Taju G, Madan N, Abdul Majeed S, Kumar T Raj, Thamizhvanan S, Otta S K, Sahul Hameed A S
OIE Reference Laboratory for WTD, PG & Research Department of Zoology, C. Abdul Hakeem College, Melvisharam, Vellore District, Tamilnadu, India.
J Fish Dis. 2015 May;38(5):451-65. doi: 10.1111/jfd.12256. Epub 2014 Jun 10.
In this study, dsRNA specific to VP28 gene of white spot syndrome virus (WSSV) of shrimp was synthesized in Escherichia coli in large scale and studied the immune response of shrimp to dsRNA-VP28. The haematological parameters such as clotting time and total haemocytes counts, and immunological parameters such as prophenoloxidase (proPO), superoxide dismutase (SOD), superoxide anion (SOA) and malondialdehyde content, as well as the mRNA expression of ten immune-related genes were examined to estimate the effect of dsRNA-VP28 on the innate immunity of Litopenaeus vannamei. The activities of proPO, SOA and SOD significantly increased in haemocyte after dsRNA-VP28 treatment, whereas MDA content did not change significantly. Among the ten immune-related genes examined, only the mRNA expression of proPO, cMnSOD, haemocyanin, crustin, BGBP, lipopolysaccharides (LPs), lectin and lysozyme in haemocytes, gill and hepatopancreas of L. vannamei, was significantly upregulated at 12 h after dsRNA-VP28 treatment, while no significant expression changes were observed in Toll receptor and tumour receptor genes. The increase of proPO and SOD activities, and SOA level and mRNA expression level of proPO, cMnSOD, haemocyanin, crustin, BGBP, LPs, lectin and lysozyme after dsRNA-VP28 stimulation indicate that these immune-related genes were involved in dsRNA-VP28-induced innate immunity in shrimp.
在本研究中,在大肠杆菌中大规模合成了对虾白斑综合征病毒(WSSV)VP28基因特异的双链RNA(dsRNA),并研究了虾对dsRNA-VP28的免疫反应。检测了血液学参数如凝血时间和总血细胞计数,以及免疫学参数如前酚氧化酶(proPO)、超氧化物歧化酶(SOD)、超氧阴离子(SOA)和丙二醛含量,以及十个免疫相关基因的mRNA表达,以评估dsRNA-VP28对凡纳滨对虾先天免疫的影响。dsRNA-VP28处理后,血细胞中proPO、SOA和SOD的活性显著增加,而MDA含量没有显著变化。在所检测的十个免疫相关基因中,dsRNA-VP28处理12小时后,仅凡纳滨对虾血细胞、鳃和肝胰腺中proPO、cMnSOD、血蓝蛋白、甲壳素、BGBP、脂多糖(LPs)、凝集素和溶菌酶的mRNA表达显著上调,而Toll受体和肿瘤受体基因未观察到显著的表达变化。dsRNA-VP28刺激后proPO和SOD活性、SOA水平以及proPO、cMnSOD、血蓝蛋白、甲壳素、BGBP、LPs、凝集素和溶菌酶的mRNA表达水平的增加表明,这些免疫相关基因参与了dsRNA-VP28诱导的虾先天免疫。