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Stem Cells (Dayton, Ohio) Feb 2014Mesenchymal stem cells (MSCs) are attractive candidates for clinical therapeutic applications. Recent studies indicate MSCs express active Toll-like receptors (TLRs),...
Mesenchymal stem cells (MSCs) are attractive candidates for clinical therapeutic applications. Recent studies indicate MSCs express active Toll-like receptors (TLRs), but their effect on MSCs and the underlying mechanisms remain unclear. In this study, we found that, after treating human umbilical cord MSCs with various TLR ligands, only TLR3 ligand, poly(I:C), could significantly increase the expression of cyclooxygenase-2 (COX-2). Furthermore, poly(I:C) could enhance MSCs' anti-inflammatory effect on macrophages. Next, we focused on the regulatory roles of microRNAs (miRNAs) in the process of poly(I:C) activating MSCs. Our experiments indicated that miR-143 expression was significantly decreased in MSCs with poly(I:C) treatment, and the expression level of miR-143 could regulate the effect of poly(I:C) on MSCs' immunosuppressive function. Subsequent results showed that the reporter genes with putative miR-143 binding sites from the transforming growth factor-β-activated kinase-1 (TAK1) and COX-2 3' untranslated regions were downregulated in the presence of miR-143. In addition, mRNA and protein expression of TAK1 and COX-2 in MSCs was also downregulated with miR-143 overexpression, suggesting that TAK1 and COX-2 are target genes of miR-143 in MSCs. Consistent with miR-143 overexpression, TAK1 interference also attenuated MSCs' immunosuppressive function enhanced by poly(I:C). Additionally, it was shown that TLR3-activated MSCs could improve survival in cecal ligation and puncture (CLP)-induced sepsis, while miR-143 overexpression reduced the effectiveness of this therapy. These results proved that poly(I:C) improved the immunosuppressive abilities of MSCs, revealed the regulatory role of miRNAs in the process, and may provide an opportunity for potential novel therapies for sepsis.
Topics: Cell Differentiation; Cyclooxygenase 2; Humans; Immunosuppression Therapy; Macrophages; Mesenchymal Stem Cell Transplantation; Mesenchymal Stem Cells; MicroRNAs; Poly I-C; RNA, Messenger; Sepsis; Toll-Like Receptor 3; Umbilical Cord
PubMed: 24105952
DOI: 10.1002/stem.1543 -
Proceedings of the Society For... Feb 1975The present report confirms the findings by Steinberg et al. (9) that repeated intraperitoneal injections of poly I:C (3 mug/g, three times per wk, 40-52 doses) enhanced...
The present report confirms the findings by Steinberg et al. (9) that repeated intraperitoneal injections of poly I:C (3 mug/g, three times per wk, 40-52 doses) enhanced the incidence and severity of glomerular lesions that occur spontaneously in NZG/NZW mice and also increased the development of circulating antibody against nucleic acids. This effect was minimal when only six intraperitoneal doses were given in 1 mug/g amount at weekly intervals. Intranasal administration of poly I:C (0.2 mug/g, three times per wk, 40 doses) or six doses of the drug (1 mug/g weekly) caused no apparent potentiation of glomerular response. ICR/Ha mice, which do not suffer from the spontaneously occurring disease, were uneffected by poly I:C treatment except for occasional development of antibody against poly I:C or DNA.
Topics: Administration, Intranasal; Animals; Antibodies; DNA; Drug Evaluation, Preclinical; Female; Injections, Intraperitoneal; Kidney Glomerulus; Male; Mice; Mice, Inbred ICR; Mice, Inbred NZB; Poly I-C; RNA
PubMed: 1121498
DOI: 10.3181/00379727-148-38565 -
Research Communications in Chemical... Sep 1984Thiolation of position 5 of some of the cytosine bases in polycytidylic acid results in the formation of mercaptopolycytidylic acid (MPC). Annealing of MPC to...
