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Updated: May 26, 2026
Microscopy-based Assays for High-throughput Screening of Host Factors Involved in Brucella Infection of Hela Cells
Published on: August 5, 2016
Deep-sequencing analysis of the mouse transcriptome response to infection with Brucella melitensis strains of
Fangkun Wang1, Sen Hu, Wenxing Liu
1State Key Laboratory of Veterinary Biotechnology and Zoonosis Laboratory of the Ministry of Agriculture, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, Heilongjiang, People's Republic of China.
Abstract:
Brucella melitensis is an important zoonotic pathogen that causes brucellosis, a disease that affects sheep, cattle and occasionally humans. B. melitensis strain M5-90, a live attenuated vaccine cultured from B. melitensis strain M28, has been used as an effective tool in the control of brucellosis in goats and sheep in China. However, the molecular changes leading to attenuated virulence and pathogenicity in B. melitensis remain poorly understood. In this study we employed the Illumina Genome Analyzer platform to perform genome-wide digital gene expression (DGE) analysis of mouse peritoneal macrophage responses to B. melitensis infection. Many parallel changes in gene expression profiles were observed in M28- and M5-90-infected macrophages, suggesting that they employ similar survival strategies, notably the induction of anti-inflammatory and antiapoptotic factors. Moreover, 1019 differentially expressed macrophage transcripts were identified 4 h after infection with the different B. melitensis strains, and these differential transcripts notably identified genes involved in the lysosome and mitogen-activated protein kinase (MAPK) pathways. Further analysis employed gene ontology (GO) analysis: high-enrichment GOs identified endocytosis, inflammatory, apoptosis, and transport pathways. Path-Net and Signal-Net analysis highlighted the MAPK pathway as the key regulatory pathway. Moreover, the key differentially expressed genes of the significant pathways were apoptosis-related. These findings demonstrate previously unrecognized changes in gene transcription that are associated with B. melitensis infection of macrophages, and the central signaling pathways identified here merit further investigation. Our data provide new insights into the molecular attenuation mechanism of strain M5-90 and will facilitate the generation of new attenuated vaccine strains with enhanced efficacy.
Insights
Brucella melitensis strain M5-90, an attenuated vaccine, shares survival strategies with virulent strain M28 in macrophages. Key gene expression changes involve apoptosis and the MAPK pathway, offering insights into vaccine attenuation.
Area of Science:
- * Microbiology and Immunology
- * Molecular Biology and Genomics
Background:
- * Brucella melitensis causes brucellosis, a zoonotic disease impacting livestock and humans.
- * The attenuated B. melitensis strain M5-90 is a vaccine used in China, but its molecular attenuation mechanisms are unclear.
- * Understanding the host-pathogen interaction at a molecular level is crucial for vaccine development.
Purpose of the Study:
- * To investigate the genome-wide gene expression profiles of mouse macrophages in response to infection with B. melitensis strains M28 and M5-90.
- * To identify key molecular pathways and differentially expressed genes involved in the host response and vaccine attenuation.
Main Methods:
- * Genome-wide digital gene expression (DGE) analysis using the Illumina Genome Analyzer platform.
- * Analysis of differentially expressed transcripts, gene ontology (GO) analysis, Path-Net, and Signal-Net.
- * Focus on macrophage responses 4 hours post-infection.
Main Results:
- * Both M28 and M5-90 induced similar gene expression profiles in macrophages, including anti-inflammatory and antiapoptotic factors.
- * 1019 differentially expressed macrophage transcripts were identified, with significant enrichment in lysosome, MAPK, endocytosis, inflammatory, and apoptosis pathways.
- * The mitogen-activated protein kinase (MAPK) pathway was identified as a key regulatory pathway, with apoptosis-related genes being central.
Conclusions:
- * Brucella melitensis infection of macrophages triggers significant, previously unrecognized changes in gene transcription.
- * The MAPK and apoptosis pathways are central to the host response and likely involved in the attenuation of strain M5-90.
- * These findings provide insights into the molecular attenuation of B. melitensis M5-90 and can aid in developing improved vaccine strains.
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