Proteomics and Network Analyses Reveal Inhibition of Akt-mTOR Signaling in CD4⁺ T Cells by Mycobacterium tuberculosis Mannose-Capped Lipoarabinomannan

Authors

Ahmad F. Karim, Case Western Reserve University
Obondo J. Sande, Case Western Reserve UniversityFollow
Sara E. Tomechko, Case Western Reserve University
Xuedong Ding, Case Western Reserve University
Ming Li, Case Western Reserve University
Sean Maxwell, Case Western Reserve UniversityFollow
Clifford V. Harding, Case Western Reserve UniversityFollow
Roxana E. Rojas, Case Western Reserve UniversityFollow
Mark R. Chance, Case Western Reserve UniversityFollow
W. Henry Boom, Case Western Reserve UniversityFollow

Author ORCID Identifier

Clifford V. Harding

Mark R. Chance

Document Type

Article

Publication Date

10-10-2017

DOI

10.1002/pmic.201700233

Abstract

Mycobacterium tuberculosis (Mtb) cell wall glycolipid mannose-capped lipoarabinomannan (ManLAM) inhibits CD4+ T-cell activation by inhibiting proximal T-cell receptor (TCR) signaling when activated by anti-CD3. To understand the impact of ManLAM on CD4+ T-cell function when both the TCR–CD3 complex and major costimulator CD28 are engaged, we performed label-free quantitative MS and network analysis. Mixed-effect model analysis of peptide intensity identified 149 unique peptides representing 131 proteins that were differentially regulated by ManLAM in anti-CD3- and anti-CD28-activated CD4+ T cells. Crosstalker, a novel network analysis tool identified dysregulated translation, TCA cycle, and RNA metabolism network modules. PCNA, Akt, mTOR, and UBC were found to be bridge node proteins connecting these modules of dysregulated proteins. Altered PCNA expression and cell cycle analysis showed arrest at the G2M phase. Western blot confirmed that ManLAM inhibited Akt and mTOR phosphorylation, and decreased expression of deubiquitinating enzymes Usp9x and Otub1. Decreased NF-κB phosphorylation suggested interference with CD28 signaling through inhibition of the Usp9x-Akt-mTOR pathway. Thus, ManLAM induced global changes in the CD4+ T-cell proteome by affecting Akt-mTOR signaling, resulting in broad functional impairment of CD4+ T-cell activation beyond inhibition of proximal TCR–CD3 signaling.

Keywords

Akt, CD4 T-cell +, label-free mass spectrophotometry, M. tuberculosis, ManLAM, mTOR

Publication Title

Proteomics

Volume

17

Issue

22

Rights

© 2017 The Authors, Proteomics Published by WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim

Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 International License.

Recommended Citation

Karim, A. F., Sande, O. J., Tomechko, S. E., Ding, X., Li, M., Maxwell, S., Ewing, R. M., Harding, C. V., Rojas, R. E., Chance, M. R., Boom, W. H., Proteomics 2017, 17, 1700233. https://doi.org/10.1002/pmic.201700233