目的:通过对EBV刺激过表达CD21 的Jurkat T细胞进行转录组测序分析，研究EBV感染T细胞的机制。方法:经过PCR技术克隆目的基因CD21，再通过分子克隆技术将 CD21 基因插入到过表达慢病毒载体上，使其在Jurkat T细胞表面稳定表达。通过EBV刺激过表达CD21 的Jurkat T细胞(EBV-LV-Jurkat)，然后与未经处理的Jurkat T细胞和仅经过慢病毒感染的 Jurkat T 细胞(LV-Jurkat)均进行普通转录组测序，并对EBV-LV-Jurkat T 与LV-Jurkat T这两组进行差异表达基因的富集通路分析。结果:通过对EBV感染过表达CD21 的Jurkat T细胞(EBV-LV-Jurkat T)与过表达 CD21 的 Jurkat T细胞(LV-Jurkat T)差异表达基因(DEGs)GO 富集分析发现，其中在分子功能方面主要富集于 MAP激酶酪氨酸磷酸酶活性信号通路，其相关基因DUSP1、DUSP10、DUSP5、DUSP8 等被显著富集。对 EBV-LV-Jurkat T与LV-Jurkat T的DEGs进行KEGG 代谢通路分析发现，多种与 EBV 感染 B 细胞之后相似的细胞通路以及分子主要富集于MAPK、IL-17、HIF-1、NF-κB 与 PI3K/AKT 等信号通路，以及免疫反应系统相关基因也被显著富集。结论:成功构建了CD21 过表达的Jurkat T 细胞模型，并通过转录组测序揭示了EBV刺激能够显著上调MAPK信号通路。这为进一步探索EBV感染T细胞的机制提供了重要的初步数据。
Physiological and Biochemical Characteristics and Transcriptome Analysis of EBV Infected Jurkat T Cells
Objective:This study aims to investigate the mechanism of EBV infection in T cells by se-quencing the transcriptome of EBV-stimulated Jurkat T cells overexpressing CD21.Methods:The target gene CD21 was cloned by PCR and inserted into an overexpression lentiviral vector via molecular cloning tech-niques,ensuring its stable overexpression on the surface of Jurkat T cells.The CD21-overexpressing Jurkat T cells were then stimulated with EBV(EBV-LV-Jurkat).Transcriptome sequencing was performed on EBV-stimulated CD21-overexpressing Jurkat T cells,untreated Jurkat T cells,and Jurkat T cells infected with lentivirus alone(LV-Jurkat).Gene set enrichment analysis was conducted on differentially expressed genes(DEGs)between EBV-stimulated CD21-overexpressing Jurkat T cells(EBV-LV-Jurkat T)and CD21-overexpressing Jurkat T cells(LV-Jurkat T).Results:GO enrichment analysis of DEGs between EBV-LV-Jurkat T and LV-Jurkat T revealed significant enrichment in the molecular function of MAP kinase tyrosine phosphatase activity,involving the DUSP1,DUSP10,DUSP5,and DUSP8 genes.KEGG pathway enrich-ment analysis of DEGs indicated that multiple pathways and molecules,similar to those observed in EBV-in-fected B cells,were significantly enriched in the MAPK,IL-17,HIF-1,NF-κB,and PI3K/AKT signaling pathways,and genes related to the immune response system.Conclusion:This study successfully constructed a model of CD21-overexpressing Jurkat T cells and revealed the impact of EBV stimulation on significantly ac-tivating the MAPK signaling pathway,providing crucial preliminary data for further exploration of the mecha-nism of EBV infection in T cells.
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