Thiolation of position 5 of some of the cytosine bases in polycytidylic acid results in the formation of mercaptopolycytidylic acid (MPC). Annealing of MPC to polyinosinic acid (Poly I) results in the formation of double-stranded Poly I-MPC. In this study we investigated the interferon inducing ability, in vivo toxic effects, effect on DNA synthesis, and the effects in human tumor cell lines of Poly I-MPC. Poly I-MPC was capable of inducing human alpha, beta and gamma interferons in the appropriate cell systems. In vivo toxicity was measured in mice, guinea pigs, and rabbits according to FDA guidelines. Weight loss and lethal and pyrogenic effects were markedly lower in Poly I-MPC treated animals than in those that received unmodified Poly I-Poly C. In contrast to the lack of an effect of Poly I-Poly C in human lymphocytes, Poly I-MPC inhibited DNA synthesis. It also inhibited colony formation and was cytotoxic in several human tumor cell lines. Poly I-MPC's ability to induce human alpha, beta and gamma interferons, to inhibit DNA synthesis and its effects in human tumor cell lines demonstrate the potential of this drug for future clinical studies, both as an antiviral and antitumor agent.
Topics: Animals; Antineoplastic Agents; Antiviral Agents; Cell Division; Cell Line; Cell Survival; Colony-Forming Units Assay; DNA; Fibroblasts; Guinea Pigs; Humans; Interferon Inducers; Interferon Type I; Interferon-gamma; Lymphocytes; Mice; Neoplasms, Experimental; Poly I-C
PubMed: 6438740
DOI: No ID Found -
Journal of Fish Diseases Mar 2012It was recently reported that Poly(I:C) immunization with live nervous necrosis virus (NNV) confers protection in sevenband grouper, Epinephelus septemfasciatus...
It was recently reported that Poly(I:C) immunization with live nervous necrosis virus (NNV) confers protection in sevenband grouper, Epinephelus septemfasciatus (Thunberg), from NNV infection. In the present study, we conducted field tests with sevenband grouper for the evaluation of Poly(I:C) immunization efficacy. In the first experiment, sevenband grouper were immunized with NNV followed by Poly(I:C) administration 7 weeks before natural occurrence of viral nervous necrosis (VNN). Survival rate of the naïve fish was 71.0%, whereas that of the immunized fish was 99.8%. In the second experiment, sevenband grouper were immunized 10 months before VNN occurrence and survival rate of the non-treated and vaccinated fish was 79.5% and 97.5%, respectively. In the third experiment, we administered Poly(I:C) to sevenband grouper at 20 days after natural occurrence of VNN. The survival rate of the non-treated fish was 9.8%, whereas that of fish administered Poly(I:C) was 93.7%. Based on these results, it was concluded that Poly(I:C) immunization conferred protection in fish against NNV infection in field tests and the protection lasted more than 10 months. Furthermore, even after occurrence of VNN, fish mortality could be reduced by Poly(I:C) administration and there was an unexpected curative effect on VNN-affected fish.
Topics: Animals; Fish Diseases; Fishes; Immunization; Nodaviridae; Poly I-C; RNA Virus Infections; Time Factors; Viral Vaccines
PubMed: 22239254
DOI: 10.1111/j.1365-2761.2011.01334.x -
Analytical Biochemistry Jan 2019Toll-like receptor 3 (TLR3), a pathogen recognition receptor of the innate immune response, recognizes and is activated by double-stranded RNA (dsRNA), which is...
Toll-like receptor 3 (TLR3), a pathogen recognition receptor of the innate immune response, recognizes and is activated by double-stranded RNA (dsRNA), which is indicative of viral exposure. A sensor design exercise was conducted, using surface plasmon resonance detection, through the examination of several immobilization approaches for TLR3 as a biorecognition element (BRE) onto a modified gold surface. To examine the TLR3-dsRNA interaction a synthetic analogue mimic, poly (I:C), was used. The interaction binding characteristics were determined and compared to literature data to establish the optimal immobilization method for the TLR3 BRE. A preliminary evaluation of the efficacy of the selected TLR3 surface as a broad-spectrum viral biosensor was also performed. Amine-coupling was found to be the most reliable method for manufacturing repeatable and consistent TLR3 BRE sensor surfaces, although this immobilization schema is not tailored to place the receptor in a spatially-specific orientation. The equilibrium dissociation constant (K) measured for this immobilized TLR3-poly (I:C) interaction was 117 ± 3.30 pM. This evaluation included a cross-reactivity study using a selection of purified E. coli and synthetic double- and single-stranded nucleic acids. The results of this design exercise and ligand binding study will inform future work towards the development of a broad-spectrum viral sensor device.
Topics: Biosensing Techniques; Nucleic Acids; Poly I-C; Protein Binding; Surface Plasmon Resonance; Toll-Like Receptor 3
PubMed: 29842862
DOI: 10.1016/j.ab.2018.05.023 -
Vaccine Jan 2009We have recently reported that the toll-like receptor 3 (TLR3) agonist poly(I:C) induces adjuvant effects to post vaccination CD8+ T cells responses through rapid...
