温度对牙鲆（）mIgM B 淋巴细胞抗病毒反应的影响。
The Influence of Temperature on the Antiviral Response of mIgM B Lymphocytes Against in Flounder ().
机构信息
Laboratory of Pathology and Immunology of Aquatic Animals, Key Laboratory of Mariculture, Ministry of Education (KLMME), Ocean University of China, Qingdao, China.
Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao, China.
出版信息
Front Immunol. 2022 Feb 7;13:802638. doi: 10.3389/fimmu.2022.802638. eCollection 2022.
(HIRRV) is an ongoing threat to the aquaculture industry. The water temperature for the onset of HIRRV is below 15°C, the peak is about 10°C, but no mortality is observed over 20°C. Previous studies found the positive signal of matrix protein of HIRRV (HIRRV-M) was detected in the peripheral blood leukocytes of viral-infected flounder. Flow cytometry and indirect immunofluorescence assay showed that HIRRV-M was detected in mIgM B lymphocytes in viral-infected flounder maintained at 10°C and 20°C, and 22% mIgM B lymphocytes are infected at 10°C while 13% are infected at 20°C, indicating that HIRRV could invade into mIgM B lymphocytes. Absolute quantitative RT-PCR showed that the viral copies in mIgM B lymphocytes were significantly increased at 24 h post infection (hpi) both at 10°C and 20°C, but the viral copies in 10°C infection group were significantly higher than that in 20°C infection group at 72 hpi and 96 hpi. Furthermore, the B lymphocytes were sorted from HIRRV-infected flounder maintained at 10°C and 20°C for RNA-seq. The results showed that the differentially expression genes in mIgM B lymphocyte of healthy flounder at 10°C and 20°C were mainly enriched in metabolic pathways. Lipid metabolism and Amino acid metabolism were enhanced at 10°C, while Glucose metabolism was enhanced at 20°C. In contrast, HIRRV infection at 10°C induced the up-regulation of the Complement and coagulation cascades, FcγR-mediated phagocytosis, Platelets activation, Leukocyte transendothelial migration and Natural killer cell mediated cytotoxicity pathways at 72 hpi. HIRRV infection at 20°C induced the up-regulation of the Antigen processing and presentation pathway at 72 hpi. Subsequently, the temporal expression patterns of 16 genes involved in Antigen processing and presentation pathway were investigated by qRT-PCR, and results showed that the pathway was significantly activated by HIRRV infection at 20°C but inhibited at 10°C. In conclusion, HIRRV could invade into mIgM B lymphocytes and elicit differential immune response under 10°C and 20°C, which provide a deep insight into the antiviral response in mIgM B lymphocytes.
（HIRRV）是水产养殖业的持续威胁。HIRRV 的发病水温低于 15°C，高峰期约为 10°C，但在 20°C 以上不会观察到死亡率。先前的研究发现，HIRRV 的基质蛋白（HIRRV-M）的阳性信号在受病毒感染的比目鱼外周血白细胞中被检测到。流式细胞术和间接免疫荧光检测显示，在 10°C 和 20°C 下维持的受病毒感染的比目鱼中，HIRRV-M 被检测到在 mIgM B 淋巴细胞中，10°C 时 22%的 mIgM B 淋巴细胞被感染，而 20°C 时 13%的 mIgM B 淋巴细胞被感染，表明 HIRRV 能够侵入 mIgM B 淋巴细胞。绝对定量 RT-PCR 显示，在感染后 24 小时（hpi），10°C 和 20°C 时 mIgM B 淋巴细胞中的病毒拷贝数均显著增加，但在 72 hpi 和 96 hpi 时，10°C 感染组的病毒拷贝数明显高于 20°C 感染组。此外，从维持在 10°C 和 20°C 的 HIRRV 感染比目鱼中分离出 B 淋巴细胞进行 RNA-seq。结果表明，10°C 和 20°C 时健康比目鱼 mIgM B 淋巴细胞中的差异表达基因主要富集在代谢途径中。10°C 时，脂质代谢和氨基酸代谢增强，而 20°C 时葡萄糖代谢增强。相比之下，10°C 时 HIRRV 感染诱导补体和凝血级联、FcγR 介导的吞噬作用、血小板激活、白细胞穿过内皮迁移和自然杀伤细胞介导的细胞毒性途径在 72 hpi 时上调。20°C 时 HIRRV 感染诱导抗原加工和呈递途径在 72 hpi 时上调。随后，通过 qRT-PCR 研究了参与抗原加工和呈递途径的 16 个基因的时间表达模式，结果表明，20°C 时 HIRRV 感染显著激活该途径，但在 10°C 时抑制该途径。总之，HIRRV 可以在 10°C 和 20°C 下侵入 mIgM B 淋巴细胞，并引发不同的免疫反应，这为 mIgM B 淋巴细胞中的抗病毒反应提供了深入的了解。
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