College of Fisheries and Life Science, Dalian Ocean University, Dalian, 11026, China.
College of Fisheries and Life Science, Dalian Ocean University, Dalian, 11026, China.
Fish Shellfish Immunol. 2022 Dec;131:1206-1213. doi: 10.1016/j.fsi.2022.11.023. Epub 2022 Nov 17.
CpG oligodeoxynucleotides (ODN), as an effective adjuvant or immunopotentiator, activate the immune system and induce various immune responses. Recently, it has also been reported that high dose of CpG ODN can lead to immunosuppression. However, the underlying mechanism of CpG ODN-mediated immune response remains largely unknown in invertebrates. In the present study, the role of ERK in regulating expression levels of anti-lipopolysaccharide factors (ALFs) induced by different doses of CpG ODN 2395 was analyzed in Chinese mitten crab, Eriocheir sinensis. The mRNA expression levels of EsALFs (EsALF1, EsALF2 and EsALF3) and EsERK in haemocytes were observed to increase from 6 h to 48 h post low doses of CpG ODN 2395 (0.5 μg and 2.5 μg) stimulation, while they were suppressed after high dose of CpG ODN 2395 (12.5 μg) injection. Meanwhile, the phosphorylation levels of ERK in haemocytes were significantly promoted after low doses of CpG ODN 2395 injection, and a reduce level of ERK phosphorylation was observed after high dose of CpG ODN 2395 injection. Further investigation showed that the expression levels of EsALFs induced by CpG ODN 2395 were markedly down-regulated after knocking down the expression of EsERK. Similarly, the EsALFs mRNA expression were also inhibited post different doses of CpG ODN 2395 stimulation in PD98059 (ERK inhibitor) injection crabs. These results collectively suggest that ERK is involved in regulating the expression level of EsALFs induced by different dose of CpG ODN 2395 in Chinese mitten crab, which contribute to the understanding of the regulation of CpG ODN involving in immune response in crustaceans.
CpG 寡脱氧核苷酸(ODN)作为一种有效的佐剂或免疫增强剂,可激活免疫系统并诱导各种免疫反应。最近,也有报道称高剂量的 CpG ODN 可导致免疫抑制。然而,在无脊椎动物中,CpG ODN 介导的免疫反应的潜在机制在很大程度上仍不清楚。本研究分析了 CpG ODN 2395 不同剂量在中华绒螯蟹(Eriocheir sinensis)中诱导抗脂多糖因子(ALFs)表达水平时 ERK 的作用。观察到低剂量 CpG ODN 2395(0.5μg 和 2.5μg)刺激后 6 h 至 48 h 血细胞中 EsALFs(EsALF1、EsALF2 和 EsALF3)和 EsERK 的 mRNA 表达水平增加,而高剂量 CpG ODN 2395(12.5μg)注射后则受到抑制。同时,低剂量 CpG ODN 2395 注射后,血细胞中 ERK 的磷酸化水平显著升高,而高剂量 CpG ODN 2395 注射后则观察到 ERK 磷酸化水平降低。进一步研究表明,敲低 EsERK 的表达后,CpG ODN 2395 诱导的 EsALFs 表达水平明显下调。同样,在 PD98059(ERK 抑制剂)注射螃蟹中,不同剂量的 CpG ODN 2395 刺激后 EsALFs mRNA 表达也受到抑制。这些结果共同表明,ERK 参与调节 CpG ODN 2395 不同剂量诱导的 EsALFs 表达水平,有助于理解 CpG ODN 参与甲壳动物免疫反应的调节。