基于转录组测序分析细粒棘球绦虫原头蚴与巨噬细胞RAW264.7免疫互作相关基因

doi:10.11843/j.issn.0366-6964.2022.11.023

Abstract

Abstract: In this study, we report the analysis of the genes involved in the immune interaction between Echinococcus granulosus protoscoleces and macrophage RAW264.7, which provides a theoretical basis for further elucidating the mechanism of immune regulation and parasitic adaptation of the protoscoleces. The protoscolices and macrophage RAW264.7 cells were co-cultured for 24 hours, then the protoscolices were collected, total RNA was extracted and cDNA library was constructed. The results showed that there were 435 genes significantly differentially expressed in the protoscolices after co-cultured with macrophages RAW264.7 for 24 h compared with 0 h, which included 227 up-regulated genes, such as HSP70, HSP10, EG95 precursor, AgB, FABP and vesicular transporter SC22B; And 208 down-regulated genes, such as EF-hand, Cathepsin, transmembrane protein 144, endosterol receptor and MAPK7. GO analysis showed that the differentially expressed genes (DEGs) were mainly involved in heme transport, iron coordinated entity transport, extracellular Matrix, Ribonuclease MRP complex, serine type endopeptidase inhibitor activity and Galactosidases activity. KEGG analysis showed that the DEGs were mainly related to spliceosome, endocytosis, endoplasmic reticulum protein processing, phagosomes, MAPK signaling and calcium signaling. When macrophages RAW264.7 were co-cultured with protoscolex for 24 hours, compared with PBS control group, the expression of 3 745 genes in RAW264.7 cells was significantly changed, of which 1 159 genes were up-regulated, 2 586 genes down-regulated. The results of GO analysis showed that the DEGs were mainly concentrated in metabolic processes, intracellular components, intracellular, Organelles and membrane-structured Organelles. The analysis of KEGG showed that the DEGs were mainly involved in metabolic pathway, ribosome pathway, spliceosome pathway, RNA transport and ubiquitin-mediated protein hydrolysis. At the same time, some DEGs were selected randomly and verified by qRT-PCR. The results showed that the expression trend was consistent with that of RNA-seq results, which further indicates that the transcriptome data is credible. In conclusion, when the protoscoleces are subjected to immunologic pressure from host macrophages, they may cause gene differential expression, the immunoregulatory molecules such as AgB, FABP1 and Kunitz-type Serine protease inhibitors in protoscoleces were up-regulated, which might be involved in host immunoregulation, moreover, it is beneficial to parasitism and immune escape for E.granulosus.

Key words: Echinococcus granulosus, macrophages RAW264.7, differential expression, immune regulation

CLC Number:

S852.734

WANG Zhengrong, MA Xun, ZHANG Yanyan, MENG Jimeng, BO Xinwen. Analysis of Genes Related to Immune Interaction between Protoscolices of Echinococcus granulosus and Macrophage RAW264.7 by Transcriptome Sequencing[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(11): 3975-3988.

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Analysis of Genes Related to Immune Interaction between Protoscolices of Echinococcus granulosus and Macrophage RAW264.7 by Transcriptome Sequencing

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