Wani Sajad Ahmad, Praharaj Manas Ranjan, Sahu Amit R, Khan Raja Ishaq Nabi, Saxena Shikha, Rajak Kaushal Kishor, Muthuchelvan Dhanavelu, Sahoo Aditya, Mishra Bina, Singh Raj Kumar, Mishra Bishnu Prasad, Gandham Ravi Kumar
ICAR-Indian Veterinary Research Institute (IVRI), Bareilly, India.
The Ohio State University, Columbus, Ohio, USA.
mSystems. 2021 Mar 30;6(2):e00820-20. doi: 10.1128/mSystems.00820-20.
Immune response is a highly coordinated cascade involving all the subsets of peripheral blood mononuclear cells (PBMCs). In this study, RNA sequencing (RNA-Seq) analysis of PBMC subsets was done to delineate the systems biology behind immune protection of the vaccine in sheep and goats. The PBMC subsets studied were CD4, CD8, CD14, CD21, and CD335 cells from day 0 and day 5 of sheep and goats vaccinated with Sungri/96 peste des petits ruminants virus. Assessment of the immune response processes enriched by the differentially expressed genes (DEGs) in all the subsets suggested a strong dysregulation toward the development of early inflammatory microenvironment, which is very much required for differentiation of monocytes to macrophages, and activation as well as the migration of dendritic cells into the draining lymph nodes. The protein-protein interaction networks among the antiviral molecules (IFIT3, ISG15, MX1, MX2, RSAD2, ISG20, IFIT5, and IFIT1) and common DEGs across PBMC subsets in both species identified ISG15 to be a ubiquitous hub that helps in orchestrating antiviral host response against peste des petits ruminants virus (PPRV). IRF7 was found to be the key master regulator activated in most of the subsets in sheep and goats. Most of the pathways were found to be inactivated in B lymphocytes of both the species, indicating that 5 days postvaccination (dpv) is too early a time point for the B lymphocytes to react. The cell-mediated immune response and humoral immune response pathways were found more enriched in goats than in sheep. Although animals from both species survived the challenge, a contrast in pathway activation was observed in CD335 cells. Peste des petits ruminants (PPR) by PPR virus (PPRV) is an World Organisation for Animal Health (OIE)-listed acute, contagious transboundary viral disease of small ruminants. The attenuated Sungri/96 PPRV vaccine used all over India against this PPR provides long-lasting robust innate and adaptive immune response. The early antiviral response was found mediated through type I interferon-independent interferon-stimulated gene (ISG) expression. However, systems biology behind this immune response is unknown. In this study, transcriptome profiling of PBMC subsets (CD4, CD8, CD14, CD21, and CD335) in vaccinated goats and sheep (at 5 days postvaccination) was done to understand this systems biology. Though there are a few differences in the systems biology across cells (specially the NK cells) between sheep and goats, the coordinated response that is inclusive of all the cell subsets was found to be toward the induction of a strong innate immune response, which is needed for an appropriate adaptive immune response.
免疫反应是一个高度协调的级联反应，涉及外周血单个核细胞（PBMC）的所有亚群。在本研究中，对PBMC亚群进行了RNA测序（RNA-Seq）分析，以阐明绵羊和山羊疫苗免疫保护背后的系统生物学。所研究的PBMC亚群是接种Sungri/96小反刍兽疫病毒的绵羊和山羊在第0天和第5天的CD4、CD8、CD14、CD21和CD335细胞。对所有亚群中差异表达基因（DEG）富集的免疫反应过程的评估表明，早期炎症微环境的发展存在强烈的失调，而单核细胞分化为巨噬细胞以及树突状细胞激活并迁移至引流淋巴结非常需要这种微环境。两种物种PBMC亚群中的抗病毒分子（IFIT3、ISG15、MX1、MX2、RSAD2、ISG20、IFIT5和IFIT1）之间的蛋白质-蛋白质相互作用网络以及共同的DEG确定ISG15是一个普遍存在的枢纽，有助于协调针对小反刍兽疫病毒（PPRV）的抗病毒宿主反应。发现IRF7是在绵羊和山羊的大多数亚群中被激活的关键主调节因子。发现大多数通路在两种物种的B淋巴细胞中均失活，这表明接种疫苗后5天（dpv）对于B淋巴细胞反应来说时间点太早。发现山羊的细胞介导免疫反应和体液免疫反应通路比绵羊更富集。尽管两种物种的动物在攻毒后均存活，但在CD335细胞中观察到通路激活存在差异。小反刍兽疫病毒（PPRV）引起的小反刍兽疫（PPR）是世界动物卫生组织（OIE）列出的小反刍动物的一种急性、传染性跨界病毒性疾病。在印度各地用于预防PPR的减毒Sungri/96 PPRV疫苗可提供持久而强大的先天和适应性免疫反应。发现早期抗病毒反应是通过I型干扰素非依赖性干扰素刺激基因（ISG）表达介导的。然而，这种免疫反应背后的系统生物学尚不清楚。在本研究中，对接种疫苗的山羊和绵羊（接种后5天）的PBMC亚群（CD4、CD8、CD14、CD21和CD335）进行了转录组分析，以了解这种系统生物学。尽管绵羊和山羊的细胞（特别是NK细胞）之间的系统生物学存在一些差异，但发现包括所有细胞亚群的协调反应是朝着诱导强烈的先天免疫反应进行的，而这是适当的适应性免疫反应所必需的。
