Tirap controls Mycobacterium tuberculosis phagosomal acidification
Fig 5
Tirap implication in Mtb effectors secretion.
(A) BMDMs were seeded into 96-well plates and loaded with homologous (pos) or control (-) peptides or infected with Mtb H37Rv-DsRed. 24 hours post incubation, cells were washed and co-cultured with transduced anti-Ag85 or anti-Esat6 reporter T-cell hybridomas. These were then analyzed by automated confocal microscopy. Shown are representative images of DAPI labelled T-cells (blue) and Antigen specific T-cells (green). Histograms showing mean ± SEM of percentage of ESAT-6 or Ag85 specific T-cells obtained from 5 analysed wells. (B) BMDMs from WT, Tirap+/- and Tirap-/- mice were infected with an attenuated strain of Mtb (H37Ra-GFP) (MOI of 2) and analyzed by automated confocal microscopy. The histogram shows the replication fold increase from 3 hpi to 4 dpi. Shown are mean ± SEM obtained from at least 12 analyzed wells per condition. (C) WT (+/+), knocked-out (-/-) for Myd88, TLR2, TLR4 and TLR9 BMDMs were infected with H37Rv-GFP (MOI of 2) and analyzed by automated confocal microscopy. The histogram shows the replication fold increase from 3 hpi to 4 dpi. Shown are mean ± SEM obtained from at least 4 analyzed wells per condition of one representative out of two independent experiments. *** P value < 0.001 as determined by one-way ANOVA test.