ODN 2216
目录号 : GC66339
ODN 2216 是一种 TLR9 (toll 样受体 9) 配体或激动剂。ODN 2216 诱导大量的 IFN-α 和 IFN-β。ODN 2216 通过 pDC (浆细胞样 DC) 诱导 IFN-α，并通过 DC (树突状细胞)产生 IL-12 (p40)。ODN 2216 刺激外周血单核细胞 (PBMC) 产生 IFN-γ，这是间接的和由 IFN-α/β 介导的。ODN 2216 可以激活 NK 细胞，促进 IFN-γ 产生 TCR 触发的 CD4+ T 细胞。
Cas No.:332437-00-0
Sample solution is provided at 25 µL, 10mM.
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Quality Control & SDS
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- Purity: >98.00%
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ODN?2216 is a TLR9 (toll-like receptor 9) ligand or agonist. ODN?2216 induces high amounts of IFN-α and IFN-β. ODN 2216 induces IFN-α by pDC (plasmacytoid DC) and IL-12 (p40) production by DC (dendritic cells). ODN 2216 stimulates IFN-γ production in peripheral blood mononuclear cells (PBMC), which is indirect and mediated by IFN-α/β. ODN 2216 can activate NK cells and promote IFN-γ production of TCR-triggered CD4+ T cells[1][2][3][4].
ODN 2216 (6 μg/mL, 9 days) drives Th1 development of naive CD4 T cells[1].
ODN?2216 (2 μM, 15 h) significantly decreased apoptosis of thymic DC[2].
Apoptosis Analysis[2]
| Cell Line: | Thymic DC, pDC |
| Concentration: | 2 μM |
| Incubation Time: | 15 h |
| Result: | Significantly decreased apoptosis of thymic DC (23.1 ± 0.9%), and naturally induced apoptosis of thymic pDC was not decreased (72.8 ± 2.9%). |
ODN 2216 induces T-cell mediated antitumor immune response in murine vaccination models[3]
| Cas No. | 332437-00-0 | SDF | Download SDF |
| 分子式 | 分子量 | 6432 | |
| 溶解度 | 储存条件 | Store at -20°C, away from moisture | |
| General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
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1 mg | 5 mg | 10 mg |
| 1 mM | 0.1555 mL | 0.7774 mL | 1.5547 mL |
| 5 mM | 0.0311 mL | 0.1555 mL | 0.3109 mL |
| 10 mM | 0.0155 mL | 0.0777 mL | 0.1555 mL |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
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| % DMSO % % Tween 80 % saline | ||||||||||
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工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
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[Activation ability of CpG ODNs 2216 on PBMNCs from leukemia patients in remission and killing effect of activated PBMNCs on K562 cells]
Zhongguo Shi Yan Xue Ye Xue Za Zhi 2009 Aug;17(4):874-8.PMID:19698220doi
The aim of this study was to investigate the activation ability of CpG oligodeoxynucleotide (CpG ODN) 2216 on the peripheral blood mononuclear cells (PBMNCs) from leukemia patients in remission and the killing effect of activated PBMNCs on K562 cells. PBMNCs obtained from leukemia patients in remission were incubated with CpG ODN 2216. In control group, PBMNCs were incubated with normal saline (NS). The concentrations of cytokines (IFN-gamma, interleukin-12, interleukin-4, interleukin-10) in culture supernatant of PBMNCs from leukemia patients in remission were analyzed by using ELISA kits. The percentages of Th1, Tc1, Th2, Tc2 cells and killed K562 cells were detected by flow cytometry. The results showed that as compared with control group, CpG ODN 2216 induced higher concentrations production of IFN-gamma, IL-12 in supernatant (p < 0.01). There were no differences in IL-4, IL-10 in supernatant as compared with control group (p > 0.05). The percentages of Th1 and Tc1 cells increased significantly after culture with CpG ODN 2216 as compared with control group (p < 0.05). There was no difference between the percentages of Th2 and Tc2 cells in stimulated group and control group. The killing effect of PBMNCs on K562 cells was significantly different between the stimulated group and control group (p < 0.05). It is concluded that CpG ODNs 2216 can induce strong Th1-like immune activation, with the secretion of type-I cytokine and activation of strong CD8(+) T-cell responses. PBMNCs activated by CpG ODNs can more strongly kill k562 cells in vitro.
