scholarly journals Toll-Like Receptor 21 of Chicken and Duck Recognize a Broad Array of Immunostimulatory CpG-oligodeoxynucleotide Sequences

Vaccines ◽  
2020 ◽  
Vol 8 (4) ◽  
pp. 639
Author(s):  
Yu-Chen Chuang ◽  
Jen-Chih Tseng ◽  
Jing-Xing Yang ◽  
Yi-Ling Liu ◽  
Da-Wei Yeh ◽  
...  
Keyword(s):  
Nucleotide Sequence ◽  
Mammalian Cells ◽  
Cpg Odn ◽  
Toll Like Receptor ◽  
Cpg Oligodeoxynucleotides ◽  
Sequence Recognition ◽  
Cpg Oligodeoxynucleotide ◽  
Cpg Odns ◽  
Specific Activities ◽  
Species Specific

CpG-oligodeoxynucleotides (CpG-ODNs) mimicking the function of microbial CpG-dideoxynucleotides containing DNA (CpG-DNA) are potent immune stimuli. The immunostimulatory activity and the species-specific activities of a CpG-ODN depend on its nucleotide sequence properties, including CpG-hexamer motif types, spacing between motifs, nucleotide sequence, and length. Toll-like receptor (TLR) 9 is the cellular receptor for CpG-ODNs in mammalian species, while TLR21 is the receptor in avian species. Mammalian cells lack TLR21, and avian cells lack TLR9; however, both TLRs are expressed in fish cells. While nucleotide sequence properties required for a CpG-ODN to strongly activate mammalian TLR9 and its species-specific activities to different mammalian TLR9s are better studied, CpG-ODN activation of TLR21 is not yet well investigated. Here we characterized chicken and duck TLR21s and investigated their activation by CpG-ODNs. Chicken and duck TLR21s contain 972 and 976 amino acid residues, respectively, and differ from TLR9s as they do not have an undefined region in their ectodomain. Cell-based TLR21 activation assays were established to investigate TLR21 activation by different CpG-ODNs. Unlike grouper TLR21, which was preferentially activated by CpG-ODN with a GTCGTT hexamer motif, chicken and duck TLR21s do not distinguish among different CpG-hexamer motifs. Additionally, these two poultry TLR21s were activated by CpG-ODNs with lengths ranging from 15 to 31 nucleotides and with different spacing between CpG-hexamer motifs. These suggested that compared to mammalian TLR9 and grouper TLR21, chicken and duck TLR21s have a broad CpG-ODN sequence recognition profile. Thus, they could also recognize a wide array of DNA-associated molecular patterns from microbes. Moreover, CpG-ODNs are being investigated as antimicrobial agents and as vaccine adjuvants for different species. This study revealed that there are more optimized CpG-ODNs that can be used in poultry farming as anti-infection agents compared to CpG-ODN choices available for other species.

scholarly journals ADP-Ribosylation Factor 3 Mediates Cytidine-Phosphate-Guanosine Oligodeoxynucleotide-Induced Responses by Regulating Toll-Like Receptor 9 Trafficking

2015 ◽  
Vol 7 (6) ◽  
pp. 623-636 ◽  
Author(s):  
Jing-Yiing Wu ◽  
Cheng-Chin Kuo
Keyword(s):  
Protein Trafficking ◽  
Dominant Negative ◽  
Dominant Negative Mutant ◽  
Induced Responses ◽  
Cpg Odn ◽  
Toll Like Receptor ◽  
Adp Ribosylation ◽  
Cpg Oligodeoxynucleotide ◽  
Adp Ribosylation Factor ◽  
Constitutively Active

Toll-like receptor 9 (TLR9) trafficking from the endoplasmic reticulum (ER) into endolysosomes is critical for eliciting cytidine-phosphate-guanosine (CpG) DNA-mediated immune responses. ADP-ribosylation factor 3 (ARF3) is a member of the Ras superfamily, which is crucial for a wide variety of cellular events including protein trafficking. In this study, we found that the inhibition of ARF3 by dominant mutants and siRNA impaired CpG oligodeoxynucleotide (ODN)-mediated responses whereas cells expressing the constitutively active ARF3 mutant enhanced CpG ODN-induced NF-κB activation and cytokine production. Further experiments with MyD88-overexpressing fibroblast cells transfected with a dominant-negative mutant and a constitutively active mutant of ARF3 demonstrated that ARF3 regulated CpG ODN-mediated signaling upstream of MyD88. Additional studies have shown that ARF3 inhibition impairs TLR9 trafficking from the ER into endolysosomes, thereby inhibiting the functional cleavage of TLR9, although it has no significant effect on CpG ODN uptake. Furthermore, activated ARF3 is associated with Unc93B1 and TLR9, suggesting that ARF3 conducts TLR9 trafficking by forming the TLR9-Unc93B1-ARF3 complex. Overall, our findings demonstrate that a novel ARF3 axis pathway mediates CpG ODN-induced responses by regulating TLR9 trafficking.

