Single-Cell Transcriptome Profiling of Scale Drop Disease Virus-Infected Asian Seabass (Lates calcarifer)

Zhixuan Loh, Ting Wei Lim, S. Howland, Sunita Awate, Laurent Rénia, Jinmiao Chen, Ee Chee Ren
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Abstract

The study aims to characterize the immune cell landscape in convalescent Asian seabass (Lates calcarifer) blood samples after exposure to scale-drop disease virus (SDDV). Traditional immunophenotyping approaches used in human and mouse studies are impractical for non-model organisms like the Asian seabass due to the lack of specific antibody-based reagents. To overcome this challenge, 10x Genomics single-cell RNA sequencing was employed. The analysis of blood samples revealed 24 distinct leukocyte clusters, with elevated proportions of B cells, granulocytes, and T cells in the convalescent group compared to the uninfected group. While distinguishing granulocyte and macrophage subsets was challenging, the analysis of differential gene expression in the macrophage population indicated that the upregulated genes were linked to inflammatory processes. Specific T cell clusters showed notable expressions of cd4-1, cd8a, perforin-1 and il-2rβ, suggesting the presence of CD4+ T helper (Th), CD8+ cytotoxic T (Tc) cells, immature T cells, and naive T cells. Attempts to categorize CD4+ T cells into Th subtypes lacked clear distinctions, while CD8+ T cells exhibited three clusters, predominantly Tc1 cells. Furthermore, comparisons between convalescent and uninfected groups revealed increased percentages of activated and antibody-secreting B cells in the convalescent group. This single-cell analysis provides vital insights into the immune cell dynamics in convalescent and uninfected Asian seabass, providing valuable information on potential immune responses to SDDV infection.
受鳞屑病病毒感染的亚洲鲈鱼(Lates calcarifer)的单细胞转录组分析
该研究旨在描述亚洲鲈鱼(Lates calcarifer)暴露于鳞屑病病毒(SDDV)后的康复期血液样本中免疫细胞的特征。由于缺乏特异性抗体试剂,用于人类和小鼠研究的传统免疫分型方法对于亚洲鲈鱼等非模式生物来说并不实用。为了克服这一难题,我们采用了 10x Genomics 单细胞 RNA 测序技术。对血液样本的分析发现了 24 个不同的白细胞群,与未感染组相比,康复组中 B 细胞、粒细胞和 T 细胞的比例较高。虽然区分粒细胞和巨噬细胞亚群具有挑战性,但对巨噬细胞群中不同基因表达的分析表明,上调的基因与炎症过程有关。特定的T细胞集群显示了cd4-1、cd8a、穿孔素-1和il-2rβ的显著表达,表明存在CD4+T辅助细胞(Th)、CD8+细胞毒性T细胞(Tc)、未成熟T细胞和幼稚T细胞。将 CD4+ T 细胞分为 Th 亚型的尝试缺乏明确的区别,而 CD8+ T 细胞则表现出三个群集,主要是 Tc1 细胞。此外,对康复组和未感染组进行比较后发现,康复组中活化和分泌抗体的 B 细胞比例增加。这项单细胞分析为了解亚洲鲈鱼康复组和未感染组的免疫细胞动态提供了重要信息,为了解亚洲鲈鱼感染 SDDV 后的潜在免疫反应提供了宝贵资料。
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