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Effect of human child or adult digestion reproduced in a dynamic in vitro gastrointestinal model on Cryptosporidium parvum transcriptome

ABSTRACT: Effect of human child or adult digestion reproduced in a dynamic in vitro gastrointestinal model on Cryptosporidium parvum transcriptome

PROVIDER: PRJNA1130464 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR29664562.fastq.gz	Fastqsanger.gz
	SRR29664563.fastq.gz	Fastqsanger.gz
	SRR29664564.fastq.gz	Fastqsanger.gz
	SRR29664565.fastq.gz	Fastqsanger.gz
	SRR29664566.fastq.gz	Fastqsanger.gz

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Effect of human child or adult digestion reproduced in a dynamic in vitro gastrointestinal model on Cryptosporidium parvum transcriptome

Project description:The impact of digestive physicochemical parameters on Cryptosporidium parvum transcriptome was analysed in a human and age-dependent context using a dynamic in vitro gastrointestinal model. The sophisticated TIM-1 in vitro model was used for a comparative analysis of C. parvum gene expression under the digestive conditions encountered in young children (from 6 months to 2 years) or in adults following the simulated ingestion of a glass of water contaminated with C. parvum oocysts. The C. parvum gene expression profile was analysed by RNA-sequencing in the inoculum, as well as the gastric and ileal effluents collected from the TIM-1 system. Results showed that time and compartment of digestion lead to the strongest modification of parasite transcriptome. Accordingly, the highest number of differentially expressed genes was observed in parasite samples collected in child or adult ileal effluents between 120 and 180 min of digestion, where invasive stages (sporozoites) are predominant.

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Project description:The Cryptosporidium parvum (C. parvum) oocyst wall provides strong protection against hostile environmental factors; however, research is limited concerning about the oocyst wall at the proteomic level. In this study, a comprehensive analysis of the proteome expressed by the oocyst wall of C. parvum was performed using label-free qualitative high-performance liquid chromatography (HPLC) fractionation and mass spectrometry-based qualitative proteomics technologies. A total of 798 proteins were identified, accounting for about 20% of the CryptoDB proteome. By using bioinformatic analysis, functional annotation and subcellular localization of the identified proteins were examined for better understanding of the characteristics of the oocyst wall. Among the identified proteins, one protein encoded by the C. parvum cgd7_5140 (Cpcgd7_5140) gene was predicted to be located on the surface of the oocyst wall. To verify its localization, an indirect immunofluorescent antibody assay (IFA) demonstrated that the Cpcgd7_5140 was localized on the surface of the oocyst wall, illustrating the potential usage as a marker for C. parvum detection in vitro. The results provide new information about the proteomic composition of the Cryptosporidium oocyst wall, thereby providing a theoretical basis for further study of Cryptosporidium oocyst wall formation as well as the selection of targets for Cryptosporidium detection.

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