Stimulation of B-CLL Cells with Interleukin 21 and Toll Like Receptor Agonists Induces a CTL-Like Transcriptional Profile.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 3685-3685
Author(s):  
Magdalena Hagn ◽  
Verena Ebel ◽  
Kai Sontheimer ◽  
Thamara Beyer ◽  
Sue E. Blackwell ◽  
...  
Keyword(s):  
Transcription Factors ◽  
B Cells ◽  
Conflicts Of Interest ◽  
Lymphocytic Leukemia ◽  
Cpg Odn ◽  
Toll Like Receptor ◽  
Activated T Cells ◽  
Cpg Oligodeoxynucleotides ◽  
Interleukin 21 ◽  
Stimulation Of

Abstract Abstract 3685 Poster Board III-621 Interleukin 21 (IL-21) and CpG oligodeoxynucleotides (CpG ODN) are two novel and highly promising agents for the treatment of hematological diseases. Recently, we reported that IL-21 and CpG ODN induce granzyme B (GrB) and GrB-dependent apoptosis in malignant B cells from patients with B chronic lymphocytic leukemia (B-CLL), but not in healthy peripheral B cells. Using various techniques including FACS analysis, Western immunoblotting and RT-PCR we further characterized the factors accountable for the different apoptotic response of B-CLL cells versus normal B cells. GrB induction in B-CLL cells after stimulation with IL-21 and CpG ODN was associated with upregulation of transcription factors, which are normally involved in the differentiation of cytotoxic T lymphocytes (CTL) including T-bet, Eomesodermin (EOMES) and nuclear factor of activated T cells (NFAT), a finding not observed in normal healthy B cells. Furthermore, the induction of GrB in B-CLL cells by IL-21 and CpG ODN required signaling via a JAK/STAT-dependent pathway, as suggested by simultaneous upregulation of phosphorylated STAT3 and complete abrogation of GrB expression by the pan-JAK inhibitor pyridone 6. Stimulation of B-CLL cells with IL-21 and CpG ODN upregulated molecules involved in cell adhesion (CD54), antigen presentation (MHC class I), co-stimulation (CD40, CD86), and GrB uptake (CD222), suggesting B-CLL cells activated with IL-21 and CpG ODN are able to contact other immune cells and may be able to reabsorb secreted GrB. Similar findings resulted when the toll-like receptor (TLR)7 agonist imiquimod was used instead of the TLR9 agonist CpG ODN, suggesting that comparable differentiation programs are initiated by TLR7 and TLR9. In summary, B-CLL cells can express transcription factors involved in cytotoxic differentiation of CTL as well as GrB in response to IL-21 and TLR stimulation. The B-CLL cell differentiation program described in this study could explain our recent findings of GrB-dependent apoptosis in B-CLL cells but not in benign B cells. Moreover, our data provide novel insights into the aberrant signaling state of B-CLL cells. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Toll-Like Receptor 9: A New Target of Ischemic Preconditioning in the Brain

2008 ◽  
Vol 28 (5) ◽  
pp. 1040-1047 ◽  
Author(s):  
Susan L Stevens ◽  
Thomas MP Ciesielski ◽  
Brenda J Marsh ◽  
Tao Yang ◽  
Delfina S Homen ◽  
...  
Keyword(s):  
Brain Injury ◽  
Ischemic Injury ◽  
Ischemic Brain Injury ◽  
Dependent Manner ◽  
Cpg Odn ◽  
Toll Like Receptor ◽  
Ischemic Brain ◽  
Cpg Oligodeoxynucleotide ◽  
Tlr9 Ligand

Preconditioning with lipopolysaccharide (LPS), a toll-like receptor 4 (TLR4) ligand, provides neuroprotection against subsequent cerebral ischemic brain injury, through a tumor necrosis factor (TNF)α-dependent process. Here, we report the first evidence that another TLR, TLR9, can induce neuroprotection. We show that the TLR9 ligand CpG oligodeoxynucleotide (ODN) can serve as a potent preconditioning stimulus and provide protection against ischemic brain injury. Our studies show that systemic administration of CpG ODN 1826 in advance of brain ischemia (middle cerebral artery occlusion (MCAO)) reduces ischemic damage up to 60% in a dose- and time-dependent manner. We also offer evidence that CpG ODN preconditioning can provide direct protection to cells of the central nervous system, as we have found marked neuroprotection in modeled ischemia in vitro. Finally, we show that CpG preconditioning significantly increases serum TNFα levels before MCAO and that TNFα is required for subsequent reduction in damage, as mice lacking TNFα are not protected against ischemic injury by CpG preconditioning. Our studies show that preconditioning with a TLR9 ligand induces neuroprotection against ischemic injury through a mechanism that shares common elements with LPS preconditioning via TLR4.