Interactions between the Nociceptin and Toll-like Receptor Systems
Cells 2022 Mar 23;11(7):1085.PMID:35406649DOI:10.3390/cells11071085.
Nociceptin and the nociceptin receptor (NOP) have been described as targets for treatment of pain and inflammation, whereas toll-like receptors (TLRs) play key roles in inflammation and impact opioid receptors and endogenous opioids expression. In this study, interactions between the nociceptin and TLR systems were investigated. Human THP-1 cells were cultured with or without phorbol myristate acetate (PMA 5 ng/mL), agonists specific for TLR2 (lipoteichoic acid, LTA 10 µg/mL), TLR4 (lipopolysaccharide, LPS 100 ng/mL), TLR7 (imiquimod, IMQ 10 µg/mL), TLR9 (oligonucleotide (ODN) 2216 1 µM), PMA+TLR agonists, or nociceptin (0.01−100 nM). Prepronociceptin (ppNOC), NOP, and TLR mRNAs were quantified by RT-qPCR. Proteins were measured using flow cytometry. PMA upregulated ppNOC mRNA, intracellular nociceptin, and cell membrane NOP proteins (all p < 0.05). LTA and LPS prevented PMA’s upregulating effects on ppNOC mRNA and nociceptin protein (both p < 0.05). IMQ and ODN 2216 attenuated PMA’s effects on ppNOC mRNA. PMA, LPS, IMQ, and ODN 2216 increased NOP protein levels (all p < 0.05). PMA+TLR agonists had no effects on NOP compared to PMA controls. Nociceptin dose-dependently suppressed TLR2, TLR4, TLR7, and TLR9 proteins (all p < 0.01). Antagonistic effects observed between the nociceptin and TLR systems suggest that the nociceptin system plays an anti-inflammatory role in monocytes under inflammatory conditions.
Effects of CPG ODN on biological behavior of PANC-1 and expression of TLR9 in pancreatic cancer
World J Gastroenterol 2011 Feb 28;17(8):996-1003.PMID:21448350DOI:10.3748/wjg.v17.i8.996.
Aim: To determine the expression of toll-like receptor 9 (TLR9) in pancreatic tumor and the effects of cytosine phosphate-guanosine oligodeoxynucleotides 2216 (CPG ODN2216) on biological behavior of pancreatic carcinoma cell line PANC-1 and explore their clinical significance. Methods: The immunohistochemistry and Western blot were used to determine the expression of TLR9 protein in pancreatic cancer tissues, and immunofluorescence staining was performed to detect the TLR9 protein expression in pancreatic carcinoma cell line PANC-1. To assess the effects of CPG ODN2216 on the invasive property of Panc-1 cells, in vitro cell adhesion, wound-healing scrape, and invasion and cell colony formation were evaluated. Results: TLR9 was highly expressed in pancreatic cancer tissues and PANC-1 cells. The percentage of positive cells expressing TLR9 protein in human pancreatic tissues, paracancerous tissues and normal tissues were 73.3%, 33.3% and 20.0%, respectively, and the protein expression level of TLR9 was gradually descending (P < 0.05). In vitro tests in wound-healing scrape, cell adhesion, colony formation and matrigel invasion showed that the adhesion and motility of PANC-1 cells in CPG ODN 2216 treatment group were significantly lower than in the control group (P < 0.05). The cell growth assay showed that the proliferative ability of PANC-1 cells in treatment group was significantly decreased and CPG ODN2216 had an inhibitive effect in the growth of Panc-1 cells in a dose and time-dependent manner (P < 0.05). Conclusion: The gene of TLR9 is correlated with the invasive and metastatic potential of human pancreatic carcinoma, and CPG ODN2216 induces the inhibition of migration and invasion of Panc-1 cells.
Direct Engagement of TLR9 Ligand with T Helper Cells Leads to Cell Proliferation & Up-regulation of Cytokines
Immunol Invest 2019 Jan;48(1):79-95.PMID:30239236DOI:10.1080/08820139.2018.1515223.