We have recently reported that the toll-like receptor 3 (TLR3) agonist poly(I:C) induces adjuvant effects to post vaccination CD8+ T cells responses through rapid induction of innate mediators, including NK cells, macrophages, dendritic cells (DCs), and inflammatory cytokines. However, whether this TLR3 agonist directly targets CD8+ T cells needs to be carefully investigated. In this study, we found that optimal post vaccination CD8+ T cell responses to ex vivo DC-based vaccination requires triggering of TLR3 signaling pathway in DCs in vitro as well as in the recipient host, indicating a role for other cell types. Real-time PCR analysis revealed that TLRs (TLR1-TLR13) are expressed in purified (>99% pure) CD4+ and CD8+ T cells from C57BL/6 and BALB/c mice, where the magnitude of the expression was strain and cell type dependent. In vitro, treatment of these purified T cells with poly(I:C) modulated the expression of TLRs including TLR3. Furthermore, non-specific and antigen-specific stimulation of CD8+ T cells by phorbol myristate acetate and MHC class I peptide-pulsed splenocytes, respectively, modulated TLR expression in purified CD4+ and CD8+ T cells. Importantly, brief conditioning of purified naïve TCR transgenic OT-1 (CD8+) T cells in vitro with poly(I:C) induced activation of these cells in absence of antigen stimulation. Interestingly, when these in vitro poly(I:C)-conditioned OT-1 cells were adoptively transferred into naïve recipient followed by peptide vaccination, they showed superior expansion and activation to their naïve counterparts. These results suggest that CD8+ T cells can be activated by triggering their TLR3. Furthermore, the data support the notion of direct involvement of TLRs in adaptive immune responses.
Topics: Adoptive Transfer; Animals; CD8-Positive T-Lymphocytes; Dendritic Cells; Lymphocyte Activation; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Transgenic; Poly I-C; Toll-Like Receptor 3; Vaccination
PubMed: 19027047
DOI: 10.1016/j.vaccine.2008.11.013 -
BMC Genomics Aug 2020Manila clam (Ruditapes philippinarum) is a worldwide commercially important marine bivalve species. In recent years, however, microbial diseases caused high economic...
BACKGROUND
Manila clam (Ruditapes philippinarum) is a worldwide commercially important marine bivalve species. In recent years, however, microbial diseases caused high economic losses and have received increasing attention. To understand the molecular basis of the immune response to pathogen-associated molecular patterns (PAMPs) in R. philippinarum, transcriptome libraries of clam hepatopancreas were constructed at 24 h post-injection with Lipopolysaccharide (LPS), peptidoglycan (PGN), and polyinosinic-polycytidylic acid (poly(I:C)) and phosphate-buffered saline (PBS) control by using RNA sequencing technology (RNA-seq).
RESULTS
A total of 832, 839, and 188 differentially expressed genes (DEGs) were found in LPS, PGN, and poly(I:C) challenge group compared with PBS control, respectively. Several immune-related genes and pathways were activated in response to the different PAMPs, suggesting these genes and pathways might specifically participate in the immune response to pathogens. Besides, the analyses provided useful complementary data to compare different PAMPs challenges in vivo. Functional enrichment analysis of DEGs demonstrated that PAMPs responsive signal pathways were related to apoptosis, signal transduction, immune system, and signaling molecules and interaction. Several shared or specific DEGs response to different PAMPs were revealed in R. philippinarum, including pattern recognition receptors (PRRs), antimicrobial peptides (AMPs), interferon-induced proteins (IFI), and some other immune-related genes were found in the present work.
CONCLUSIONS
This is the first study employing high throughput transcriptomic sequencing to provide valuable genomic resources and investigate Manila clam response to different PAMPs through in vivo challenges with LPS, PGN, and poly(I:C). The results obtained here provide new insights to understanding the immune characteristics of R. philippinarum response to different PAMPs. This information is critical to elucidate the molecular basis of R. philippinarum response to different pathogens invasion, which potentially can be used to develop effective control strategies for different pathogens.
