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:Purporse: FPV can cause a viral disease that occurs in cats, causing severe leukopenia, gastro-enteritis and nervous signs and leads to significant losses. LncRNAs biochemically resemble mRNAs posited by Jacob and Monod, yet do not template protein synthesis. Rather, lncRNAs functioning as RNA genes to play an important role in many biological and pathological processes such as inflammation and viral infection. However, lncRNAs have not been comprehensively identified in felines. Here, microarrays of mRNAs and lncRNAs were globally performed to identify the lncRNAs related to immune response of feline panleukopenia virus infection in F81 cells. Methods:lncRNA and mRNA sequencing analysis was performed on an Illumina Hiseq 4000 (LC Sciences, USA) following the vendor's recommended protocol Results: Through RNA sequencing, we performed a comprehensive analysis of lncRNAs and mRNAs in FPV infected F81 cells. F81 cells infected or uninfected FPV were used to prepare six libraries to generate 14.66 to 15.45 gigabytes (GB) clean reads per sample. We obtained 90588 novel lncRNAs, and 291 are differential expressed lncRNAs between FPV infected cells and uninfected cells. Consistent with previous studies, lncRNAs showed features distinct from mRNAs, such as shorter lengths, lesser exon numbers, and lower expression levels compared to mRNAs. The GO enrichment and KEGG pathway analyses showed that a majority of genes and lncRNAs were mainly related to the immune response of host cells with FPV infection. By differentially expressed target genes as well as co-expression network construction analysis, 21 key differentially expressed genes (19 upregulated & 2 downregulated) related to immune regulation of FPV infection in F81 cells were identified. Moreover, fifteen differentially expressed genes (five lncRNAs, ten mRNAs) were selected and independently validated by reverse-transcription qPCR and the results were consistent with the sequencing results. Conclusion: The present study is the first to provide insights into the biological connection of lncRNAs and FPV infection, which can be further explore for the potential link between lncRNA expression and the pathogenesis of FPV infection.

Project description:Local hypoxia is recently reported to be occur in white adipose tissue (WAT) microenvironment at obesity. Adipocyte have own life cycle that reflect in different stages of adipogenesis in WAT niche. Long noncoding RNAs (lncRNAs) play an important role in cellular response to hypoxia. However, the different hypoxic responses of preadipocytes during adipogenesis and the potential role of lncRNAs in this process remain to be elucidated. Here we evaluated the different hypoxic responses of primary hamster preadipocytes during adipogenesis and analyzed the expression of mRNAs and lncRNAs from same Ribo-Zero RNA-seq libraries. Hypoxia induced HIF-1α protein during adipogenesis and caused the divergent changes of cell phenotypes. A total of 10,318 mRNAs were identified to be expressed in twenty libraries (five timepoints), and 149, 326, 365, 1099, 2504 differentially expressed mRNAs (DE mRNAs) were detected in D0, D1, D3, D7 and D11, respectively (hypoxia vs normoxia). Functional enrichment analysis revealed the shared and specific hypoxia response pathways in different stages of adipogenesis. Hypoxia differentially modulates the expression profile of adipose-associated genes, including adipokines, lipogenesis, lipolysis, hyperplasia, hypertrophy, inflammatory and extracellular matrix (ECM). We also identified 4,296 lncRNAs that were substantially expressed and detected 1,431 DE lncRNAs in five timepoints. Afterward, 2, 3, 5, 13 and 50 DE mRNAs in D0, D1, D3, D7 and D11, respectively, were highly correlated and loci-nearby with DE lncRNAs, and were mainly involved in cell cycle, vesicle-mediated transport and mitochondrion organization. We also identified twenty-eight one-to-one lncRNA-mRNA pairs that might be closely related to adipocyte function, such as ENSCGRT00015041780-Hilpda, TU2105-Cdsn, TU17588-Ltbp3. These lncRNAs may represent the crucial regulation axis in cellular response to hypoxia during adipogenesis. Altogether, this study dissected the effects of hypoxia in the cell during adipogenesis, and uncovered novel regulator potentially associated with obesity and may provide a new view for interpretation and treatment of obesity.

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Microarray analysis of lncRNA and mRNA reveals lipogenesis inhibition along with metabolic remodeling in mice infected by the larval Echinococcus granulosus

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