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Fish & Shellfish Immunology Dec 2017Poly (I:C) showed promise as an immunoprotective agents in rock bream against rock bream iridovirus (RBIV) infection. In this study, we evaluated the time-dependent...
Poly (I:C) showed promise as an immunoprotective agents in rock bream against rock bream iridovirus (RBIV) infection. In this study, we evaluated the time-dependent virus replication pattern and antiviral immune responses in RBIV-infected rock bream with and without poly (I:C) administration. In the poly (I:C)+virus-injected group, virus copy numbers were more than 18.9-, 24.0- and 479.2-fold lower than in the virus only injected group at 4 (4.73 × 10 and 8.95 × 10/μl, respectively), 7 (3.67 × 10 and 8.81 × 10/μl, respectively) and 10 days post infection (dpi) (1.26 × 10 and 6.02 × 10/μl, respectively). Moreover, significantly high expression levels of TLR3 (8.6- and 7.7-fold, at 4 and 7 dpi, respectively) and IL1β (3.6-fold at 2 dpi) were observed in the poly (I:C)+virus-injected group, but the expression levels were not significantly in the virus-injected group. However, IL8 and TNFα expression levels showed no statistical significance in both groups. Mx, ISG15 and PKR were significantly highly expressed from 4 to 10 dpi in the virus-injected group. Nevertheless, in the poly (I:C)+virus-injected group, Mx and ISG15 expression were significantly expressed from 2 dpi. In summary, poly (I:C) administration in rock bream induces TLR3, IL1β, Mx and ISG15-mediated immune responses, which could be a critical factor for inhibition of virus replication.
Topics: Animals; DNA Virus Infections; Fish Diseases; Immunity, Innate; Iridoviridae; Perciformes; Poly I-C; Virus Replication
PubMed: 28986216
DOI: 10.1016/j.fsi.2017.10.002 -
Molecular Immunology Oct 2016The Pacific oyster (Crassostrea gigas) is farmed globally. Ostreid herpesvirus (OsHV-1) causes severe mortalities of farmed C. gigas. Management of OsHV-1 has proven...
The Pacific oyster (Crassostrea gigas) is farmed globally. Ostreid herpesvirus (OsHV-1) causes severe mortalities of farmed C. gigas. Management of OsHV-1 has proven difficult. Oysters treated with poly(I:C) exhibit enhanced protection (EP) against OsHV-1. This chemical treatment is highly effective, but it is not feasible to treat every oyster on a farm. To circumvent this practical limitation, previous studies on arthropods have suggested that EP can be transferred from parents to their offspring (trans-generational EP, TGEP). This suggests that the treatment of relatively few parents could be used to produce large numbers of offspring with TGEP. Here, we investigated TGEP in oysters to test whether it might be used as a cost effective management tool to control OsHV-1. We found that offspring (D-veliger larvae) produced from poly(I:C)-treated parents had double the chance of surviving exposure to OsHV-1 compared to controls. Furthermore, the larvae of poly(I:C)-treated parents contained elevated levels of mRNA encoding a key transcription factor that regulates antiviral immunity (IRF2). Poly(I:C) treatment had no effect on the survival of oyster parents. Hence, the enhanced immunity of their offspring could not be explained by genetic selection, and instead may reflect epigenetic reprogramming or maternal provisioning.
Topics: Animals; Antiviral Agents; Crassostrea; Herpesviridae; Herpesviridae Infections; Poly I-C; Real-Time Polymerase Chain Reaction
PubMed: 27616590
DOI: 10.1016/j.molimm.2016.09.002 -
FEBS Letters Nov 2010Several RNA viruses can be detected by the inflammasome, which promotes IL-1β and IL-18 secretion, but the underlying mechanisms of detection remain unclear. Cytosolic...
Several RNA viruses can be detected by the inflammasome, which promotes IL-1β and IL-18 secretion, but the underlying mechanisms of detection remain unclear. Cytosolic dsRNA is a replication intermediate of many RNA viruses. We show here that transfection of the dsRNA analogue poly I:C activates the NLRP3 inflammasome via a pathway requiring endosomal acidification. This detection is independent of the other poly I:C sensors: TLR3 and MDA5. These results suggest a mechanism by which cytosolic dsRNA produced during viral infection could activate the NLRP3 inflammasome.
