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| 黄条鰤白介素-1β基因克隆及其免疫应答分析 |
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王琳,王滨,徐永江,关长涛
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1.农业农村部海洋渔业可持续发展重点实验室(中国水产科学研究院黄海水产研究所) 山东 青岛 266071;2.崂山实验室海洋渔业科学与食物产出过程功能研究室 山东 青岛 266237;3.崂山实验室海洋渔业科学与食物产出过程功能研究室 山东 青岛 266238;4.崂山实验室海洋渔业科学与食物产出过程功能研究室 山东 青岛 266239;5.崂山实验室海洋渔业科学与食物产出过程功能研究室 山东 青岛 266240
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| 摘要: |
| 白介素-1β是一种典型的促炎细胞因子，参与调控免疫细胞增殖、分化和凋亡等过程。本研究从黄条鰤(Seriola aureovittata)中鉴定到2个白介素-1β分子(分别命名为SaIL-1β1和SaIL-1β2)。SaIL-1β1全长cDNA序列为1 292 bp，开放阅读框长度为828 bp，编码275个氨基酸；SaIL-1β2 cDNA序列为1 337 bp，开放阅读框长度为960 bp，编码319个氨基酸。SaIL-1β1和SaIL-1β2编码的蛋白均含有IL-1保守的结构域和12个β折叠，具有结构上的保守性。组织表达分布显示，SaIL-1β1在头肾中表达量最高，脾脏和肝脏次之；而SaIL-1β2在鳃中表达量最高，头肾和脾脏次之。脂多糖(LPS)刺激后，SaIL-1β1和SaIL-1β2在头肾和脾脏中的表达量均显著增加。在头肾中，LPS刺激后6 h，SaIL-1β1急剧上升至对照组的10.03倍(P<0.05)，随后逐渐回落，在12、24、48、72 h分别为对照组的7.15、4.09、2.71、3.03倍(P<0.05)；在刺激后6 h，SaIL-1β2表达量急剧上升至对照组的11.49 倍(P<0.05)，最后逐渐回落，48 h恢复至正常水平，72 h下降至对照组的0.29倍(P<0.05)。脾脏中，LPS刺激后6 h，SaIL-1β1表达量急剧上升至对照组的6.59倍(P<0.05)，随后逐渐回落；SaIL-1β2转录水平表达模式与SaIL-1β2相似。综上，本研究在黄条鰤中鉴定了2种白介素-1β分子，并探讨了其在免疫应答中的表达规律，为研究白介素-1β分子在黄条鰤抗菌免疫中的作用提供了基础。 |
| 关键词: 白介素-1β 免疫应答 细胞因子 黄条鰤 |
| DOI：10.19663/j.issn2095-9869.20220418002 |
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| Two putative interleukin-1 beta molecules involved in the immune response of Seriola aureovittata |
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WANG Lin1,2, WANG Bin3,4, XU Yongjiang5,6, GUAN Changtao7,8
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1.Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture and Rural Affairs, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China;2.Laboratory for Marine Fisheries Science and Food Production Processes, Laoshan Laboratory, Qingdao 266237, China;3.Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture and Rural Affairs, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266072, China;4.Laboratory for Marine Fisheries Science and Fo2 晦㽦 晦㽦 蘃覽ऀ耀ﳞ䑨Ḫ䑥�대 㾀ዮ Āʨ 㾀㨀耀 ;5.Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture and Rural Affairs, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266073, China;6.Laboratory for Marine Fisheries Science and Fo ᎓妚䁮 㿠 㿠 㿠 㿠 㾀 㾀 㾀 㾀 㾀 ;7.Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture and Rural Affairs, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266074, China;8.Laboratory for Marine Fisheries Science and Foﷺ﷽ﺂﺂﺄﺄﺆﺆﺈﺈﺊﺌﺎﺎﺐﺒﺔﺔﺖﺘﺚﺜﺞﺠﺢﺤﺦﺨﺪﺪﺬﺬﺮﺮﺰﺰﺲﺴﺶﺸﺺﺼﺾﻀﻂﻄﻆﻈﻊﻌﻎﻐﻒﻔﻖﻘﻚﻜﻞﻠﻢﻤﻦﻬﻮﻮﻰﻰ
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| Abstract: |
| Interleukin-1 beta (IL-1β) is the quintessential pro-inflammatory cytokine, playing important roles in immune cell proliferation, differentiation, and apoptosis. The IL-1β genes have been characterized for many fish species. Unlike mammalian genes, several species of fish possess two IL-1β genes, which may be a consequence of genome duplication in particular fish lineages. Yellowtail kingfish (Seriola aureovittata) is a pelagic marine finfish species, which is an emerging candidate for the aquaculture industry. Therefore, details encompassing the role of IL-1β