scholarly journals Development of Nonaggregating Poly-A Tailed Immunostimulatory A/D Type CpG Oligodeoxynucleotides Applicable for Clinical Use

2015 ◽  
Vol 2015 ◽  
pp. 1-20 ◽  
Author(s):  
Taiki Aoshi ◽  
Yasunari Haseda ◽  
Kouji Kobiyama ◽  
Hirotaka Narita ◽  
Hideaki Sato ◽  
...  
Keyword(s):  
Physical Property ◽  
Good Manufacturing Practice ◽  
Vaccine Adjuvant ◽  
Drug Preparation ◽  
Dependent Manner ◽  
Human Pbmcs ◽  
Vaccine Adjuvants ◽  
Immunostimulatory Activity ◽  
Cpg Oligodeoxynucleotides ◽  
Cpg Odns

Immunostimulatory CpG ODNs have been developed and utilized as TLR9-dependent innate immune activators and vaccine adjuvants. Four different types of immunostimulatory CpG ODNs (A/D, B/K, C, and P type) have been reported. A/D type ODNs are characterized by high IFN-αproduction but intrinsically form aggregates, hindering its good manufacturing practice grade preparation. In this study, we developed several D35-derived ODNs (a commonly used A/D type ODN), which were modified with the addition of a phosphorothioate polynucleotide tail (such as dAs40), and examined their physical properties, solubility in saline, immunostimulatory activity on human PBMCs, and vaccine adjuvant potential in monkeys. We found that two modified ODNs including D35-dAs40 and D35core-dAs40 were immunostimulatory, similar to original D35 in human PBMCs, resulting in high IFN-αsecretion in a dose-dependent manner. Physical property analysis by dynamic light scattering revealed that both D35-dAs40 and D35core-dAs40 did not form aggregates in saline, which is currently impossible for the original D35. Furthermore, D35-dAs40 and D35core-dAs40 worked as better vaccine adjuvant in monkeys. These results suggested that D35-dAs40 and D35core-dAs40 are two promising prototypes of nonaggregating A/D type ODN with advantages of ease of drug preparation for clinical applications as vaccine adjuvants or IFN-αinducing immunomodifiers.

scholarly journals Immunomodulatory Effects of CpG Oligodeoxynucleotides on Established Th2 Responses

2002 ◽  
Vol 9 (6) ◽  
pp. 1260-1269 ◽  
Author(s):  
Kunihiko Kitagaki ◽  
Vipul V. Jain ◽  
Thomas R. Businga ◽  
Iftikhar Hussain ◽  
Joel N. Kline
Keyword(s):  
Allergic Diseases ◽  
Interleukin 12 ◽  
Dependent Manner ◽  
Independent Manner ◽  
Concentration Dependent Manner ◽  
Cpg Oligodeoxynucleotides ◽  
Th2 Responses ◽  
Cpg Odns ◽  
Ifn Γ

ABSTRACT CpG oligodeoxynucleotides (CpG ODNs) are known to induce type 1 T-helper-cell (Th1) responses. We have previously demonstrated that CpG ODNs administered during sensitization prevent Th2-mediated eosinophilic airway inflammation in vivo. We also reported that key Th1 cytokines, gamma interferon (IFN-γ) and interleukin 12 (IL-12), are not necessary for this protection. Recent in vivo data suggest that CpG ODNs might also reverse established pulmonary eosinophilia. In order to clarify how CpG ODNs can inhibit established Th2 responses, we evaluated the cytokine production from splenocytes from antigen- and alum-immunized mice. Restimulation with antigen induced IL-5, which was clearly inhibited by coculture with CpG ODNs in a concentration-dependent manner. CpG ODNs also induced IFN-γ, but in a concentration-independent manner. The inhibition of IL-5 production was not mediated through natural killer cells or via CD8+ T lymphocytes. Although IFN-γ plays an important role in inhibition of antigen-induced IL-5 production by CpG ODNs, IFN-γ was not the sole factor in IL-5 inhibition. CpG ODNs also induced IL-10, and this induction correlated well with IL-5 inhibition. Elimination of IL-10 reduced the anti-IL-5 effect of CpG ODNs, although incompletely. This may be because IFN-γ, induced by CpG ODNs, is also inhibited by IL-10, serving as a homeostatic mechanism for the Th1-Th2 balance. Overproduction of IFN-γ was downregulated by CpG ODN-induced IL-10 via modulation of IL-12 production. These data suggest that CpG ODNs may inhibit established Th2 immune responses through IFN-γ and IL-10 production, the latter serving to regulate excessive Th1 bias. These properties of CpG ODNs might be a useful feature in the development of immunotherapy adjuvants against allergic diseases such as asthma.