Topics: Animals; Carrier Proteins; DNA, Viral; Dendritic Cells; Electroporation; Endosomes; Female; Hydrogen-Ion Concentration; Inflammasomes; Interleukin-1beta; Intracellular Space; Ligands; Macrophages; Mice; NLR Family, Pyrin Domain-Containing 3 Protein; Poly I-C; Transfection
PubMed: 20971108
DOI: 10.1016/j.febslet.2010.10.036 -
Biomedicine / [publiee Pour... Oct 1977In several mouse strains, which exhibited variability in degree of immune responsiveness, concomitant i.p. injection of antigen (sheep erythrocytes) and poly I: poly C...
In several mouse strains, which exhibited variability in degree of immune responsiveness, concomitant i.p. injection of antigen (sheep erythrocytes) and poly I: poly C enhanced the early rate of increase in antibody plaque-forming cells (PFC) in the spleen. This adjuvant effect of poly I: poly C was most marked in LACA mice and was demonstrable over a range of antigen and polynucleotide concentrations. The double-stranded RNA did not however, affect the number of PFC at the peak of the primary response. Stimulation was dependent on the temporal relationship between antigen and adjuvant administration--injection of poly I: poly C 24 and 48 h prior to antigen causing suppression of humoral immunity. Poly I: Poly C had a progressively more marked enhancing effect on PFC production at high (thymus-independent) doses of erythrocytes. This suggests that its adjuvant action may be mediated via cells other than T cells. Administration of poly I: poly C caused an initial depression followed by stimulation of spleen weight and nucleated cell numbers in sensitized animals. The possible mechanism(s) of action of this polynucleotide adjuvant are discussed in the light of these and other findings.
Topics: Animals; Antibody-Producing Cells; Dose-Response Relationship, Drug; Erythrocytes; Hemolytic Plaque Technique; Kinetics; Male; Mice; Organ Size; Poly I-C; Sheep; Spleen; Stimulation, Chemical; Time Factors
PubMed: 334274
DOI: No ID Found -
International Immunology May 2004Dendritic cells (DC) are professional antigen-presenting cells that play a central role in the control of immunity. Mature DC are characterized by high expression levels...
Dendritic cells (DC) are professional antigen-presenting cells that play a central role in the control of immunity. Mature DC are characterized by high expression levels of MHC and co-stimulatory molecules, and by the secretion of IL-12, a key cytokine for the priming of cytotoxic T lymphocytes. Here, we have compared different maturation stimuli to reproducibly generate stable mature DC secreting high amounts of bioactive IL-12p70. We have compared soluble human trimeric CD40 ligand (sCD40L) combined with IFN-gamma, poly(I:C), a cocktail of cytokines (IL-1beta, IL-6 and tumor necrosis factor-alpha) with prostaglandin E(2) and lipopolysaccharide. A major concern, however, is whether DC, that have already produced high amounts of IL-12p70 during the maturation step, are still capable of secreting IL-12p70 after in vivo administration at the time of interaction with the targeted T cells. To mimic that situation, mature DC generated by those methods were compared for their ability to secrete IL-12p70 in the absence of IFN-gamma, using sCD40L. We observed a second consistent secretion of bioactive IL-12p70 upon subsequent sCD40L stimulation only when poly(I:C) was used as the maturating agent. Our data suggest that, for clinical use, poly(I:C) may be one of the most appropriate agents to generate stable mature DC. These mature DC might generate in vivo effective immune responses after injection, because they retain the ability to secrete bioactive IL-12 after CD40 ligation.
Topics: Antigens, Surface; Dendritic Cells; Humans; Immunotherapy; Interleukin-12; Poly I-C; Protein Subunits
PubMed: 15096480
DOI: 10.1093/intimm/dxh077 -
Journal of Biological Response Modifiers Dec 1985We have performed Phase I trials of two synthetic double-stranded polyribonucleotide complexes--poly(I,C)-LC, a complex of polyinosinic-polycytidylic acid with...
