Comparison of the efficacy of Poly(I:C) immunization with live vaccine and formalin-killed vaccine against viral hemorrhagic septicemia virus (VHSV) in olive flounder (Paralichthys olivaceus)

Kim, H.J.,Park, J.S.,Choi, M.C.,Kwon, S.R. Academic Press 2016 Fish & Shellfish Immunology Vol.48 No.-

<P>Viral hemorrhagic septicemia (VHS) in olive flounder, Paralichthys olivaceus, causes significant economic loss for the flounder aquaculture industry in Korea. In this study, the immunogenicity of Poly(I:C) immunization with a live vaccine against the VHS virus (VHSV) was compared with that of a formalin-treated vaccine in the olive flounder. In vaccine trial I, fish pre-injected with Poly(I:C) were highly protected from VHSV infection 2 d later (survival rate: 96%) and the surviving fish (Poly(I:C)-VHSV group) showed a 100% survival rate against VHSV re-challenge. Mortality in fish pre-injected with diethylpyrocarbonate-treated water followed by injection with formalin-treated VHSV was only 2% (1 of 50 fish), whereas survivors (DEPC-FT VHSV group) showed an 80% survival rate. In vaccine trial II, 100% survival was observed in all Poly(I:C) vaccination groups Poly(1:C)-VHSV 6, Poly(1:C)-VHSV 5, and Poly(I:C)-VHSV 4. In contrast, the survival rates of the groups administered the formalin-treated VHSV at a dose of 10(6), 10(5), and 10(4) TCID50 100 mu L-1 fish(-1) (DEPC-FT VHSV 6, DEPC-FT VHSV 5, and DEPC-FT VHSV 4) were only 8%, 12%, and 12%, respectively. The differences in the survival rates of the formalin-treated vaccine groups in trial I and trial II were attributed to the difference in the formalin-treatment period: the formalin-treated VHSV administered in trial I was not completely inactivated and worked as a live vaccine, which explains the 80% survival rate against VHSV challenge. Specific antibodies against VHSV were detected in sera from all vaccinated survivors, except the DEPC-VHSV 4 group. Furthermore, the specific antibody titers of fish vaccinated with the live and dead VHSV vaccines were similar, but the protective effects of the live and dead vaccines varied considerably. Our findings show that Poly(I:C) immunization with the live vaccine offers better protection than the formalin-treated vaccine against VHS in olive flounder and revealed that antibody levels are not a reliable indicator of the protective effect of the vaccine against the pathogen. In the future, elements of T cell immunity may be used as a means of evaluating the protective efficacy of a vaccine against VHSV instead of ELISA. (C) 2015 Elsevier Ltd. All rights reserved.</P>

Poly I:C inhibits cell proliferation and enhances the growth inhibitory effect of paclitaxel in oral sqaumous cell carcinoma

Park, Jong-Hwan,Jeon, Do-In,Yoon, Hyo-Eun,Kwon, Seong-Min,Kim, Soo-A,Ahn, Sang-Gun,Yoon, Jung-Hoon Informa Healthcare 2012 Acta odontologica scandinavica Vol.70 No.3

<P><B><I>Objective.</I></B> Toll-like receptors (TLR) signaling has dual effect of promoting tumor progression and anti-cancer property. This study was designed to determine the effect of polyinosinic-polycytidilic acid (poly I:C), a TLR3 agonist, on the proliferation of oral cancer cells. <B><I>Materials and methods:</I></B> Human oral squamous cell carcinoma cell lines, YD-10B and YD-8, were used. TLRs expression was examined by RT-PCR and IL-8 production by poly I:C was examined by ELISA. Cell proliferation was determined by MTT assay. Flow cytometry and Western blot analysis were performed to determine the molecular mechanism of poly I:C-induced cell death. <B><I>Results.</I></B> TLR3 was functionally expressed in YD-10B and YD-8 cells. Treatment of poly I:C inhibited the cell growth in a dose-dependent manner. Flow cytometry and Western blot analysis revealed that poly I:C induced apoptosis via a mitochondria-dependent pathway. In addition, combination treatment with poly I:C and paclitaxel more significantly inhibited cell proliferation compared with poly I:C or paclitaxel alone. <B><I>Conclusions.</I></B> Poly I:C effectively inhibits oral cancer cell proliferation and can be considered as a candidate to improve the inhibitory effect of anti-cancer drugs.</P>

Innate immune responses against rock bream iridovirus (RBIV) infection in rock bream (<i>Oplegnathus fasciatus</i>) following poly (I:C) administration

