma State University  Stillwater Oklahoma USA"},{"name":"Oklahoma Center for Respiratory and Infectious Diseases Oklahoma State University  Stillwater Oklahoma 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