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在原代和连续细胞培养物中,用野生型和糖蛋白 g 缺失(ΔgG)ILTV 株体外感染时的宿主-病毒相互作用的转录组分析。

Transcriptomic analyses of host-virus interactions during in vitro infection with wild-type and glycoprotein g-deficient (ΔgG) strains of ILTV in primary and continuous cell cultures.

机构信息

Faculty of Science, Asia-Pacific Centre for Animal Health, Melbourne Veterinary School, The University of Melbourne, Melbourne, Victoria, Australia.

Escuela de Medicina Veterinaria, Universidad Andrés Bello, Concepción, Biobío, Chile.

出版信息

PLoS One. 2024 Oct 11;19(10):e0311874. doi: 10.1371/journal.pone.0311874. eCollection 2024.

Abstract

Infectious laryngotracheitis (ILT) remains a significant concern for the poultry industry worldwide due to its impact on animal welfare and its substantial economic consequences. The disease is caused by the alphaherpesvirus, infectious laryngotracheitis virus (ILTV). This study investigated in vitro host-virus interactions of a glycoprotein G (gG) deletion mutant vaccine strain of ILTV (ΔgG ILTV), and its parent wild-type strain (CSW-1 ILTV). Inoculations were performed separately for the two strains of ILTV using both a primary (chicken embryonic kidney, CEK) and a continuous culture (leghorn male hepatoma, LMH) of chicken cells. Transcriptome analysis was performed at 12 hours post infection. Each cell-type displayed distinct effects on host and viral gene transcription, with a greater number of viral and host genes differentially transcribed in CEK cells and LMH cells, respectively. Both cell-types infected with either strain demonstrated enrichment of pathways related to signalling, and gene ontologies (GO) associated with chemotaxis. Infection with either strain upregulated both SOCS proteins and certain proto-oncogenes, which may contribute to prolonged viral persistence by promoting immunosuppression and preventing apoptosis, respectively. Patterns of gene transcription related to cytokines, chemokines, endosomal TLRs, and interferon responses, as well as pathways associated with histone acetylation, transport, and extracellular matrix organization were similar within each cell type, regardless of the viral strain. In CEK cells, GO terms and pathways were downregulated uniquely after CSW-1 ILTV infection, indicating a viral-strain specific effect in this cell-type. Overall, this study highlights that the observed differences in host and ILTV gene transcription in vitro were more strongly influenced by the cell-types used rather than the presence or absence of gG. This underscores the importance of cell-line selection in studying host-virus interactions and interpreting experimental results.

摘要

传染性喉气管炎(ILT)仍然是全球家禽业的一个重大关注点,因为它会影响动物福利并造成巨大的经济后果。该疾病由α疱疹病毒、传染性喉气管炎病毒(ILTV)引起。本研究调查了 ILTV 的糖蛋白 G(gG)缺失突变疫苗株(ΔgG ILTV)及其亲本野生型株(CSW-1 ILTV)的体外宿主-病毒相互作用。分别使用原代(鸡胚肾,CEK)和鸡连续培养(来亨雄性肝细胞,LMH)对两种 ILTV 株进行接种。在感染后 12 小时进行转录组分析。两种细胞类型对宿主和病毒基因转录都表现出不同的影响,CEK 细胞和 LMH 细胞分别有更多的病毒和宿主基因差异转录。两种细胞类型感染两种毒株均表现出与信号转导相关的途径和与趋化作用相关的基因本体(GO)富集。感染任一毒株均可上调 SOCS 蛋白和某些原癌基因的表达,这可能分别通过促进免疫抑制和防止细胞凋亡来促进病毒持续存在。细胞内无论病毒株如何,与细胞因子、趋化因子、内体 TLR 和干扰素反应相关的基因转录模式以及与组蛋白乙酰化、运输和细胞外基质组织相关的途径相似。在 CEK 细胞中,CSW-1 ILTV 感染后特异地下调了 GO 术语和途径,表明该细胞类型中存在病毒株特异性效应。总体而言,本研究强调了体外观察到的宿主和 ILTV 基因转录差异更多地受到所用细胞类型的影响,而不是 gG 的存在与否。这突出了在研究宿主-病毒相互作用和解释实验结果时细胞系选择的重要性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/821e/11469545/26019b812b96/pone.0311874.g001.jpg

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