Jung, Myung-Hwa,Jung, Sung-Ju ACADEMIC PRESS LTD 2017 FISH AND SHELLFISH IMMUNOLOGY Vol.71 No.-

<P><B>Abstract</B></P> <P>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<SUP>4</SUP> and 8.95 × 10<SUP>5</SUP>/μl, respectively), 7 (3.67 × 10<SUP>5</SUP> and 8.81 × 10<SUP>6</SUP>/μl, respectively) and 10 days post infection (dpi) (1.26 × 10<SUP>5</SUP> and 6.02 × 10<SUP>7</SUP>/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.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Poly (I:C) inhibited virus replication in spleen, kidney, liver and blood of rock bream. </LI> <LI> TLR3 and IL1β were highly expressed in the poly (I:C)+virus-injected group. </LI> <LI> Mx and ISG15 were significantly expressed earlier (from 2 dpi) in the poly (I:C)+virus-injected group than in the virus-injected group (from 4 dpi). </LI> </UL> </P>

Molecular cloning and characterization of Toll-like receptor 3 in Japanese flounder, <i>Paralichthys olivaceus</i>

Hwang, Seong Don,Ohtani, Maki,Hikima, Jun-ichi,Jung, Tae Sung,Kondo, Hidehiro,Hirono, Ikuo,Aoki, Takashi Elsevier 2012 DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY Vol.37 No.1

<P><B>Highlights</B></P><P>► We cloned the Japanese flounder TLR3 (JfTLR3) gene. ► Gene expression of JfTLR3 was up-regulated in PBLs after stimulation with poly I:C and CpG ODN 1668. ► JfTLR3 gene was induced in different tissues following infection with VHSV. ► Intracellular poly I:C in JfTLR3-overexpressing YO-K cells activates IFN and NF-κB pathways. ► In JfTLR3-overexpressing HINAE cells, extracellular poly I:C activates IFN and NF-κB pathways.</P> <P><B>Abstract</B></P><P>Mammalian Toll-like receptor 3 (TLR3) recognizes extracellular and intracellular viral dsRNA, and then initiates signaling cascades leading to NF-κB activation and interferon (IFN) production. To understand the roles of TLR3 in the fish immune system, TLR3 gene (JfTLR3) was identified from Japanese flounder (<I>Paralichthys olivaceus</I>), which consisted of 4 exons and 3 introns. Its expression in peripheral blood leukocytes increased upon stimulation with poly I:C and CpG ODN 1668. Exposure to viral hemorrhagic septicemia virus increased expression of JfTLR3 in the blood, liver, head kidney and spleen. Intracellular poly I:C stimulation in JfTLR3-overexpressing YO-K cells significantly induced IFN-inducible and NF-κB-regulated genes. NF-κB activity in JfTLR3-overexpressing YO-K cells was significantly induced by intracellular poly I:C while expression of IFN-inducible genes and NF-κB reporter activity in JfTLR3-overexpressing HINAE cells increased upon stimulation by extracellular poly I:C. These results suggest that JfTLR3 plays an important role in the induction of antiviral immune response.</P>

Inhibition of Poly(I:C)-Induced Inflammation by Salvianolic Acid A in Skin Keratinocytes

( Qing-ling Zhang ),( Ri-hua Jiang ),( Xue Mei Li ),( Jung-woo Ko ),( Chang Deok Kim ),( Ming Ji Zhu ),( Jeung-hoon Lee ) 대한피부과학회 2019 Annals of Dermatology Vol.31 No.3

Background: Skin keratinocytes participate actively in inducing immune responses when external pathogens are introduced, thereby contributing to elimination of pathogens. However, in condition where the excessive inflammation is occurred, chronic skin disease such as psoriasis can be provoked. Objective: We tried to screen the putative therapeutics for inflammatory skin disease, and found that salvianolic acid A (SAA) has an inhibitory effect on keratinocyte inflammatory reaction. The aim of this study is to demonstrate the effects of SAA in poly(I:C)-induced inflammatory reaction in skin keratinocytes. Methods: We pre-treated keratinocytes with SAA then stimulated with poly(I:C). Inflammatory reaction of keratinocytes was verified using real-time polymerase chain reaction, enzyme-linked immunosorbent assay and Western blot. Results: When skin keratinocytes were pre-treated with SAA, it significantly inhibited poly (I:C)-induced expression of inflammatory cytokines including interleukin (IL)-1β, IL-6, IL-8, tumor necrosis factor-α, and CCL20. SAA inhibited poly(I:C)-induced activation of nuclear factor-κB signaling. And SAA also inhibited inflammasome activation, evidenced by decrease of IL-1β secretion. Finally, SAA markedly inhibited poly(I:C)-induced NLRP3 expression. Conclusion: These results demonstrate that SAA has an inhibitory effect on poly(I:C)-induced inflammatory reaction of keratinocytes, suggesting that SAA can be developed for the treatment of inflammatory skin diseases such as psoriasis. (Ann Dermatol 31(3) 279∼285, 2019)