We have performed Phase I trials of two synthetic double-stranded polyribonucleotide complexes--poly(I,C)-LC, a complex of polyinosinic-polycytidylic acid with poly-L-lysine and carboxymethylcellulose, and poly(I,C)-L, which lacks carboxymethylcellulose--in patients with advanced cancer. With poly(I,C)-LC, several treatment schedules were investigated in an attempt to decrease toxicity and maximize interferon (IFN) induction. The best tolerated was an alternate-day schedule, with gradual dose escalation. Daily short infusions and continuous (24-h) infusions were tolerated less well. Maximum tolerated doses varied over a several hundredfold dose range. Toxicity consisted of fever, rigors, hypotension, and blood count depression. Two patients treated with poly(I,C)-L developed systemic allergic reactions, and antibodies to poly(I,C)-L and its components were detected in the serum of some patients treated with both compounds. IFN-alpha was induced in most patients at serum levels similar to those achieved after intramuscular administration of human IFN-alpha. Of 32 patients, one with renal cell carcinoma showed partial tumor regression. Poly(I,C) complexes are effective IFN inducers in humans, but their toxicity limits their use in cancer patients.
Topics: Adult; Aged; Antibody Formation; Carboxymethylcellulose Sodium; Carcinoma, Renal Cell; Drug Evaluation; Drug Tolerance; Female; Humans; Hypotension; Interferons; Kidney Neoplasms; Killer Cells, Natural; Leukopenia; Male; Methylcellulose; Middle Aged; Neoplasms; Poly I-C; Polylysine
PubMed: 2418162
DOI: No ID Found -
Journal of Controlled Release :... Mar 2011The combination of nucleic acid-based Toll-like receptor (TLR)-3 or TLR9 agonists and cationic liposomes constitutes an effective vaccine adjuvant approach for eliciting...
The combination of nucleic acid-based Toll-like receptor (TLR)-3 or TLR9 agonists and cationic liposomes constitutes an effective vaccine adjuvant approach for eliciting CD8+ T-cell responses. However, complexing cationic liposomes and oppositely charged oligonucleotides generally results in highly unstable and heterogeneous formulations with limited clinical applicability. The aim of this study was to design, formulate, and carefully characterize a stable CD8-inducing adjuvant based on the TLR3 ligand polyinosinic-polycytidylic acid [poly(I:C)] incorporated into a cationic adjuvant system (CAF01) composed of dimethyldioctadecylammonium (DDA) and trehalose 6,6'-dibehenate (TDB). For this purpose, a modified double emulsion solvent evaporation method was investigated for complexation of high amounts of anionic poly(I:C) to gel-state DDA/TDB liposomes. Addition of a volatile, water-miscible co-solvent (ethanol) to the outer water phase enabled preparation of colloidally stable liposomes, presumably by reducing the poly(I:C)-enhanced rigidity of the lipid bilayer. Cryo-transmission electron microscopy (TEM) revealed the formation of unilamellar as well as multilamellar liposomes, the latter suggesting that poly(I:C) is intercalated between the membrane bilayers in an onion-like structure. Finally, immunization of mice with the model antigen ovalbumin (OVA) and DDA/TDB/poly(I:C) liposomes induced a remarkably strong, antigen-specific CD8+ T-cell response, which was maintained for more than two months. Importantly, whereas injection of soluble poly(I:C) led to rapid production of the pro-inflammatory cytokines tumor necrosis factor (TNF)-α and interleukin (IL)-6 in serum, administration of poly(I:C) in complex with the cationic DDA/TDB liposomes prevented this non-specific systemic pro-inflammatory response. These data emphasize the importance of improving the quality of the vaccine formulation to indeed overcome some of the major obstacles for using CD8-inducing agents such as poly(I:C) in future subunit vaccines.
Topics: Adjuvants, Immunologic; Animals; CD8-Positive T-Lymphocytes; Female; Glycolipids; Interferon Inducers; Liposomes; Mice; Mice, Inbred C57BL; Poly I-C; Quaternary Ammonium Compounds
PubMed: 21111765
DOI: 10.1016/j.jconrel.2010.11.021 -
Innate Immunity Jul 2014Many modern vaccines use defined adjuvants to stimulate the innate immune system and shape the adaptive immune response. The exact nature of these innate signals and...