scholarly journals Immunostimulatory CpG Oligodeoxynucleotide Confers Protection in a Murine Model of Infection with Burkholderia pseudomallei

2004 ◽  
Vol 72 (8) ◽  
pp. 4494-4502 ◽  
Author(s):  
Surasakdi Wongratanacheewin ◽  
Wannapa Kespichayawattana ◽  
Pakamas Intachote ◽  
Sathit Pichyangkul ◽  
Rasana W. Sermswan ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Burkholderia Pseudomallei ◽  
Bacterial Load ◽  
Elevated Serum ◽  
Interleukin 12 ◽  
Cpg Odn ◽  
Bacterial Challenge ◽  
Cpg Oligodeoxynucleotides ◽  
Cpg Oligodeoxynucleotide ◽  
Ifn Γ

ABSTRACT Although CpG oligodeoxynucleotides (CpG ODNs) are known to enhance resistance against infection in a number of animal models, little is known about the CpG-induced protection against acute fatal sepsis such as that associated with the highly virulent bacterium Burkholderia pseudomallei. We previously demonstrated in an in vitro study that immunostimulatory CpG ODN 1826 enhances phagocytosis of B. pseudomallei and induces nitric oxide synthase and nitric oxide production by mouse macrophages. In the present study, CpG ODN 1826 given intramuscularly to BALB/c mice 2 to 10 days prior to B. pseudomallei challenge conferred better than 90% protection. CpG ODN 1826 given 2 days before the bacterial challenge rapidly enhanced the innate immunity of these animals, judging from the elevated serum levels of interleukin-12 (IL-12)p70 and gamma interferon (IFN-γ) over the baseline values. No bacteremia was detected on day 2 in 85 to 90% of the CpG-treated animals, whereas more than 80% of the untreated animals exhibited heavy bacterial loads. Although marked elevation of IFN-γ was found consistently in the infected animals 2 days after the bacterial challenge, it was ameliorated by the CpG ODN 1826 pretreatment (P = 0.0002). Taken together, the kinetics of bacteremia and cytokine profiles presented are compatible with the possibility that protection by CpG ODN 1826 against acute fatal septicemic melioidosis in this animal model is associated with a reduction of bacterial load and interference with the potential detrimental effect of the robust production of proinflammatory cytokines associated with B. pseudomallei multiplication.

scholarly journals Free Feeding of CpG-Oligodeoxynucleotide Particles Prophylactically Attenuates Allergic Airway Inflammation and Hyperresponsiveness in Mice

2021 ◽  
Vol 12 ◽  
Author(s):  
Takuma Okajima ◽  
Suguru Shigemori ◽  
Fu Namai ◽  
Tasuku Ogita ◽  
Takashi Sato ◽  
...  
Keyword(s):  
Airway Inflammation ◽  
Oral Delivery ◽  
Allergic Airway Inflammation ◽  
Fecal Microbiota ◽  
Attractive Alternative ◽  
Cell Hyperplasia ◽  
Cpg Oligodeoxynucleotides ◽  
Cpg Oligodeoxynucleotide ◽  
Cpg Odns ◽  
Free Feeding

CpG-oligodeoxynucleotides (CpG-ODNs) constitute an attractive alternative for asthma treatment. However, very little evidence is available from studies on the oral administration of CpG-ODNs in animals. Previously, we developed acid-resistant particles (named ODNcap) as an oral delivery device for ODNs. Here, we showed that free feeding of an ODNcap-containing feed prophylactically attenuates allergic airway inflammation, hyperresponsiveness, and goblet cell hyperplasia in an ovalbumin-induced asthma model. Using transcriptomics-driven approaches, we demonstrated that injury of pulmonary vein cardiomyocytes accompanies allergen inhalation challenge, but is inhibited by ODNcap feeding. We also showed the participation of an airway antimicrobial peptide (Reg3γ) and fecal microbiota in the ODNcap-mediated effects. Collectively, our findings suggest that daily oral ingestion of ODNcap may provide preventive effects on allergic bronchopulmonary insults via regulation of mechanisms involved in the gut-lung connection.

scholarly journals Phosphate-modified CpG oligonucleotides induce in vitro maturation of human myeloid dendritic cells

