Changes in the membrane proteome of exosomes released from human macrophages after infection with Mycobacterium tuberculosis.

ABSTRACT: One of the greatest obstacles to eliminating TB is the lack of predictive and diagnostic biomarkers. Exosomes are a promising alternative source of biomarkers for TB since they can carry mycobacterial antigens. We hypothesize that exosomes from Mtb-infected macrophages exhibit a characteristic selection of human protein that can be evaluated as TB biomarkers. We looked for the localization—exosomal membrane or lumen—of the differentially abundant proteins. Exosomes were obtained from THP-1-derived macrophages (Mtb infected and uninfected controls). The exosome population was validated by nanoparticle tracking analysis and western blot. The protein composition of exosomes was evaluated by tandem mass spectrometry. Differences in protein composition and abundances between exosomes from infected and control cells were evaluated by t-test the proteomics findings were confirmed by western blot. Using a biotinylation strategy we verified the protein localization. Forty-seven proteins were significantly more abundant in exosomes from Mtb-infected cells, 66% were predicted to be membrane associated. The biotinylation pattern confirmed the membrane-association of some of these proteins.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture, Macrophage

SUBMITTER: Gustavo Diaz  

LAB HEAD: Karen Dobos

PROVIDER: PXD004062 | Pride | 2016-12-20

REPOSITORIES: Pride