Many modern vaccines use defined adjuvants to stimulate the innate immune system and shape the adaptive immune response. The exact nature of these innate signals and whether immune differentiation can originate within the periphery is not known. Here we used an ovine lymphatic cannulation model to characterise the cellular and transcriptomic profile of the afferent lymph following injection of a liposomal vaccine formulation incorporating diphtheria toxoid and the innate stimulator poly(I:C) over a 78-h period. The response to this vaccine featured an early activation of broad pro-inflammatory pathways (e.g. TLR signalling and inflammasome pathways) and the transient recruitment of granulocytes into the lymph. At 24 h a more monocytic cellular profile arose coinciding with a transition to a specific antiviral response characterised by the up-regulation of genes associated with the receptors typical for the viral mimic, poly(I:C) (e.g. TLR3, RIG-I and MDA5). At the latest time points the up-regulation of IL-17A and IL-17F suggested that Th17 cells may participate in the earliest adaptive response to this vaccine. These data provide the most comprehensive picture of the cellular and molecular mechanisms that link the periphery to the draining lymph node following vaccination, and indicate that the immune response is capable of specialising within the periphery.
Topics: Animals; Antiviral Agents; Diphtheria Toxoid; Granulocytes; Immunity, Innate; Inflammasomes; Interleukin-17; Liposomes; Lymph; Poly I-C; Sheep; Th17 Cells; Toll-Like Receptors; Up-Regulation; Vaccination
PubMed: 24045338
DOI: 10.1177/1753425913501213 -
Experimental Eye Research Apr 1974
Topics: Animals; Drug Therapy, Combination; Female; Idoxuridine; Keratitis, Dendritic; Keratoconjunctivitis; Male; Ophthalmic Solutions; Poly I-C; Rabbits
PubMed: 4834043
DOI: 10.1016/0014-4835(74)90114-6 -
The Journal of Biological Chemistry Apr 1990The technique of photoaffinity labeling has been applied to the double-stranded RNA (dsRNA)-dependent enzyme 2',5'-oligoadenylate (2-5A) synthetase to provide a means...
8-Azido double-stranded RNA photoaffinity probes. Enzymatic synthesis, characterization, and biological properties of poly(I,8-azidoI).poly(C) and poly(I,8-azidoI).poly(C12U) with 2',5'-oligoadenylate synthetase and protein kinase.
The technique of photoaffinity labeling has been applied to the double-stranded RNA (dsRNA)-dependent enzyme 2',5'-oligoadenylate (2-5A) synthetase to provide a means for the examination of RNA-protein interaction(s) in the dsRNA allosteric binding domain of this enzyme. The synthesis, characterization, and biological properties of the photoaffinity probe poly[( 32P]I,8-azidoI).poly(C) and its mismatched analog poly[( 32P]I,8-azidoI).poly(C12U), which mimic the parent molecules poly(I).poly(C) and poly(I).poly(C12U), are described. The efficacy of poly[( 32P]I,8-azidoI).poly(C) and poly[( 32P]I,8-azidoI).poly(C12U) as allosteric site-directed activators is demonstrated using highly purified 2-5A synthetase from rabbit reticulocyte lysates and from extracts of interferon-treated HeLa cells. The dsRNA photoprobes activate these two 2-5A synthetases. Saturation of 2-5A synthetase is observed at 6 x 10(-4) g/ml poly[( 32P]I,8-azidoI).poly(C) following photolysis for 20 s at 0 degrees C. The photoincorporation of poly[( 32P]I,8-azidoI).poly(C) is specific, as demonstrated by the prevention of photoincorporation by native poly(I).poly(C). DNA, poly(I), and poly(C) are not competitors of poly[( 32P]I,8-azidoI).poly(C). Following UV irradiation of 2-5A synthetase with poly[( 32P]I,8-azidoI).poly(C), the reaction mixture is treated with micrococcal nuclease to hydrolyze azido dsRNA that is not cross-linked to the enzyme. A radioactive band of 110 kDa (the same as that reported for native rabbit reticulocyte lysate 2-5A synthetase) is observed following sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography. The specific photolabeling of the 2-5A synthetase suggests that the azido dsRNA is intrinsic to the allosteric binding domain. The utility of poly[( 32P]I,8-azidoI).poly(C) for the detection of dsRNA-dependent binding proteins and the isolation of peptides at or near the allosteric binding site is discussed.
Topics: 2',5'-Oligoadenylate Synthetase; Affinity Labels; Allosteric Site; Animals; Azides; Binding, Competitive; Enzyme Activation; HeLa Cells; Humans; Micrococcal Nuclease; Photochemistry; Poly I-C; Poly U; Protein Kinases; RNA, Double-Stranded; Rabbits; Reticulocytes; Ultraviolet Rays
PubMed: 2318823
DOI: No ID Found