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Published on: October 20, 2020
Comparative Proteomics Analysis of Human Macrophages Infected with Virulent Mycobacterium bovis
Pei Li1, Rui Wang1, Wenqi Dong1
1State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural UniversityWuhan, China; Key Laboratory of Development of Veterinary Diagnostic Products, Ministry of Agriculture, Huazhong Agricultural UniversityWuhan, China.
Abstract:
Mycobacterium bovis (M. bovis), the most common pathogens of tuberculosis (TB), is virulent to human and cattle, and transmission between cattle and humans warrants reconsideration concerning food safety and public health. Recently, efforts have begun to analyze cellular proteomic responses induced by Mycobacterium tuberculosis (M. tb). However, the underlying mechanisms by which virulent M. bovis affects human hosts are not fully understood. For the present study, we utilized a global and comparative labeling strategy of isobaric tag for relative and absolute quantitation (iTRAQ) to assess proteomic changes in the human monocyte cell line (THP-1) using a vaccine strain and two virulent strains H37Rv and M. bovis. We measured 2,032 proteins, of which 61 were significantly differentially regulated. Ingenuity Pathway Analysis was employed to investigate the canonical pathways and functional networks involved in the infection. Several pathways, most notably the phagosome maturation pathway and TNF signaling pathway, were differentially affected by virulent strain treatment, including the key proteins CCL20 and ICAM1. Our qRT-PCR results were in accordance with those obtained from iTRAQ. The key enzyme MTHFD2, which is mainly involved in metabolism pathways, as well as LAMTOR2 might be effective upon M. bovis infection. String analysis also suggested that the vacuolar protein VPS26A interacted with TBC1D9B uniquely induced by M. bovis. In this study, we have first demonstrated the application of iTRAQ to compare human protein alterations induced by virulent M. bovis infections, thus providing a conceptual understanding of mycobacteria pathogenesis within the host as well as insight into preventing and controlling TB in human and animal hosts' transmission.
Insights
Mycobacterium bovis (M. bovis) causes tuberculosis (TB) in humans and cattle. This study used iTRAQ to analyze proteomic changes in human cells infected with M. bovis, revealing key pathways and proteins involved in pathogenesis.
Area of Science:
- Proteomics
- Immunology
- Microbiology
Background:
- Tuberculosis (TB) is a significant public health concern, with Mycobacterium bovis (M. bovis) posing risks to both human and cattle populations.
- While Mycobacterium tuberculosis (M. tb) proteomic responses are studied, the mechanisms of M. bovis virulence in human hosts remain unclear.
Purpose of the Study:
- To investigate the proteomic alterations in human monocytes upon infection with virulent M. bovis strains.
- To identify key pathways and proteins involved in the host response to M. bovis.
Main Methods:
- Utilized isobaric tag for relative and absolute quantitation (iTRAQ) for global proteomic analysis.
- Employed Ingenuity Pathway Analysis (IPA) and String analysis for pathway and network analysis.
- Validated findings using quantitative reverse transcription PCR (qRT-PCR).
Main Results:
- Identified 61 significantly differentially regulated proteins out of 2,032 quantified.
- Observed differential regulation in phagosome maturation and TNF signaling pathways.
- Highlighted key proteins such as CCL20, ICAM1, MTHFD2, LAMTOR2, VPS26A, and TBC1D9B.
Conclusions:
- This study is the first to apply iTRAQ to compare human protein alterations induced by virulent M. bovis.
- Provides insights into M. bovis pathogenesis and potential targets for controlling TB transmission between animals and humans.
