Mycobacterium abscessus (M.abs) is a rapidly growing mycobacterial species that infects macrophages, and is an important pathogen in patients with
cystic fibrosis. We studied the early stages of M.abs
infection of macrophages, with emphasis on the role of heme-oxygenase-1 (HO-1) in this
infection. THP-1 cells were activated using TPA into macrophage-like cells and infected with M.abs for different time points. M.abs
infection robustly induced HO-1 expression in the THP-1 cells. Production of HO-1 was
p38 MAPK-dependent, as p38 inhibitors suppressed HO-1 induction. Pretreatment with HO-1 inhibitors
tin-protoporphyrin (SnPP) significantly inhibited M.abs growth inside macrophages. Furthermore, inhibiting HO-1 using HO-1
siRNA or the HO-1 upstream signaling molecule; Nrf2 using Nrf2
siRNA resulted in similar inhibition of M.abs. In contrast, inducing HO-1 did not increase M.abs intracellular growth above control. Products of HO-1 metabolism of
heme are
bilirubin,
biliverdin,
carbon monoxide (CO) and
iron. The addition of either
bilirubin or
biliverdin, but not CO, completely restored the SnPP inhibitory effect and partially that with HO-1
siRNA. To understand the mechanisms, we used Syto-62 labeled M.abs to infect macrophages. Interestingly, HO-1 inhibition promoted M.abs-containing phagosome fusion with lysosomes, which should enhance M.abs killing. M.abs
infection enhanced THP-1 ROS production as demonstrated by increased DHE, DCF fluorescence, and EPR signal. HO-1 inhibition further increased ROS production in infected macrophages. Our results indicate that HO-1 induction is important for M.abs growth during the early stages of
infection, and that the HO-1 products
bilirubin and
biliverdin, perhaps through modulation of intracellular ROS levels, may be involved.