Transcriptome Sequencing Reveals That Intact Expression of the Chicken Endogenous Retrovirus chERV3 In Vitro Can Possibly Block the Key Innate Immune Pathway.
Transcriptome Sequencing Reveals That Intact Expression of the Chicken Endogenous Retrovirus chERV3 In Vitro Can Possibly Block the Key Innate Immune Pathway.
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DOI：
10.3390/ani13172720
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发表时间：
2023-08-26
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内源性逆转录病毒（ERV）是古老的病毒序列，已整合到脊椎动物的基因组中。为了评估chERV3对宿主先天免疫反应的功能，构建了全长（10,218 bp）反向克隆质粒（puc57-chERV3）并转染到原代鸡胚成纤维细胞（CEF）中进行转录组测序。然后我们进行转录组测序来分析 chERV3 诱导的基因表达变化。我们发现chERV3下调了许多涉及免疫相关过程的基因，例如炎症反应、先天免疫反应和Toll样受体信号通路。这些过程涉及病毒感染的检测和响应。我们的研究系统地鉴定了 chERV3 原核表达对鸡先天免疫反应的影响。 ERV 是脊椎动物遗传多样性和创新的丰富来源，了解其作用和机制可能对动物疾病的易感性产生影响。内源性逆转录病毒（ERV）是已整合到脊椎动物基因组中的病毒序列。我们的初步转录组测序分析显示 chERV3 具有活性，位于染色体 1:32602284–32615631。我们假设 chERV3 可能在宿主对病毒感染的先天免疫反应中发挥作用。在本研究中，我们利用反向遗传学，在体外构建了puc57-chERV3全长反向克隆质粒。我们通过酶联免疫吸附测定（ELISA）测量了培养物上清液中p27的含量。最后，通过转录组分析来分析chERV3在先天免疫中的功能。结果表明chERV3可能产生p27病毒颗粒。我们发现，与阴性对照（NC）组（转染pMD18T-EGFP）相比，chERV3组在24小时（h）时表现出2538个上调的差异表达基因（DEG）和1828个下调的DEG，在48小时（h）时表现出1752个上调的DEG和1282个下调的DEG。基于基因本体论（GO）和京都基因与基因组百科全书（KEGG）通路富集分析，下调的DEG主要富集在炎症反应、先天免疫反应、Toll样受体信号通路等免疫相关过程中。 GSEA 显示 Toll 样受体信号通路在两个时间点均被 chERV3 抑制。我们推测chERV3可以通过阻断Toll样受体信号通路影响先天免疫通路的激活，从而实现免疫逃避。
Endogenous retroviruses (ERVs) are ancient viral sequences that have integrated into the genomes of vertebrates. In order to evaluate the function of chERV3 on the host innate immune response, a full-length (10,218 bp) reverse cloning plasmid (puc57-chERV3) was constructed and transfected into primary chicken embryo fibroblasts (CEFs) for transcriptome sequencing. We then performed transcriptome sequencing to analyze the gene expression changes induced by chERV3. We found that chERV3 down-regulated many genes involved in immune-related processes, such as the inflammatory response, innate immune response, and the Toll-like receptor signaling pathway. These processes are involved in detecting and responding to viral infection. Our study systematically identified the effect of chERV3 prokaryotic expression on the innate immune response in chickens. ERVs are a rich source of genetic diversity and innovation in vertebrates, and understanding their roles and mechanisms may have implications for susceptibility to animal diseases. Endogenous retroviruses (ERVs) are viral sequences that have integrated into the genomes of vertebrates. Our preliminary transcriptome sequencing analysis revealed that chERV3 is active and is located on chromosome 1:32602284–32615631. We hypothesized that chERV3 may have a role in the host innate immune response to viral infection. In this study, using reverse genetics, we constructed the puc57-chERV3 full-length reverse cloning plasmid in vitro. We measured the p27 content in culture supernatant by enzyme-linked immunosorbent assay (ELISA). Finally, transcriptome analysis was performed to analyze the function of chERV3 in innate immunity. The results showed that chERV3 may generate p27 viral particles. We found that compared to the negative control (NC) group (transfected with pMD18T-EGFP), the chERV3 group exhibited 2538 up-regulated differentially expressed genes (DEGs) and 1828 down-regulated DEGs at 24 hours (h) and 1752 up-regulated DEGs and 1282 down-regulated DEGs at 48 h. Based on Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses, the down-regulated DEGs were enriched mainly in immune-related processes such as the inflammatory response, innate immune response, and Toll-like receptor signaling pathway. GSEA showed that the Toll-like receptor signaling pathway was suppressed by chERV3 at both time points. We hypothesized that chERV3 can influence the activation of the innate immune pathway by blocking the Toll-like receptor signaling pathway to achieve immune evasion.
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