CpG ODN 2102 促进抗菌免疫反应，并增强金黄鰤（Trachinotus ovatus）疫苗诱导的保护作用。

CpG ODN 2102 promotes antibacterial immune responses and enhances vaccine-induced protection in golden pompano (Trachinotusovatus).

机构信息

Sanya Nanfan Research Institute, Hainan University, Sanya, 572022, China; State Key Laboratory of Marine Resource Utilization in South China Sea, Hainan University, Haikou, 570228, China; Collaborative Innovation Center of Marine Science and Technology, Hainan University, Haikou, 570228, China.

Sanya Nanfan Research Institute, Hainan University, Sanya, 572022, China; Collaborative Innovation Center of Marine Science and Technology, Hainan University, Haikou, 570228, China.

出版信息

Fish Shellfish Immunol. 2023 Jun;137:108783. doi: 10.1016/j.fsi.2023.108783. Epub 2023 May 1.

DOI:10.1016/j.fsi.2023.108783

PMID:37137380

Abstract

CpG oligodeoxynucleotides (ODNs) are oligodeoxynucleotides containing CpG motifs and can be recognized by toll-like receptor 9 (TLR9), activating the host's immune responses. In this study, ten different CpG ODNs were designed and synthesized to study the antibacterial immune responses of CpG ODNs in golden pompano (Trachinotus ovatus). Results showed that CpG ODN 2102 significantly improved the immunity of golden pompano against bacteria. Besides, CpG ODN 2102 promoted the proliferation of head kidney lymphocytes and activated the head kidney macrophages. When TLR9-specific small interfering RNA (siRNA) was used to interfere with TLR9 expression, the immune responses were decreased. Moreover, the expression levels of myeloid differentiation primary response 88 (Myd88), p65, tumor necrosis factor receptor-associated factor 6 (TRAF6), and tumor necrosis factor-alpha (TNF-α) in the TLR9-knockdown golden pompano kidney (GPK) cells were significantly reduced. The nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) promoter activity of the TLR9-knockdown GPK cells was also significantly reduced. In vivo, the antibacterial immune effects induced by CpG ODN 2102 in golden pompano were mostly abolished when TLR9 expression was knocked down. These results suggested that TLR9 was involved in the immune responses induced by CpG ODN 2102. CpG ODN 2102 also enhanced the protective effect of the Vibrio harveyi vaccine pCTssJ, where the survival rate of golden pompano was significantly improved by 20%. In addition, CpG ODN 2102 enhanced the messenger RNA (mRNA) expression levels of TLR9, Myxovirus resistance (Mx), interferon γ (IFN-γ), TNF-α, interleukin (IL)-1β, IL-8, major histocompatibility complex class (MHC) Iα, MHC IIα, Immunoglobulin D (IgD), and IgM. Therefore, TLR9 was involved in the antibacterial immune responses induced by CpG ODN 2102 and CpG ODN 2102 possessed adjuvant immune effects. These results enlarged our knowledge of the antibacterial immunity of fish TLRs signaling pathway and had important implications for exploring natural antibacterial molecules in fish and developing new vaccine adjuvants.

摘要

CpG 寡脱氧核苷酸（ODN）是含有 CpG 基序的寡脱氧核苷酸，可以被 Toll 样受体 9（TLR9）识别，激活宿主的免疫反应。在这项研究中，设计并合成了十种不同的 CpG ODN，以研究 CpG ODN 在金黄鯵（Trachinotus ovatus）中的抗菌免疫反应。结果表明，CpG ODN 2102 显著提高了金黄鯵对细菌的免疫力。此外，CpG ODN 2102 促进了头肾淋巴细胞的增殖，并激活了头肾巨噬细胞。当使用 TLR9 特异性小干扰 RNA（siRNA）干扰 TLR9 表达时，免疫反应会降低。此外，TLR9 敲低金黄鯵肾脏（GPK）细胞中髓样分化初级反应 88（Myd88）、p65、肿瘤坏死因子受体相关因子 6（TRAF6）和肿瘤坏死因子-α（TNF-α）的表达水平显著降低。TLR9 敲低 GPK 细胞的核因子 kappa-轻链增强子的 B 细胞激活因子（NF-κB）启动子活性也显著降低。在体内，当 TLR9 表达被敲低时，CpG ODN 2102 在金黄鯵中诱导的抗菌免疫效应大部分被消除。这些结果表明 TLR9 参与了 CpG ODN 2102 诱导的免疫反应。CpG ODN 2102 还增强了哈维氏弧菌疫苗 pCTssJ 的保护作用，使金黄鯵的存活率提高了 20%。此外，CpG ODN 2102 增强了 TLR9、流感病毒抗性（Mx）、干扰素 γ（IFN-γ）、TNF-α、白细胞介素（IL）-1β、IL-8、主要组织相容性复合体 Iα、MHC IIα、免疫球蛋白 D（IgD）和 IgM 的信使 RNA（mRNA）表达水平。因此，TLR9 参与了 CpG ODN 2102 诱导的抗菌免疫反应，CpG ODN 2102 具有佐剂免疫效应。这些结果扩大了我们对鱼类 TLRs 信号通路抗菌免疫的认识，对探索鱼类天然抗菌分子和开发新型疫苗佐剂具有重要意义。