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Label-Free Quantitative Proteomics Workflow for Discovery-Driven Host-Pathogen Interactions
Published on: October 20, 2020
Comparative proteomics reveals Cryptosporidium parvum manipulation of the host cell molecular expression and immune
Teng Li1,2, Hua Liu1, Nan Jiang1
1National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention (Chinese Center for Tropical Diseases Research); Key Laboratory of Parasite and Vector Biology, National Health Commission of People's Republic of China; WHO Collaborating Center for Tropical Diseases, Shanghai, China.
Abstract:
Cryptosporidium is a life-threating protozoan parasite belonging to the phylum Apicomplexa, which mainly causes gastroenteritis in a variety of vertebrate hosts. Currently, there is a re-emergence of Cryptosporidium infection; however, no fully effective drug or vaccine is available to treat Cryptosporidiosis. In the present study, to better understand the detailed interaction between the host and Cryptosporidium parvum, a large-scale label-free proteomics study was conducted to characterize the changes to the proteome induced by C. parvum infection. Among 4406 proteins identified, 121 proteins were identified as differentially abundant (> 1.5-fold cutoff, P < 0.05) in C. parvum infected HCT-8 cells compared with uninfected cells. Among them, 67 proteins were upregulated, and 54 proteins were downregulated at 36 h post infection. Analysis of the differentially abundant proteins revealed an interferon-centered immune response of the host cells against C. parvum infection and extensive inhibition of metabolism-related enzymes in the host cells caused by infection. Several proteins were further verified using quantitative real-time reverse transcription polymerase chain reaction and western blotting. This systematic analysis of the proteomics of C. parvum-infected HCT-8 cells identified a wide range of functional proteins that participate in host anti-parasite immunity or act as potential targets during infection, providing new insights into the molecular mechanism of C. parvum infection.
Insights
Cryptosporidium parvum infection triggers host immune responses and inhibits host cell metabolism. This proteomics study reveals key proteins involved in host immunity and potential therapeutic targets for cryptosporidiosis.
Area of Science:
- Proteomics
- Host-pathogen interactions
- Immunology
Background:
- Cryptosporidium is a protozoan parasite causing gastroenteritis.
- Cryptosporidiosis is re-emerging, with no fully effective treatments.
- Understanding host-parasite interactions is crucial for developing interventions.
Purpose of the Study:
- To characterize proteome changes in host cells infected with Cryptosporidium parvum.
- To identify host proteins involved in anti-parasite immunity.
- To uncover potential therapeutic targets for cryptosporidiosis.
Main Methods:
- Label-free proteomics analysis of C. parvum-infected HCT-8 cells.
- Quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR).
- Western blotting for protein validation.
Main Results:
- 121 differentially abundant proteins identified in infected cells (67 upregulated, 54 downregulated).
- Host cells mount an interferon-centered immune response against C. parvum.
- C. parvum infection extensively inhibits host cell metabolism-related enzymes.
Conclusions:
- Proteomics analysis provides insights into the molecular mechanisms of C. parvum infection.
- Identified proteins are involved in host immunity or represent potential therapeutic targets.
- Further research into these proteins may lead to new treatments for cryptosporidiosis.
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