scholarly journals Vaccination with Heat-killed Leishmania Antigen or Recombinant Leishmanial Protein and CpG Oligodeoxynucleotides Induces Long-Term Memory CD4+and CD8+T Cell Responses and Protection Against Leishmania major Infection

2002 ◽  
Vol 195 (12) ◽  
pp. 1565-1573 ◽  
Author(s):  
Elizabeth G. Rhee ◽  
Susana Mendez ◽  
Javeed A. Shah ◽  
Chang-you Wu ◽  
Joanna R. Kirman ◽  
...  
Keyword(s):  
T Cells ◽  
Cd8 T Cells ◽  
Leishmania Major ◽  
Vaccine Adjuvant ◽  
Dependent Manner ◽  
Cpg Odn ◽  
Major Infection ◽  
Cpg Oligodeoxynucleotides ◽  
Immune Correlates

CpG oligodeoxynucleotides (ODN) have potent effects on innate and adaptive cellular immune responses. In this report, the ability of CpG ODN to confer long-term immunity and protection when used as a vaccine adjuvant with a clinical grade of leishmanial antigen, autoclaved Leishmania major (ALM), or a recombinant leishmanial protein was studied. In two different mouse models of L. major infection, vaccination with ALM plus CpG ODN was able to control infection and markedly reduce lesion development in susceptible BALB/c and resistant C57BL/6 (B6) mice, respectively, up to 12 wk after immunization. Moreover, B6 mice immunized with ALM plus CpG ODNs were still protected against infectious challenge even 6 mo after vaccination. In terms of immune correlates of protection, ALM plus CpG ODN-vaccinated mice displayed L. major–specific T helper cell 1 and CD8+ responses. In addition, complete protection was markedly abrogated in mice depleted of CD8+ T cells at the time of vaccination. Similarly, mice vaccinated with a recombinant leishmanial protein plus CpG ODN also had long-term protection that was dependent on CD8+ T cells in vivo. Together, these data demonstrate that CpG ODN, when used as a vaccine adjuvant with either a recombinant protein or heat-killed leishmanial antigen, can induce long-term protection against an intracellular infection in a CD8-dependent manner.

scholarly journals Enhancement of the Immunostimulatory Effect of Phosphodiester CpG Oligodeoxynucleotides by an Antiparallel Guanine-Quadruplex Structural Scaffold

Biomolecules ◽  
2021 ◽  
Vol 11 (11) ◽  
pp. 1617
Author(s):  
Fika Ayu Safitri ◽  
Anh Thi Tram Tu ◽  
Kazuaki Hoshi ◽  
Miwako Shobo ◽  
Dandan Zhao ◽  
...  
Keyword(s):  
Rational Design ◽  
Mrna Levels ◽  
Mouse Macrophage ◽  
Flanking Sequence ◽  
Immunostimulatory Activity ◽  
Cpg Oligodeoxynucleotides ◽  
Cpg Motifs ◽  
Guanine Quadruplex ◽  
Cpg Odns ◽  
High Level

Guanine-quadruplex-based CpG oligodeoxynucleotides (G4 CpG ODNs) have been developed as potent immunostimulatory agents with reduced sensitivity to nucleases. We designed new monomeric G4 ODNs with an antiparallel topology using antiparallel type duplex/G4 ODNs as robust scaffolds, and we characterized their topology and effects on cytokine secretion. Based on circular dichroism analysis and quantification of mRNA levels of immunostimulatory cytokines, it was found that monomeric antiparallel G4 CpG ODNs containing two CpG motifs in the first functional loop, named G2.0.0, could maintain antiparallel topology and generate a high level of immunostimulatory cytokines in RAW264 mouse macrophage-like cell lines. We also found that the flanking sequence in the CpG motif altered the immunostimulatory effects. Gc2c.0.0 and Ga2c.0.0 are monomeric antiparallel G4 CpG ODNs with one cytosine in the 3′ terminal and one cytosine/adenine in the 5′ terminal of CpG motifs that maintained the same resistance to degradation in serum as G2.0.0 and improved interleukin-6 production in RAW264 and bone marrow-derived macrophages. The immunostimulatory activity of antiparallel G4 CpG ODNs is superior to that of linear natural CpG ODNs. These results provide insights for the rational design of highly potent CpG ODNs using antiparallel G4 as a robust scaffold.

