V-ATPases are part of the membrane components of pathogen-containing vacuoles,
although their function in intracellular infection remains elusive. In addition
to organelle acidification, V-ATPases are alternatively implicated in membrane
fusion and anti-inflammatory functions controlled by ATP6V0d2, the d subunit
variant of the V-ATPase complex. Therefore, we evaluated the role of ATP6V0d2 in
the biogenesis of pathogen-containing vacuoles using ATP6V0d2 knock-down
macrophages infected with the protozoan parasite Leishmania amazonensis. These
parasites survive within IFNγ/LPS-activated inflammatory macrophages,
multiplying in large/fusogenic parasitophorous vacuoles (PVs) and inducing
ATP6V0d2 upregulation. ATP6V0d2 knock-down decreased macrophage cholesterol
levels and inhibited PV enlargement without interfering with parasite
multiplication. However, parasites required ATP6V0d2 to resist the influx of
oxidized low-density lipoprotein (ox-LDL)-derived cholesterol, which restored PV
enlargement in ATP6V0d2 knock-down macrophages by replenishing macrophage
cholesterol pools. Thus, we reveal parasite-mediated subversion of host V-ATPase
function toward cholesterol retention, which is required for establishing an
inflammation-resistant intracellular parasite niche.
