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Automated Separation of C. elegans Variably Colonized by a Bacterial Pathogen
Published on: March 21, 2014
Subsequent infection differentially affects the proteome of Caenorhabditis elegans by abrogating the intestinal cell
Prithika Udayakumar1, Balamurugan Krishnaswamy1
1Department of Biotechnology, Science Campus, Alagappa University, Karaikudi, 630 003, Tamil Nadu, India.
Abstract:
With a wide range of bacterial infections growing, it has become a big challenge to the research field to combat the newly emerging diseases. Immuno-compromised patients are vulnerable to opportunistic infections. P. mirabilis, an opportunistic pathogen infects the nematode when the immune system is compromised. In the present study, the C. elegans was pre-exposed to S. aureus for a short term, and then consecutively infected with P. mirabilis. The primary infection caused by S. aureus makes the immune system of C. elegans vulnerable making it easy for P. mirabilis to colonize efficiently during subsequent exposure, thereby stimulating the immune system of the nematode. In this study, the C. elegans exposed to the pathogens (S. aureus 4 h/P. mirabilis 40 h and S. aureus 8 h/P. mirabilis 60 h time points) showed a substantial difference in the banding patterns of SDS-PAGE gel, when compared to their respective OP50 fed controls. 2-DE identified a total of 235 proteins from all the time points which had >2 fold regulation. The regulated protein spots were identified by MALDI-ToF/ToF analysis and one common protein CDC-25.1 was found to be regulated in all the comparative time points. CDC-25.1 seemed to down regulate during subsequent infection and up regulate in single infection. The transcriptomic regulation of cdc-25.1 also reflects the protein regulation. In addition to it, survival assay in cdc-25.1 mutant nematodes confirm the susceptibility of host during subsequent infection.
Insights
Primary infection with Staphylococcus aureus compromises the nematode
Area of Science:
- Microbiology and Immunology
- Nematode Pathogenesis Research
Background:
- Emerging bacterial infections pose significant challenges, particularly for immunocompromised individuals susceptible to opportunistic pathogens.
- Proteus mirabilis is an opportunistic pathogen that infects nematodes like Caenorhabditis elegans when their immune systems are weakened.
Purpose of the Study:
- To investigate the impact of sequential bacterial infections on the immune response of Caenorhabditis elegans.
- To identify host proteins regulated during opportunistic infections and their role in host susceptibility.
Main Methods:
- Caenorhabditis elegans were sequentially infected with Staphylococcus aureus followed by Proteus mirabilis.
- Proteomic analysis using SDS-PAGE and 2-DE was performed to identify differentially expressed proteins.
- MALDI-ToF/ToF was used for protein identification, and transcriptomic analysis and survival assays were conducted.
Main Results:
- Sequential infection with S. aureus and P. mirabilis significantly altered protein expression patterns in C. elegans compared to controls.
- 235 proteins showed >2 fold regulation, with CDC-25.1 identified as a key regulated protein.
- CDC-25.1 was downregulated during sequential infection and upregulated during single infection, with its transcriptomic regulation mirroring protein levels.
Conclusions:
- Sequential bacterial infections can modulate the host immune response in C. elegans, increasing susceptibility.
- The protein CDC-25.1 plays a role in host defense against opportunistic pathogens, with its regulation critical during sequential infections.