2020 ◽  
Vol 24 (6) ◽  
pp. 653-660
Author(s):  
A. A. Ostanin ◽  
O. Y. Leplina ◽  
E. A. Burakova ◽  
T. V. Tyrinova ◽  
A. A. Fokina ◽  
...  
Keyword(s):  
Dendritic Cells ◽  
Cpg Odn ◽  
Blood Monocytes ◽  
Myeloid Dendritic Cells ◽  
Gm Csf ◽  
Cpg Oligodeoxynucleotides ◽  
Cpg Oligonucleotides ◽  
Cpg Odns ◽  
Dc Maturation

Myeloid dendritic cells (DCs) play an important role in the immune response; therefore, the search for compounds that can effectively activate DCs is a needful goal. This study was aimed to investigate the effect of synthetic CpG oligodeoxynucleotides (CpG-ODN) on the maturation and allostimulatory activity of myeloid DCs in comparison with other PAMP and DAMP molecules. For the research, we synthesized known CpG-ODN class C (SD-101 and D-SL03) containing thiophosphate internucleotide groups, and their original phosphate-modified analogues (SD-101M and D-SL03M) with mesylphosphoramide internucleotide groups (M = μ-modification). The effects of CpG-ODN and other activators were evaluated on DCs generated from blood monocytes in the presence of GM-CSF and IFN-α (IFN-DC) or IL-4 (IL4-DC). Evaluation of the intracellular TLR-9 expression showed that both types of DCs (IFN-DC and IL4-DC) contained on average 52 and 80 % of TLR-9-positive cells, respectively. The CpG-ODNs studied enhanced the allostimulatory activity of IFN-DCs, and the effect of μ-modified CpG-ODNs was higher than that of CpG-ODNs with thiophosphate groups. The stimulating effect of CpG-ODN at a dose of 1.0 μg/ml was comparable (for D-SL03, D-SL03M, SD-101) with or exceeded (for SD-101M) the effect of LPS at a dose of 10 μg/ml. At the same time, IFN-DCs were characterized by greater sensitivity to the action of CpG-ODNs than IL4-DCs. The enhancement of DC allostimulatory activity in the presence of CpG-ODNs was associated with the induction of final DC maturation, which was confirmed by a significant decrease in the number of CD14+DC, an increase in mature CD83+DC and a trend towards an increase in CD86+DC. Interestingly, the characteristic ability of LPS to enhance the expression of the co-stimulatory molecule OX40L on DCs was revealed only for the μ-analogue SD-101M. In addition, CpG-ODNs (SD-101 and SD-101M) had a stimulatory effect on IFN-γ production comparable to the action of LPS. The data obtained indicate a stimulating effect of CpG-ODN on the maturation and allostimulatory activity of human myeloid DCs, which is more pronounced for μ-modified analogs.

scholarly journals CpG Oligodeoxynucleotides for Anticancer Monotherapy from Preclinical Stages to Clinical Trials

Pharmaceutics ◽  
2021 ◽  
Vol 14 (1) ◽  
pp. 73
Author(s):  
Zhongkun Zhang ◽  
Jimmy Chun-Tien Kuo ◽  
Siyu Yao ◽  
Chi Zhang ◽  
Hira Khan ◽  
...  
Keyword(s):  
Clinical Trials ◽  
Anticancer Agents ◽  
Antitumor Immunity ◽  
Clinical Investigation ◽  
Cpg Odn ◽  
Tlr Agonists ◽  
Cpg Oligodeoxynucleotides ◽  
Cpg Odns ◽  
Future Direction ◽  
Tlr9 Activation

CpG oligodeoxynucleotides (CpG ODNs), the artificial versions of unmethylated CpG motifs that were originally discovered in bacterial DNA, are demonstrated not only as potent immunoadjuvants but also as anticancer agents by triggering toll-like receptor 9 (TLR9) activation in immune cells. TLR9 activation triggered by CpG ODN has been shown to activate plasmacytoid dendritic cells (pDCs) and cytotoxic T lymphocytes (CTLs), enhancing T cell-mediated antitumor immunity. However, the extent of antitumor immunity carried by TLR agonists has not been optimized individually or in combinations with cancer vaccines, resulting in a decreased preference for TLR agonists as adjuvants in clinical trials. Although various combination therapies involving CpG ODNs have been applied in clinical trials, none of the CpG ODN-based drugs have been approved by the FDA, owing to the short half-life of CpG ODNs in serum that leads to low activation of natural killer cells (NK cells) and CTLs, along with increases of pro-inflammatory cytokine productions. This review summarized the current innovation on CpG ODNs that are under clinical investigation and explored the future direction for CpG ODN-based nanomedicine as an anticancer monotherapy.