The Application and Functional Progress of γ-Poly-Glutamic Acid in Food: A Mini-Review

2020 ◽  
Vol 26 (41) ◽  
pp. 5347-5352
Author(s):  
Guoliang Wang ◽  
Qing Liu ◽  
Ying Wang ◽  
Jingyuan Li ◽  
Yue Chen ◽  
...  
Keyword(s):  
Glutamic Acid ◽  
Food Industry ◽  
Water Solubility ◽  
Drug Carrier ◽  
Physical Property ◽  
Coating Material ◽  
Vaccine Adjuvant ◽  
Food Ingredient ◽  
Naturally Occurring

γ-Poly-glutamic acid (γ-PGA) is a naturally occurring homo-polyamide produced by various strains of Bacillus. As a biopolymer substance, γ-PGA possesses a few predominant features containing good water solubility, biocompatibility, degradability and non-toxicity. Based on this, γ-PGA can be used in pharmaceutical, such as drug carrier/deliverer, vaccine adjuvant, and coating material for microencapsulation, etc. Moreover, it has also been applied in a broad range of industrial fields including food, medicine, bioremediation, cosmetics, and agriculture. Especially, γ-PGA is an extremely promising food ingredient. In this mini-review, our aim is to review the function and application progress of γ-PGA in the food industry: e.g., improving taste and flavor, enhancing physical property, and promoting health.

scholarly journals Magic Peptide: Unique Properties of the LRR11 Peptide in the Activation of Leukotriene Synthesis in Human Neutrophils

2021 ◽  
Vol 22 (5) ◽  
pp. 2671
Author(s):  
Galina M. Viryasova ◽  
Ekaterina A. Golenkina ◽  
Tibor Hianik ◽  
Nataliya V. Soshnikova ◽  
Nina G. Dolinnaya ◽  
...  
Keyword(s):  
Defense Mechanisms ◽  
Specific Binding ◽  
Acoustic Method ◽  
Effective Vaccine ◽  
Human Neutrophils ◽  
Shear Mode ◽  
Vaccine Adjuvants ◽  
High Biological Activity ◽  
Pathogen Elimination ◽  
Cpg Odns

Neutrophil-mediated innate host defense mechanisms include pathogen elimination through bacterial phagocytosis, which activates the 5-lipoxygenase (5-LOX) product synthesis. Here, we studied the effect of synthetic oligodeoxyribonucleotides (ODNs), which mimic the receptor-recognized sites of bacterial (CpG-ODNs) and genomic (G-rich ODNs) DNAs released from the inflammatory area, on the neutrophil functions after cell stimulation with Salmonella typhimurium. A possible mechanism for ODN recognition by Toll-like receptor 9 (TLR9) and RAGE receptor has been proposed. We found for the first time that the combination of the magic peptide LRR11 from the leucine-rich repeat (LRR) of TLR9 with the CpG-ODNs modulates the uptake and signaling from ODNs, in particular, dramatically stimulates 5-LOX pathway. Using thickness shear mode acoustic method, we confirmed the specific binding of CpG-ODNs, but not G-rich ODN, to LRR11. The RAGE receptor has been shown to play an important role in promoting ODN uptake. Thus, FPS-ZM1, a high-affinity RAGE inhibitor, suppresses the synthesis of 5-LOX products and reduces the uptake of ODNs by neutrophils; the inhibitor effect being abolished by the addition of LRR11. The results obtained revealed that the studied peptide-ODN complexes possess high biological activity and can be promising for the development of effective vaccine adjuvants and antimicrobial therapeutics.

scholarly journals Adjuvanticity of a synthetic cord factor analogue for subunit Mycobacterium tuberculosis vaccination requires FcRγ–Syk–Card9–dependent innate immune activation

2009 ◽  
Vol 206 (1) ◽  
pp. 89-97 ◽  
Author(s):  
Kerstin Werninghaus ◽  
Anna Babiak ◽  
Olaf Groß ◽  
Christoph Hölscher ◽  
Harald Dietrich ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Subunit Vaccine ◽  
Vaccine Development ◽  
Adaptor Protein ◽  
Synthetic Analogue ◽  
Dependent Manner ◽  
Immunostimulatory Activity ◽  
Adjuvant Activity ◽  
Cord Factor