in the immune response aids a development strategy for economic and efficient aquaculture. In the present study, two novel il-1β molecules were identified from S. aureovittata (designated as SaIL-1β1 and SaIL-1β2). The full-length cDNA of SaIL-1β1 was 1 292 bp with a 828 bp open reading frame, encoding a polypeptide of 275 amino acids, while the full-length cDNA of SaIL-1β2 was 1 337 bp with a 960 bp open reading frame, encoding a polypeptide of 319 amino acids. Both SaIL-1β molecules contain an IL1 domain, 12 β-sheets, and a C-terminal conserved region, which are IL-1 family signature characters. A phylogenetic analysis revealed the fish IL-1βs clustered together. SaIL-1β1 and IL-1β in Seriola dumerili initially clustered together. However, SaIL-1β2 initially clustered with IL-1β in Trachinotus ovatu. Real-time PCR showed the transcripts of SaIL-1β1 and SaIL-1β2 were present in all the tested tissues, including the head kidney, spleen, liver, gill, heart, stomach, pituitary gland, muscle, and brain. Among them, the SaIL-1β1 transcripts were predominantly in the head kidney, spleen, and liver. The expression of SaIL-1β2 mRNA was predominantly in the gill, head kidney, and spleen. The high expression of SaIL-1β1 and SaIL-1β2 mRNA in the immune related organs implies a potential role in immune regulation. LPS is a pro-inflammatory endotoxin used as a standard immune activating agent. After LPS stimulation, the two SaIL-1βs transcripts were vigorously altered in the head kidney and spleen. SaIL-1β1 transcripts were significantly increased at 6 h, 12 h, 24 h, 48 h, and 72 h post-stimulation in the head kidney (10.03, 7.15, 4.09, 2.71, and 3.03-fold of the control group results, respectively) (P<0.05). Meanwhile, SaIL-1β2 transcripts significantly increased from 6 h to 24 h post-stimulation after infection in the head kidney (11.49, 4.08, and 4.70-fold of the control group, respectively) (P<0.05), had returned to normal at 48 h, and had decreased at 72 h (to 0.29-fold of the control group) (P<0.05). In the spleen, SaIL-1β1 transcripts were sharply elevated at 6 h (to 6.59-fold of control group), gradually returned to normal at 12 h, 24 h, and 48 h (3.85, 4.09, 2.17, and 2.65-fold of control group, respectively) (P<0.05), and had dropped to basal level by 72 h. SaIL-1β2 mRNA had a similar expression pattern to SaIL-1β1. SaIL-1β2 mRNA increased from 6 h to 48 h post-stimulation after infection (7.25, 3.20, 1.59, and 1.59-fold of the control group, respectively) (P<0.05) and had returned to normal by 72 h. The activated immune signaling promoted the expression of SaIL-1β1and SaIL-1β2 in the immune response, especially in the early stage, indicating they might be a pro-inflammatory cytokine in S. aureovittata. Collectively, the conserved structure and tissue distribution of SaIL-1β1 and SaIL-1β2, together with their sensitivity to LPS stimulation suggests their involvement in the immune response, providing clues to our understanding of the role of IL-1β in S. aureovittata during immune response. |
| Key words: Interleukin-1 beta Immune response Cytokine Seriola aureovittata |
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