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Proteomic Analysis of Fasciola hepatica Excretory and Secretory Products Co-Immunoprecipitated Using Time Course Infection Sera.

ABSTRACT: Fasciola hepatica is a widespread pathogen that is known for its harmful effects on the health and productivity of ruminant animals. To identify the proteins present in all periods of infection with F. hepatica but not in those with Fasciola gigantica by shotgun liquid chromatography-tandem mass spectrometry (LC-MS/MS), we collected the ESPs and sera of F. hepatica and F. gigantica. In this study, the sheep were artificially infected with F. hepatica and the sera were collected at five different periods: 3 days post-infection (dpi), 7 dpi, 21 dpi, 63 dpi, and 112 dpi. The interacting proteins were pulled down from the sheep sera of all five periods and the sera with F. gigantica by co-immunoprecipitation (Co-IP) assay, before being identified by LC-MS/MS analysis. Thirty, twenty-two, twenty-three, twenty-seven, and twenty-two proteins were pulled down by the infected sera at 3 dpi, 7 dpi, 21 dpi, 63 dpi, and 112 dpi, respectively. Among them, 12 proteins existed in all periods, while six proteins could be detected in all periods in F. hepatica but not in F. gigantica. Protein relative pathway analysis revealed that these proteins mainly refer to the metabolism, regulation of genetic activity, and signal transduction of F. hepatica. In conclusion, this study provides meaningful data for the diagnosis of fasciolosis and to understand the interactions between F. hepatica and the host.

SUBMITTER: Lan Z

PROVIDER: S-EPMC8231988 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Widespread Fasciola gigantica infection in buffaloes has caused great economic losses in buffalo farming. Studies on F. gigantica excretory and secretory products (FgESP) have highlighted their importance in F. gigantica parasitism and their potential in vaccine development. Identifying FgESP components involved in F. gigantica-buffalo interactions during different periods is important for developing effective strategies against fasciolosis. Buffaloes were assigned to non-infection (n = 3, as control group) and infection (n = 3) groups. The infection group was orally administrated 250 metacercariae. Sera were collected at 3, 10, and 16 weeks post-infection (wpi) for the non-infection group and at 0 (pre-infection), 1, 3, 6, 8, 10, 13, and 16 wpi for the infection group. FgESP components interacting with sera from the non-infection and infection groups assay were pulled down by co-IP and identified using LC-MS/MS. Interacting FgESP components in infection group were subjected to Kyoto Encyclopedia of Genes and Genomes (KEGG) metabolic pathway and gene ontology (GO) functional annotation to infer their potential functions. Proteins of FgESP components identified in the non-infection group at 3, 10, and 16 wpi accounted for 80.5%, 84.3%, and 82.1% of all proteins identified in these 3 time points, respectively, indicating surroundings did not affect buffalo immune response during maintenance. 490 proteins were identified in the infection group, of which 87 were consistently identified at 7 time points. Following GO analysis showed that most of these 87 proteins were in biological processes, while KEGG analysis showed they mainly functioned in metabolism and cellular processing, some of which were thought to functions throughout the infection process. The numbers of specific interactors identified for each week were 1 (n = 12), 3 (n = 5), 6 (n = 8), 8 (n = 15), 10 (n = 23), 13 (n = 22), and 16 (n = 14) wpi, some of which were thought to functions in specific infection precess. This study screened the antigenic targets in FgESP during a dense time course over a long period. These findings may enhance the understanding of molecular F. gigantica-buffalo interactions and help identify new potential vaccine and drug target candidates.

Project description:Introduction: The secondary infection of F. gigantica 4 weeks after primary infection could not induce a robust adaptive immune response in buffalo. As excretory and secretory products (ESP) show intense immune regulation and plays critical role in primary infection process, while research on ESP in secondary infection is lacking. Does the components of ESP in secondary infection was different from that of primary infection? Does fluke secrete and excrete new component to immunoregulation during secondary infection? Here, the component of F. gigantica ESP in buffalo primary and secondary infections was identified and compared, which will drive the exploration in repeat infection in basis matter, and also conducive to drug targets and vaccine candidate screening. Methods: Buffaloes were assigned to the group A (n = 3, non-infection), group B (n = 3, primary infection) and group C (n = 3, secondary infection). The primary infection group was orally administrated 250 metacercariae at 4 week post-infection (wpi), while the secondary infection group was orally administrated 250 metacercariae at 0 and 4 wpi. Then Sera were collected at 0, 10, and 16 wpi for group A, 0 (pre-infection), 1, 3, 6, 8, 10, 13, and 16 wpi for group B, and 0, 1, 3, 4, 5, 7, 10, 12, 14, 17 and 20 wpi for group C. FgESP components interacting with sera from three groups were pulled down by co-immunoprecipitated (co-IP) and identified using LC–MS/MS. Interacting FgESP components in group C were subjected to Kyoto Encyclopedia of Genes and Genomes (KEGG) metabolic pathway and gene ontology (GO) functional annotation to infer their potential functions, and also been compared with that of group B. Results and discussion: In the group C, 327 proteins were identified, of which 74 were consistently identified at 7 time points (5, 7, 10, 12, 14, 17 and 20 wpi). GO analysis shows these 74 proteins mainly functioned in biological processes, while KEGG analysis showed they mainly functioned in metabolism and cellular processing. The numbers of specific interactors identified were 2 (5 wpi), 10 (7 wpi), 0 (10 wpi), 8 (12 wpi), 1 (14 wpi), 4 (17 wpi), and 4 (20 wpi). Here, the antigenic targets in FgESP in the secondary infection process were screened over a long and intensive period of time, and these components needs further exploration. The present study will deepen the understanding of molecular F. gigantica-buffalo interactions and helpful in new potential vaccine and drug target candidates identification

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Proteomic Analysis of Fasciola hepatica Excretory and Secretory Products Co-Immunoprecipitated Using Time Course Infection Sera.

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