Novel vaccination strategies against Mycobacterium tuberculosis (MTB) are urgently needed. The use of recombinant MTB antigens as subunit vaccines is a promising approach, but requires adjuvants that activate antigen-presenting cells (APCs) for elicitation of protective immunity. The mycobacterial cord factor Trehalose-6,6-dimycolate (TDM) and its synthetic analogue Trehalose-6,6-dibehenate (TDB) are effective adjuvants in combination with MTB subunit vaccine candidates in mice. However, it is unknown which signaling pathways they engage in APCs and how these pathways are coupled to the adaptive immune response. Here, we demonstrate that these glycolipids activate macrophages and dendritic cells (DCs) via Syk–Card9–Bcl10–Malt1 signaling to induce a specific innate activation program distinct from the response to Toll-like receptor (TLR) ligands. APC activation by TDB and TDM was independent of the C-type lectin receptor Dectin-1, but required the immunoreceptor tyrosine-based activation motif–bearing adaptor protein Fc receptor γ chain (FcRγ). In vivo, TDB and TDM adjuvant activity induced robust combined T helper (Th)-1 and Th-17 T cell responses to a MTB subunit vaccine and partial protection against MTB challenge in a Card9-dependent manner. These data provide a molecular basis for the immunostimulatory activity of TDB and TDM and identify the Syk–Card9 pathway as a rational target for vaccine development against tuberculosis.

scholarly journals Inflammatory arthritis can be reined in by CpG-induced DC–NK cell cross talk

2007 ◽  
Vol 204 (8) ◽  
pp. 1911-1922 ◽  
Author(s):  
Hsin-Jung Wu ◽  
Heloisa Sawaya ◽  
Bryce Binstadt ◽  
Margot Brickelmaier ◽  
Amanda Blasius ◽  
...  
Keyword(s):  
Immune System ◽  
Cross Talk ◽  
Inflammatory Arthritis ◽  
Nk Cell ◽  
Therapeutic Potential ◽  
Adaptive Immune System ◽  
Interleukin 12 ◽  
Inhibitory Influence ◽  
Cpg Oligodeoxynucleotides ◽  
Cpg Odns

Unmethylated CpG-oligodeoxynucleotides (ODNs) are generally thought of as potent adjuvants with considerable therapeutic potential to enhance immune responses against microbes and tumors. Surprisingly, certain so-called stimulatory CpG-ODNs strongly inhibited the effector phase of inflammatory arthritis in the K/BxN serum transfer system, either preventively or therapeutically. Also unexpected was that the inhibitory influence did not depend on the adaptive immune system cells mobilized in an immunostimulatory context. Instead, they relied on cells of the innate immune system, specifically on cross talk between CD8α+ dendritic cells and natural killer cells, resulting in suppression of neutrophil recruitment to the joint, orchestrated through interleukin-12 and interferon-γ. These findings highlight potential applications of CpG-ODNs and downstream molecules as antiinflammatory agents.

scholarly journals Antitumor and Immunostimulatory Activity of Two Chromones and Other Constituents from Cassia Petersiana

2006 ◽  
Vol 1 (11) ◽  
pp. 1934578X0600101 ◽  
Author(s):  
Pierre C. Djemgou ◽  
Donatien Gatsing ◽  
Marguérite Tchuendem ◽  
Bonaventure T. Ngadjui ◽  
Pierre Tane ◽  
...  
Keyword(s):  
Antioxidant Activities ◽  
Raw 264.7 Cells ◽  
Proliferation Index ◽  
Dependent Manner ◽  
Hep G2 ◽  
Immunostimulatory Activity ◽  
Nmr Studies ◽  
Biological Investigation ◽  
Dose Dependent ◽  
Mcf 7

Phytochemical and biological investigation of the leaves of Cassia petersiana afforded two new chromones (1, 2), in addition, to the known glyceryl-1-hexacosanoate (3) and stigmasterol-3-O-β-D-glucoside (4). The structures of the compounds were determined by comprehensive NMR studies, including DEPT, COSY, HMQC, HMBC and IR spectroscopy and MS. 1–4 were investigated against different types of cell lines, including solid tumor cells (Hep-G2, and MCF-7 cells) and leukemia (1301) cells for their cytotoxic effects. 1–3 possessed a dose-dependent cytotoxic effect against Hep-G2 cells, but in relatively high concentrations; 4 was the most cytotoxic with the lowest IC50 value of 82.7 μM. The calculated IC50 values against MCF-7 cells were 112.2 μM, 143.7 μM, 68.1 μM, and 114.3 μM for 1, 2, 3, and 4, respectively. The alteration in the macrophage proliferation index, using Raw 264.7 cells, was monitored. 1 and 3 were the highest stimulators of macrophage proliferation in a dose-dependent manner, whereas 2 and 4 showed a peak point of stimulation at 20 μM. The effect of these compounds on pre-induced NO was explored. 1–4 inhibited the LPS-induced NO, with inhibition percentages of 80.5%, 89.3%, 82.1%, and 92.1%, respectively, at a concentration of 20 μM. The antioxidant capacity of 1–4, using the DPPH assay was also investigated. 1–3 possessed weak scavenging activity; while 4 had an effective SC50 value as low as 36 μM. These results indicated that 4 possessed the highest anti-tumor, immunoproliferative, macrophage proliferation, anti-inflammatory, and antioxidant activities.

