Project description:Cystic echinococcosis (CE) is a common zoonotic parasitic disease that seriously impacts public health. However, the full spectrum of immune cell changes in Echinococcus granulosus (E. granulosus) infection, especially the negative immune regulation of subpopulations of regulatory T cells (Treg), are not yet well understood. In this study, we aimed to identify the panoramic landscape of changes in immune cells and molecules and the distinct alterations in Treg subpopulations in mice infected with E. granulosus. We used single-cell RNA sequencing and single-cell T/B cell receptor sequencing to analyze 53,298 cells from the spleen and peripheral blood mononuclear cells (PBMCs) of healthy and E. granulosus-infected mice. We used immunofluorescence combined with RNA fluorescence in situ hybridization and quantitative real-time polymerase chain reaction to verify the sequencing results. Our results showed organ-specific immune system alterations in mice infected with E. granulosus. E. granulosus-infected mice induced a subpopulation of CD4+ cells with type I interferon production potential. Furthermore, there were seven different Treg cell subpopulations in vivo at three stages of differentiation, and Treg subpopulations of different classes and different stages of differentiation showed tissue specificity. After infection, the Lag3hi Treg and Gpr83+Igfbp4+ naïve Treg subpopulations were specifically induced in PBMCs and spleen, respectively. Furthermore, T follicular helper 2(Tfh2) cells with high expression of Cxxc5 and Spock2 were found in E. granulosus-infected mice. Additionally, combined single-cell immunome repertoire sequencing analysis revealed that gene fragment expression of multiple VJ-paired combinations of T cell receptor β chains were downregulated after infection. Our data uncovered changes in the full spectrum of immune cells in mice following E. granulosus infection, including subpopulations of cells that have not been emphasized in previous studies. These results further enrich the study of the bidirectional immunomodulatory mechanism and offer a different perspective for subsequent studies of E. granulosus infection.

Project description:The cestodes Echinococcus granulosus and Echinococcus multilocularis, as the pathogens of cystic echinococcosis and alveolar echinococcosis respectively, can cause significant health problems to the host and considerable socio-economic losses as a consequence. Based on the genomic data regarding these two species available in public database recently, we carried out high-throughput mRNA and small RNA transcriptomic sequencing of them and generate enormous transcriptomic datasets. A total of 34,717,856 reads (79.79%) mapped to E. granulosus genome, and 38,882,179 reads (87.61%) mapped to E. multiloculari genome. A total of 24,550 (7,925 known and 16,625 novel transcripts) and 23,771 transcripts (8,432 known and 15,339 novel transcripts) were assembled for E. granulosus and E. multilocularis respectively, and the assembly yielded 11,330 genes (6,815 known and 4,515 novel genes) for E. granulosus and 10,101 genes (7,051 known and 3,050 novel genes) for E. multilocularis, compared with the reference genome data. Bioinformatic analysis identified 6,826 AS events from 3,774 E. granulosus genes (33.31%) and 6,644 AS events in 3,611 E. multilocularis genes (35.75%). A total of 76,674 distinct microRNAs of E. granulosus and 115,742 of E. multilocularis were also obtained from small RNA transcriptome sequencing reads. Of these, there were 20 microRNAs of E. granulosus and 22 microRNAs of E. multilocularis that belonged to 19 and 21 microRNA families common to other metazoan lineages separately. 76 and 90 novel microRNAs so far unique to E. granulosus and E. multilocularis were also identified respectively. This study represents an extensive mRNA and small RNA transcriptome dataset obtained from the deep sequencing of these two cestode species. The findings will facilitate a more fundamental understanding of cestode biology, evolution, the host-parasite interplay, and provide new insights into the pathophysiology of echinococcosis, contributing to the development of improved interventions for disease control.

			Action	DRS
	SRR22008222_1.fastq.gz	Fastqsanger.gz
	SRR22008222_2.fastq.gz	Fastqsanger.gz
	SRR22008223_1.fastq.gz	Fastqsanger.gz
	SRR22008223_2.fastq.gz	Fastqsanger.gz
	SRR22008224_1.fastq.gz	Fastqsanger.gz

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Single-cell RNA sequencing reveals unique alterations in the immune landscape and Treg subpopulations in mice after Echinococcus granulosus infection

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