Purpose: Toll like receptor (TLR) engagement is primarily a function of the innate immune cells. The purpose of the study was to assess direct uptake of ODN 2216 in T helper cells and effects on cell proliferation and cytokine expression. Methods: We isolated CD4+ CD25- T helper cells by magnetic sorting and studied the uptake of ODN 2216 using flow cytometry and confocal microscopy. We then studied the effect of ODN 2216 engagement on cell proliferation and cytokine expression using flow cytometry and gene expression of TLR9 signaling genes using real time RT-PCR. Results: We made a chance observation that purified T helper cells from healthy individuals consistently bind to the TLR9 ligand ODN 2216. In PBMCs, on the other hand, 98% of monocytes preferentially bound to ODN 2216 FITC, indicating that they competed with the lymphocytes. We confirmed intracellular localization of ODN 2216 FITC as well as intracellular expression of TLR9 in Thelper cells. Furthermore, ODN 2216 FITC was also co-localized with the lysosomal membrane associated protein 1. The uptake of TLR9 ligand culminated in cellular proliferation, up-regulation of cytokines and increased mRNA expression of TLR9 and IRF7 in T helper cells, in the absence of antigen presenting cells. ODN 2216 uptake was inhibited by promethazine as well as by TLR9 antagonist. Conclusions: Our results show a direct engagement of TLR9 ligand in T helper cells and suggest involvement of TLR9 signalling in CD4+T cells, which may envisage novel targets for TLR inhibitors.
Cytokine induction by immunostimulatory DNA in porcine PBMC is impaired by a hairpin forming sequence motif from the genome of Porcine Circovirus type 2 (PCV2)
Vet Immunol Immunopathol 2011 Feb 15;139(2-4):156-66.PMID:20980058DOI:10.1016/j.vetimm.2010.09.010.
Porcine Circovirus type 2 (PCV2) can cause postweaning multisystemic wasting syndrome (PMWS) in young pigs with severe immunosuppression as a major characteristic of the disease complex. Despite the dramatic involvement of the immune system, the interaction between PCV2 and the host is until date not well understood. The DNA genome of PCV2 contains sequences that in synthetic form (oligodeoxyribonucleotides; ODNs) can act immunomodulatory on porcine peripheral blood mononuclear cells (poPBMCs) in vitro. One such sequence (ODN PCV2/1) acts inhibitory on interferon (IFN)-α production induced by immunostimulatory DNA but not that induced by RNA, and the inhibitory activity is dependent on secondary structure formation. In the present study, the characteristic of ODN PCV2/1 was examined further by altering the nucleotide sequence to disrupt hairpin structure formation but still enable multimer structures through G-tetrads. This modification resulted in loss of IFN-α-inhibitory activity of the ODN and thus indicated the importance of hairpin structures. In addition, ODN PCV2/1 was compared to another inhibitory ODN (IRS 869) previously used in human and murine cells. In contrast to ODN PCV2/1, ODN IRS 869 did not inhibit IFN-α production induced by class A ODN 2216 but was a more efficient inhibitor of IFN-α production induced by plasmid DNA than ODN PCV2/1. In cultures induced by the RNA stimulator Poly I:C, however, a strong synergistic IFN-α stimulatory effect was seen in combination with ODN IRS 869. These results indicate that ODN PCV2/1 and ODN IRS 869 function through separate mechanisms to affect cytokine production by immune cells. The effect of ODN PCV2/1 was studied further by monitoring the expression of mRNA for IFN-α, IL-12p40, IL-10, IL-6, IFN-γ, IL-1β, TGF-β, and TNF-α in cultures of poPBMC stimulated with ODN 2216 or Poly I:C. Results from qPCR analyses showed that ODN PCV2/1 clearly inhibited the expression of IFN-α, IL-12p40, IL-10 and IL-6 when induced by ODN 2216, but did not seem to affect any of the cytokines examined when induced by Poly I:C. Initial studies using confocal microscopy and fluorochrome labelled ODNs indicate that ODN 2216 and ODN PCV2/1 co-localize in subpopulations of poPBMC.