scholarly journals Formulation of bovine respiratory syncytial virus fusion protein with CpG oligodeoxynucleotide, cationic host defence peptide and polyphosphazene enhances humoral and cellular responses and induces a protective type 1 immune response in mice

2009 ◽  
Vol 90 (8) ◽  
pp. 1892-1905 ◽  
Author(s):  
J. Kovacs-Nolan ◽  
J. W. Mapletoft ◽  
Z. Lawman ◽  
L. A. Babiuk ◽  
S. van Drunen Littel-van den Hurk
Keyword(s):  
Immune Response ◽  
Respiratory Syncytial Virus ◽  
Immune Responses ◽  
Fusion Protein ◽  
Vaccine Development ◽  
Host Defence ◽  
Cpg Odn ◽  
Cpg Oligodeoxynucleotides ◽  
Cpg Oligodeoxynucleotide ◽  
Syncytial Virus

Respiratory syncytial virus (RSV) is the leading cause of serious respiratory tract disease in children and calves; however, RSV vaccine development has been slow due to early observations that formalin-inactivated vaccines induced Th2-type immune responses and led to disease enhancement upon subsequent exposure. Hence, there is a need for novel adjuvants that will promote a protective Th1-type or balanced immune response against RSV. CpG oligodeoxynucleotides (ODNs), indolicidin, and polyphosphazene were examined for their ability to enhance antigen-specific immune responses and influence the Th-bias when co-formulated with a recombinant truncated bovine RSV (BRSV) fusion protein (ΔF). Mice immunized with ΔF co-formulated with CpG ODN, indolicidin, and polyphosphazene (ΔF/CpG/indol/PP) developed higher levels of ΔF-specific serum IgG, IgG1 and IgG2a antibodies when compared with ΔF alone, and displayed an increase in the frequency of gamma interferon-secreting cells and decreased interleukin (IL)-5 production by in vitro restimulated splenocytes, characteristic of a Th1 immune response. These results were observed in both C57BL/6 and BALB/c strains of mice. When evaluated in a BRSV challenge model, mice immunized with ΔF/CpG/indol/PP developed significantly higher levels of BRSV-neutralizing serum antibodies than mice immunized with the ΔF protein alone, and displayed significantly less pulmonary IL-4, IL-5, IL-13 and eotaxin and reduced eosinophilia after challenge. These results suggest that co-formulation of ΔF with CpG ODN, host defence peptide and polyphosphazene may result in a safe and effective vaccine for the prevention of BRSV and may have implications for the development of novel human RSV vaccines.

scholarly journals CpG Oligodeoxynucleotide Developed For Activating Primate Immune Responses Promotes Anti-Tumoral Effect Combine With Neo-Antigen Based mRNA Cancer Vaccine

2021 ◽  
Author(s):  
Rongkuan Hu ◽  
Qin Li ◽  
Jie Ren ◽  
Wei Liu ◽  
Guoqin Jiang
Keyword(s):  
Immune Responses ◽  
Cancer Vaccine ◽  
Cancer Vaccines ◽  
Cytokine Production ◽  
Toll Like Receptor ◽  
Human Pbmc ◽  
Cpg Oligodeoxynucleotides ◽  
Adaptive Immune ◽  
Cpg Oligodeoxynucleotide

Abstract Synthetic phosphorthiolate modified CPG-oligodeoxynucleotides (CPG-ODN) activate innate and adaptive immune responses, which being exploited as a therapeutic approach. Here, we first screened and identified a new CpG-B class ODN (CpG2018B) that effectively stimulates type II interferon both in mouse Plasmacytoid dendritic cells (p-DC) and human PBMC. In addition, CpG2018B promotes cytokine production mainly via toll-like receptor 9 (TLR9) pathways. We further demonstrated that intratumoral (IT) injection of CpG2018B inhibits melanoma growth in syngeneic models and could turn “cold” tumors into “hot” tumors. Then, CpG2018B and mRNA based neo-antigen cancer vaccine were encapsulated into lipid-nanoparticle (LNP) and intratumoral injected into melanoma mice models. Interestingly, vaccination with CpG or mRNA vaccine alone could inhibit tumor growth respectively, while, CpG combine with mRNA vaccine enhanced the anti-tumor effect. At last, we described the long-term safety and tolerability of CpG2018B and mRNA therapy in mice models. In conclusion, we identified a novel CpG-B ODNs to promote immune response and CpG combine with mRNA cancer vaccines are attractive candidate for immune stimulatory sequences (ISS) based therapeutic strategies.

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