scholarly journals PMA Induces Vaccine Adjuvant Activity by the Modulation of TLR Signaling Pathway

2014 ◽  
Vol 2014 ◽  
pp. 1-8 ◽  
Author(s):  
Dool-Ri Oh ◽  
Hu Won Kang ◽  
Jong-Ro Kim ◽  
Sunoh Kim ◽  
In-Kyu Park ◽  
...  
Keyword(s):  
Active Component ◽  
Mucosal Vaccine ◽  
Vaccine Adjuvant ◽  
Vaccine Adjuvants ◽  
Adjuvant Activity ◽  
Independent Manner ◽  
Etoh Extract ◽  
Croton Tiglium ◽  
Fractionation Column

Toll-like receptor (TLR) ligands are being developed for use as vaccine adjuvants and as immunomodulators because of their ability to stimulate innate and adaptive immune responses. Flagellin, a TLR5 ligand, was reported to show potent mucosal vaccine adjuvant activity. To identify ligands that potentiate the adjuvant activity of flagellin, we screened a plant library using HEK293T cells transiently cotransfected with phTLR5 and pNF-κB-SEAP plasmids. The 90% EtOH extract fromCroton tigliumshowed significant NF-κB transactivation in a TLR5-independent manner along with the increase of a flagellin activity. We have studied to characterize an active component fromCroton tigliumand to elucidate the action mechanisms. Phorbol 12-myristate 13-acetate (PMA) was isolated as an active component ofCroton tigliumby activity-guided fractionation, column chromatography, HPLC, NMR, and MS. PMA at a range of nM induced PKC-dependent NF-κB activation and IL-8 production in both TLR5− and TLR5+ assay systems. In in vivo mouse vaccination model, PMA induced antigen-specific IgG and IgA antibody responses and increased IL-12 production corresponding to T cell responses in spleen lymphocytes. These results suggest that PMA would serve as an efficacious mucosal vaccine adjuvant.

scholarly journals CpG Oligodeoxynucleotides Enhance Host Defense during Murine Tuberculosis

2002 ◽  
Vol 70 (1) ◽  
pp. 147-152 ◽  
Author(s):  
Nicole P. Juffermans ◽  
Jaklien C. Leemans ◽  
Sandrine Florquin ◽  
Annelies Verbon ◽  
Arend H. Kolk ◽  
...  
Keyword(s):  
Pulmonary Tuberculosis ◽  
Host Defense ◽  
Interleukin 4 ◽  
T Helper 1 ◽  
Th1 Response ◽  
Cpg Oligodeoxynucleotides ◽  
Cpg Odns ◽  
Ifn Γ ◽  
Deficient Mice ◽  
Dependent Mechanism

ABSTRACT Oligodeoxynucleotides (ODNs) containing unmethylated CpG motifs activate immune cells to produce cytokines. CpG ODNs protect mice against infections with intracellular bacteria by the induction of a T helper 1 (Th1) response. To determine the effect of CpG ODNs in pulmonary tuberculosis, mice were treated with CpG ODNs or control ODNs at the time of intranasal infection. CpG ODNs reduced mycobacterial outgrowth for up to 5 weeks after Mycobacterium tuberculosis infection and were associated with a decrease in inflammation in lung tissue. CpG treatment was also associated with elevated levels of gamma interferon (IFN-γ) and decreased levels of interleukin 4 in the lungs and an increased capacity of splenocytes to secrete Th1-type cytokines. CpG ODNs given 2 weeks after infection were still able to reduce mycobacterial outgrowth and to enhance a Th1 response 5 weeks postinfection. Administration of CpG ODNs to IFN-γ-gene-deficient mice failed to reduce mycobacterial outgrowth. These data suggest that CpG ODNs improve host defense during pulmonary tuberculosis by an IFN-γ-dependent mechanism.

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