Abstract Mycobacterium abscessus (Mab) colonies adopt smooth (S) or rough (R) morphotypes, which are linked to the presence or absence of glycopeptidolipids (GPL), respectively. Though clinically relevant, the association between GPL levels, morphotype and pathogenesis are poorly understood. To investigate the degree of correlation between Mab morphology, GPL levels, and infectivity, we generated isolates from Mab-positive sputum samples from cystic fibrosis patients. Isolated strains were categorised based on their morphology, GPL profile, and replication rate in macrophages. Our findings revealed that around 50% of isolates displayed mixed morphologies and GPL analysis confirmed a consistent relationship between GPL content and morphotype was only found in smooth isolates. Across morphotype groups, no differences were observed in vitro , yet using a high-content THP-1 cell ex vivo infection model, clinical R strains were observed to replicate at higher levels. Moreover, the proportion of infected macrophages was notably higher among clinical R strains compared to their S counterparts at 72 hours post-infection. Clinical variants also infected at significantly higher rates compared to laboratory strains, highlighting the limited translatability of lab strain infection data to clinical contexts. Our study confirmed the general correlation between morphotype and GPL levels in smooth strains yet unveiled more variability within morphotype groups than previously recognised, particularly during intracellular infection. As the rough morphotype is of highest clinical concern, these findings contribute to the expanding knowledge base surrounding Mab infections, offering insights that can steer diagnostic methodologies, and treatment approaches.

The diversity of clinicalMycobacterium abscessusisolates in morphology, glycopeptidolipids and infection rates in a macrophage model