Separation and phagocytosis analysis of peripheral blood leukocytes from Qihe crucian carp Carassius auratus

Zhu, Lei; Zhao, Tong; Wang, Xinru; Yang, Shun Fa; Hou, Libo; Li, Chen; Jiang, Xiaolong; Zhang, Jie; Zhao, Xianliang; Pei, Chao; Li, Li; Kong, Xianghui

doi:10.1016/j.aquaculture.2022.737992

Cited by 2 publications

(2 citation statements)

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“…In a previous study, a rapid prominent leukocytosis in the blood of KHV-infected carp occurred during the initial two dpi and throughout the 14 days of a clinical infection at 25 °C [43]. Flow cytometric identification of leukocyte populations was based on previous studies on carp blood leukocytes [20]. Here, leukocytosis was observed as a significant increase in P1 at ten and P2 at , number of fish were three KHV-infected fish for the RNA-Seq analysis and four KHV-infected and control carps for the RT-qPCR tests, all sampled at ten days post infection.…”

Section: Discussionmentioning

confidence: 99%

“…Blood leukocytes were separated using density gradient centrifugation following [ 20 ]. Briefly, equal volumes of blood and phosphate-buffered saline (PBS) (Merck, Gillingham, UK) were mixed and layered into a freshly made discontinuous (from the top layer down) 20%, 30%, 40%, and 50% density gradient of 1.130 g/mL Percoll ® , pH 8.5–9.5 (Merck, UK) at a ratio of 1 mL of diluted blood in 6 mL.…”

Section: Methodsmentioning

confidence: 99%

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Transcriptomic Responses to Koi Herpesvirus in Isolated Blood Leukocytes from Infected Common Carp

Cano,

Blaker,

Hartnell

et al. 2024

Viruses

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Koi herpesvirus (KHV, CyHV-3) causes severe economic losses in carp farms. Its eradication is challenging due to the establishment of latency in blood leukocytes and other tissues. To understand the molecular mechanisms leading to KHV infection in leukocytes, common carp were bath-exposed to KHV at 17 °C. After confirming the presence of viral transcripts in blood leukocytes at ten days post infection, RNA-Seq was performed on peripheral blood leukocytes on the Illumina NovaSeq. KHV infection triggered a robust immune response mediated by pattern recognition receptors, mainly toll-like receptors (tlr2, tlr5, tlr7, and tlr13), urokinase plasminogen activator surface receptor-like, galectin proteins, and lipid mediators such as leukotriene B4 receptor 1. Enriched pathways showed increased mitochondria oxidative phosphorylation and the activation of signalling pathways such as mitogen-activated protein kinases (MAPKs) and vascular endothelial growth factor (VEGF). KHV-infected leukocytes showed low production of reactive oxygen species (ROS) and glutathione metabolism, high iron export and phagocytosis activity, and low autophagy. Macrophage polarization was deduced from the up-regulation of genes such as arginase non-hepatic 1-like, macrophage mannose receptor-1, crem, il-10, and il-13 receptors, while markers for cytotoxic T cells were observed to be down-regulated. Further work is required to characterise these leukocyte subsets and the molecular events leading to KHV latency in blood leukocytes.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%