ABSTRACT
OBJECTIVE@#To investigate the serum microRNA (miRNA) expression and examine the impact of miRNA expression profiles on T helper type 17 (Th17)/regulatory T cells (Treg) imbalance among patients with cystic echinococcosis, so as to provide insights into the illustration of the mechanisms underlying chronic Echinococcus granulosus infections, and long-term pathogenesis.@*METHODS@#Total RNA was extracted from the sera of cystic echinococcosis patients and healthy controls, and subjected to high-throughput sequencing with the Illumina sequencing platform. Known miRNAs were annotated and new miRNAs were predicted using the miRBase database and the miRDeep2 tool, and differentially expressed miRNAs were identified. The target genes of differentially expressed miRNAs were predicted using the software miRanda and TargetScan, and the intersection was selected for Gene Ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. Among the differentially expressed miRNAs with the 20 highest fold changes, miRNAs that targeted genes relating to key transcription factors RORC and FOXP3 that determine the production of Th17 and Treg cells or their important regulatory pathways (PI3K-Akt and mTOR pathways) were matched.@*RESULTS@#A total of 53 differentially expressed miRNAs were screened in sera of cystic echinococcosis patients and healthy controls, including 47 up-regulated miRNAs and 6 down-regulated miRNAs. GO enrichment analysis showed that these differentially expressed miRNA were involved DNA transcription and translation, cell components, cell morphology, neurodevelopment and metabolic decomposition, and KEGG pathway analysis showed that the differentially expressed miRNA were mainly involved in MAPK, PI3K-Akt and mTOR signaling pathways. Among the differentially expressed miRNAs with the 20 highest fold changes, there were 3 miRNAs that had a potential for target regulation of RORC, and 15 miRNAs that had a potential to target the PI3K-Akt and mTOR signaling pathways.@*CONCLUSIONS@#Significant changes are found in serum miRNA expression profiles among patients with E. granulosus infections, and differentially expressed miRNAs may lead to Th17/Treg imbalance through targeting the key transcription factors of Th17/Treg or PI3K-Akt and mTOR pathways, which facilitates the long-term parasitism of E. granulosus in hosts and causes a chronic disease.
Subject(s)
Humans , Echinococcosis/genetics , Gene Expression Profiling , MicroRNAs/metabolism , Phosphatidylinositol 3-Kinases/genetics , Proto-Oncogene Proteins c-akt/genetics , T-Lymphocytes, Regulatory , TOR Serine-Threonine Kinases/genetics , Th17 Cells , Transcription Factors/geneticsABSTRACT
We screened echinococcosis-specific diagnostic antigen and verified its immunogenicity.The sequencing data of Echinococcus granulosus released by the National Human Genome Research Center was analyzed.The bioinformaties method was used to screen out Eg-07279 antigen gene Eg-07279,which was not expressed in Echinococcus granulosus and highly expressed in the original metacercariae.After expression,the recombinant protein rEg-07279 was purified by affinity chromatography.The specific IgG level was detected in the recombinant protein immunized mice and the immunogenicity was verified by Western blot.The selected antigen molecule Eg-07279 was cloned,expressed and purified to obtain the recombinant protein.The results of ELISA showed that the specific IgG level of Eg-07279 (2.559±0.125) was significantly higher than that of the blank group (0.319 0±0.01),the difference was statistically significant (P＜0.01);the Western blot results showed that the recombinant plasmids could be recognized by the secondary infection of the protoscolex and the serum of the recombinant protein immunized group,while the serum of the blank control group was not recognized by the serum.In summary,the Eg07279 antigen of E.granulosus was obtained and it was proved that the recombinant protein has good immunogenicity and is a good diagnostic antigen candidate molecule.