Flow cytometic analysis of Penaeus monodon haemocyte responses to poly I:C
Flow cytometic analysis of Penaeus monodon haemocyte responses to poly I:C
复制标题
流式细胞术分析斑节对虾血细胞对 Poly I:C 的反应
DOI：
10.1016/j.fsi.2017.12.045
复制
发表时间：
2018-03-01
影响因子：
4.7
通讯作者：
Zheng, Pei-Hua
中科院分区：
文献类型：
--
作者：
Xian, Jian-An;Zhang, Xiu-Xia;Zheng, Pei-Hua
本研究旨在利用流式细胞术研究单对虾血细胞对poly I:C刺激的细胞反应。测定了聚I: c注射对虾的总血细胞计数（THC）、不同血细胞亚群[透明细胞（HC）、半颗粒细胞（SGC）和颗粒细胞（GC）]百分比、非特异性酯酶活性（EA）、总活性氧/活性氮（ROS/RNS）产量、一氧化氮（NO）产量、凋亡血细胞比率和血浆酚氧化酶（PO）活性。结果表明，低剂量（5 μ g对虾（-1））poly I:C可显著提高THC、GC比例、ROS/RNS产量和NO产量，但对EA、细胞凋亡和PO活性无显著影响。在高剂量（20 μ g对虾（-1））注射poly I:C的早期，也可以观察到THC和GC比例的提高，提示GC可能被诱导从造血或其他组织中释放出来参与免疫反应，该亚群可能是参与细胞防御病毒的主要细胞类型。在后期，GC和SGC的比例随着THC的减少而平行减少，表明GC和SGC的消耗主要是导致循环血细胞计数减少的原因。高剂量poly I:C刺激对虾血细胞中ROS/RNS的产生和NO的产生均明显增加，而EA的升高和PO活性的抑制均不明显，提示ROS/RNS依赖系统在对虾的病毒免疫防御中起重要作用。另一方面，在ROS/RNS和NO产生急剧增加后，凋亡血细胞比例增加，THC减少，提示受刺激的ROS/RNS可能过量且有害，是导致血细胞凋亡和耗竭的主要因素。注射48 h后THC恢复，而血细胞凋亡也恢复到控制水平，提示细胞凋亡可能有助于清除受损、虚弱或感染的血细胞，更新循环中的血细胞，可以认为它是一种重要的防御病毒的策略。
This study was aimed at investigating the cellular responses of Penaeus monodon haemocytes to poly I:C stimulation using flow cytometric assay. Total haemocyte count (THC), percentages of different haemocyte sub populations [hyaline cells (HC), semigranular cells (SGC) and granular cells (GC)], non-specific esterase activity (EA), total reactive oxygen species/reactive nitrogen species (ROS/RNS) production, nitric oxide (NO) production, apoptotic haemocyte ratio and plasmic phenoloxidase (PO) activity were determined in poly I:C-injected shrimp. Results showed that poly I:C at a low dose (5 mu g shrimp(-1)) caused obvious increases in THC, GC proportion, ROS/RNS production and NO production, but had no significant effect on EA, apoptosis and PO activity. In the early stage of poly I:C injection at a high dose (20 mu g shrimp(-1)), THC and GC proportion improvements could also be observed, suggesting that GC might be induced to release from hemocytopoietic or other tissues to participate in immune response, and this subpopulation might be the main cell type involved in the cellular defence against virus. In the later period, proportions of both GC and SGC reduced paralleled by THC reduction, indicating that depletion of GC and SGC was mainly contributed to the reduced count of circulating haemocyte. Obvious increases in ROS/RNS production and NO production were induced in haemocyte of shrimp under a high dose of poly I:C stimulation, but only slight rise of EA and suppression of PO activity could be observed in poly I:C-stimulated shrimp, suggesting that ROS/RNS-dependent system was vital in the immune defence of shrimp against virus. On the other hand, increase of apoptotic haemocyte ratio and THC reduction were presented after the drastic increases of ROS/RNS and NO productions, implying that the stimulated ROS/RNS might be excess and harmful, and was the major factor for the haemocyte apoptosis and depletion. THC recovered after 48 h injection, while haemocyte apoptosis also returned to the control level, suggesting that apoptosis might be contributed to eliminate damaged, weak or infected haemocytes to renew the circulating haemocytes, and it could be considered as an important defending strategy against virus.