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Proteomics and network analyses reveal inhibition of Akt-mTOR signaling in CD4+ T cells by mycobacterium tuberculosis Mannose-capped lipoarabinomannan

Proteomics and network analyses reveal inhibition of Akt-mTOR signaling in CD4+ T cells by mycobacterium tuberculosis Mannose-capped lipoarabinomannan
Proteomics and network analyses reveal inhibition of Akt-mTOR signaling in CD4+ T cells by mycobacterium tuberculosis Mannose-capped lipoarabinomannan
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.
Animals, CD4-Positive T-Lymphocytes, Cell Cycle, Female, Gene Regulatory Networks, Lipopolysaccharides, Mannose, Mass Spectrometry, Mice, Mice, Inbred C57BL, Mycobacterium tuberculosis, Oncogene Protein v-akt, Proteomics, Signal Transduction, TOR Serine-Threonine Kinases, Journal Article
1615-9853
1-13
Karim, Ahmad
2a648c9a-d709-4592-b02e-1141e2d6c0cc
Sande, Obondo J.
001474ef-dbff-4720-8476-396dd784a979
Tomechko, Sara E.
baee8bc4-bc2b-42e4-ac4f-bec89fd00e26
Ding, Xuedong
ecc9d34f-200a-427d-91e7-fbff454f00fb
Li, Ming
a0de216a-0965-43a0-9f93-47ef531a850c
Maxwell, Sean
fd5cddb6-5fd3-4f1b-bb7f-bfa60113c3c2
Ewing, Robert
022c5b04-da20-4e55-8088-44d0dc9935ae
Harding, Clifford V.
8f90adb5-e85b-415b-aaf5-f4e3f90ed5d6
Rojas, Roxana E.
08cbdf50-1f4e-4132-b322-5ee0b82b276c
Chance, Mark R.
855fab90-5057-490a-887f-d7c1db99faf5
Boom, W. Henry
cc7e691a-7784-41d6-b55a-29cc0c54f4e0
Karim, Ahmad
2a648c9a-d709-4592-b02e-1141e2d6c0cc
Sande, Obondo J.
001474ef-dbff-4720-8476-396dd784a979
Tomechko, Sara E.
baee8bc4-bc2b-42e4-ac4f-bec89fd00e26
Ding, Xuedong
ecc9d34f-200a-427d-91e7-fbff454f00fb
Li, Ming
a0de216a-0965-43a0-9f93-47ef531a850c
Maxwell, Sean
fd5cddb6-5fd3-4f1b-bb7f-bfa60113c3c2
Ewing, Robert
022c5b04-da20-4e55-8088-44d0dc9935ae
Harding, Clifford V.
8f90adb5-e85b-415b-aaf5-f4e3f90ed5d6
Rojas, Roxana E.
08cbdf50-1f4e-4132-b322-5ee0b82b276c
Chance, Mark R.
855fab90-5057-490a-887f-d7c1db99faf5
Boom, W. Henry
cc7e691a-7784-41d6-b55a-29cc0c54f4e0

Karim, Ahmad, Sande, Obondo J., Tomechko, Sara E., Ding, Xuedong, Li, Ming, Maxwell, Sean, Ewing, Robert, Harding, Clifford V., Rojas, Roxana E., Chance, Mark R. and Boom, W. Henry (2017) Proteomics and network analyses reveal inhibition of Akt-mTOR signaling in CD4+ T cells by mycobacterium tuberculosis Mannose-capped lipoarabinomannan. Proteomics, 17 (22), 1-13. (doi:10.1002/pmic.201700233).

Record type: Article

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.

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More information

e-pub ahead of print date: 21 November 2016
Published date: November 2017
Keywords: Animals, CD4-Positive T-Lymphocytes, Cell Cycle, Female, Gene Regulatory Networks, Lipopolysaccharides, Mannose, Mass Spectrometry, Mice, Mice, Inbred C57BL, Mycobacterium tuberculosis, Oncogene Protein v-akt, Proteomics, Signal Transduction, TOR Serine-Threonine Kinases, Journal Article

Identifiers

Local EPrints ID: 420343
URI: http://eprints.soton.ac.uk/id/eprint/420343
ISSN: 1615-9853
PURE UUID: a94a5a58-07c3-45b2-967b-b4d99511b6de
ORCID for Robert Ewing: ORCID iD orcid.org/0000-0001-6510-4001

Catalogue record

Date deposited: 04 May 2018 16:30
Last modified: 10 Nov 2021 03:31

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Contributors

Author: Ahmad Karim
Author: Obondo J. Sande
Author: Sara E. Tomechko
Author: Xuedong Ding
Author: Ming Li
Author: Sean Maxwell
Author: Robert Ewing ORCID iD
Author: Clifford V. Harding
Author: Roxana E. Rojas
Author: Mark R. Chance
Author: W. Henry Boom

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