Glycopeptidolipid glycosylation controls surface properties and pathogenicity in Mycobacterium abscessus

Human IRF1 governs macrophagic IFN-γ immunity to mycobacteria

“…Cells were acquired on a Beckman Coulter Gallios flow cytometer and analyzed with FlowJo Software. The infection of MDMs (differentiation with M-CSF and IL-4 (#216-MC-010 and #204-IL-010, respectively, R&D Systems)) with M. abscessus -tdTomato 203 and intracellular CFU determination were performed as previously described 204 . Infected MDMs were acquired on a Beckton Dickinson Fortessa flow cytometer and analyzed with FlowJo Software.…”

Mycobacterium abscessus alkyl hydroperoxide reductase C promotes cell invasion by binding to tetraspanin CD81

“…Despite numerous attempts, we were unable to generate an ahpC deletion mutant using the unmarked deletion strategy, previously validated for the deletion of non-essential genes in Mab . 8 , 38 , 39 Consequently, to investigate the contribution of AhpC in bacterial uptake by macrophages, we took advantage of the tetracycline repressor system that selectively and ectopically controls gene expression in mycobacteria. 40 , 41 To generate a conditional ahpC mutant, we introduced the ahpC minigene with a C-terminally positioned HA epitope tag into Mab ( Figure 7 A).…”

“… 7 Endothelial epithelial cells, fibroblasts, antigen-presenting cells such as macrophages, dendritic cells, and neutrophils have been identified as potential Mab host cells. 8 , 9 , 10 , 11 , 12 , 13 , 14 Mab is found in macrophages where it manages to multiply, in contrast to other fast-growing NTM or non-pathogenic species, which are naturally cleared by macrophages. 15 , 16 , 17 The infection process begins when bacterial products, known as pathogen-associated molecular patterns (PAMP), engage with pattern recognition receptor (PRR), such as Toll-like receptors (TLR) or C-type lectins present in the plasma membrane.…”

The ESX-4 substrates, EsxU and EsxT, modulate Mycobacterium abscessus fitness

“…Disruption of the esxUT gene unmarked in M. abscessus was performed as described previously [46]. Briefly, the pUX1-katG suicide vector was used to generate unmarked double deletion mutants in M. abscessus CIP104536 T (S).…”

“…Complementation was performed after amplifying and cloning esxUT in fusion with an HA-tagging sequence under the control of the hsp60 promoter into the integrative plasmid pMV306 as described [46]. All primers used for the construction of the two mutants are listed in S1 Table.…”