Topics: Animals; Bivalvia; Lipopolysaccharides; Pathogen-Associated Molecular Pattern Molecules; Peptidoglycan; Poly I-C; RNA-Seq; Sequence Analysis, RNA
PubMed: 32738896
DOI: 10.1186/s12864-020-06914-2 -
Fish & Shellfish Immunology Dec 2017Crustacean hepatopancreas regulates metabolic processes, biogenesis and innate immune processes, and the knowledge on its immune genes are crucial to understand...
Crustacean hepatopancreas regulates metabolic processes, biogenesis and innate immune processes, and the knowledge on its immune genes are crucial to understand antimicrobial mechanisms. In this study, we reported the transcriptomic profile of Procambarus clarkii hepatopancreas after poly I:C administration using high-throughput sequencing. Following de novo assembly 56,716 unigene sequences with an average length of 810 bp was obtained. The unigene sequences were annotated to three ontologies including cellular components, biological processes and molecular functions, further 56,716 unigene sequences were mapped to 25 COG categories. A total of 2497 differentially expressed genes (DEGs) were identified following the comparative analysis between poly I:C treated and control group, and then KEGG enrichment analysis were performed to detect immune related pathways. Quantitative real time polymerase chain reaction showed that the selected DEGs significantly up-regulated following poly I:C administration in comparison to control group. The transcriptomic sequence information will improve the knowledge of this economically important crustacean, and will shed light on its antiviral immune mechanisms.
Topics: Animals; Arthropod Proteins; Astacoidea; Gene Expression Profiling; Hepatopancreas; Poly I-C; Transcriptome
PubMed: 29017948
DOI: 10.1016/j.fsi.2017.10.010 -
Journal of Biological Response Modifiers Dec 1985Polyinosinic-polycytidylic acid stabilized with poly-L-lysine in carboxymethylcellulose [poly(I,C)-LC] significantly augmented natural killer (NK) cell activity in...
Polyinosinic-polycytidylic acid stabilized with poly-L-lysine in carboxymethylcellulose [poly(I,C)-LC] significantly augmented natural killer (NK) cell activity in several tissues. Macrophage (M phi) tumoricidal activity was also markedly increased. Both effector cells were active for 9 days. Poly(I,C)-LC also increased effector cell response in vitro. Injections of poly(I,C)-LC resulted in elevated effector cell responses in four of five routes tested. Treatment with poly(I,C)-LC led to an earlier reconstitution of bone marrow cells, NK cell activity, and M phi effector cell activity in mice pretreated with cyclophosphamide (Cytoxan). Combined treatment of MBL-2 tumor cells with cytoreductive chemotherapy and poly(I,C)-LC resulted in an enhanced therapeutic response.
Topics: Animals; Antineoplastic Agents; Carboxymethylcellulose Sodium; Cyclophosphamide; Drug Therapy, Combination; Female; In Vitro Techniques; Killer Cells, Natural; Kinetics; Leukemia, Experimental; Macrophages; Methylcellulose; Mice; Poly I-C; Polylysine
PubMed: 4087033
DOI: No ID Found -
Fish & Shellfish Immunology Aug 2015Gene expression profiling of poly (I:C)-treated Japanese flounder, Paralichthys olivaceus, under different temperatures was investigated using microarray analysis. The...
Gene expression profiling of poly (I:C)-treated Japanese flounder, Paralichthys olivaceus, under different temperatures was investigated using microarray analysis. The response was analyzed in spleen tissue at 3 and 24 h post injection (hpi) at 15 °C and 25 °C. A large number of genes in fish treated with poly (I:C) at 25 °C were expressed at 3 hpi, whereas the expression profiles at 24 hpi appeared to be similar to those of the controls. Cluster analysis of the different expression profiles showed three distinct groups of up-regulated genes in fish reared at 15 °C. These were early (3 hpi), early-to-late (3 and 24 hpi), and late (24 hpi) up-regulated genes. These genes included type I IFN-related genes and inflammatory genes. Among the up-regulated genes, most of the type I IFN-related genes played early-to-late- and late-responding genes at 15 °C but early-responding genes at 25 °C. Thus, several up-regulated genes in these groups from the microarray result were further verified by qPCR. These results indicate that the type I IFN gene expressions of P. olivaceus treated with poly (I:C) can be regulated in a temperature-dependent manner.
Topics: Animals; Cluster Analysis; Flatfishes; Gene Expression Regulation; Oligonucleotide Array Sequence Analysis; Poly I-C; Polymerase Chain Reaction; Temperature
PubMed: 26052011
DOI: 10.1016/j.fsi.2015.05.036