Induction of alopecia areata in C3H/HeJ mice using polyinosinic-polycytidylic acid (poly[I:C]) and interferon-gamma

( Jung-min Shin ),( Soo Jung Kim ),( Xue Mei Li ),( Jin Gwi Yoo ),( Dong-kyun Hong ),( Jin-hyup Lee ),( Chong Won Choi ),( Kyung-duck Park ),( Chang Deok Kim ),( Young-joon Seo ),( Jeung-hoon Lee ),( 대한피부과학회 2018 대한피부과학회 학술발표대회집 Vol.70 No.2

Background: Alopecia areata (AA) is a chronic, relapsing hair-loss disorder that is considered to be a T-cell-mediated autoimmune disease. Several animal models for AA have been created to investigate the pathophysiology and screen for effective therapeutic targets. Objectives: As C3H/HeJ mice develop AA spontaneously in a low frequency, a novel animal model is needed to establish an AA-like condition faster and more conveniently. In this study, we present a novel non-invasive AA rodent model that avoids skin or lymph-node cell transfer. Methods: We simply injected C3H/HeJ mice subcutaneously with interferon-gamma (IFNγ) along with polyinosinic:polycytidylic acid (poly[I:C]), a synthetic dsRNA, to initiate innate immunity via inflammasome activation. Results: Approximately 80% of the IFNγ and poly(I:C) co-injected mice showed patchy AA lesions after 8 weeks. None of the mice displayed hair loss in the IFNγ or poly(I:C) solely injection group. Immunohistochemical staining of the AA lesions revealed increased infiltration of CD4+ and CD8+ cells infiltration around the hair follicles. IFNγ and poly(I:C) increased the expression of NLRP3, IL-1b, CXCL9, CXCL10, and CXCL11 in mouse skin. Conclusion: Taken together, these findings indicate a shorter and more convenient means of AA animal model induction and demonstrate that inflammasome-activated innate immunity is important in AA pathogenesis.

Inhibition of poly(I:C)-induced inflammation by salvianolic acid A in skin keratinocytes

( Su-hyuk Yim ),( Qing-ling Zhang ),( Xue Mei Li ),( Jin Gwi Yoo ),( Dong-kyun Hong ),( Jin-hyup Lee ),( Chong Won Choi ),( Kyung Duck Park ),( Young Lee ),( Chang Deok Kim ),( Young-joon Seo ),( Jeun 대한피부과학회 2018 대한피부과학회 학술발표대회집 Vol.70 No.2

Background: Skin keratinocytes participate actively in inducing immune responses when external pathogens are introduced, thereby contributing to elimination of pathogens. However, in condition where the excessive inflammation is occurred, chronic skin disease such as psoriasis can be provoked. Objectives: We tried to screen the putative therapeutics for inflammatory skin disease, and found that salvianolic acid A (SAA) has an inhibitory effects on keratinocyte inflammatory reaction. The aim of this study is to demonstrate the effects of SAA in poly(I:C)-induced inflammatory reaction in skin keratinocytes. Methods: The keratinocytes were pretreated with SAA then stimulated with poly(I:C). Inflammatory reaction of keratinocytes, then we verified using RT-PCR, ELISA and Western blot. Results: When skin keratinocytes were pre-treated with SAA, it significantly inhibited poly(I:C)-induced expression of inflammatory cytokines including IL-1β, IL-6, IL-8, TNF-α, and CCL20. SAA inhibited poly(I:C)-induced activation of NF-κB signaling. And SAA also inhibited inflammasome activation, evidenced by decrease of IL-1β secretion. Finally, SAA markedly inhibited poly(I:C)-induced NLRP3 expression. Conclusion: These results demonstrate that SAA has an inhibitory effect on poly(I:C)-induced inflammatory reaction of keratinocytes, suggesting that SAA can be developed for the treatment of inflammatory skin diseases such as psoriasis.

Anti-inflammatory mechanism of lonchocarpine in LPS- or poly(I:C)-induced neuroinflammation

Jeong, Yeon-Hui,Park, Jin-Sun,Kim, Dong-Hyun,Kang, Jihee Lee,Kim, Hee-Sun Elsevier 2017 PHARMACOLOGICAL RESEARCH Vol.119 No.-

<P><B>Abstract</B></P> <P>Neuroinflammation plays an important role in the progression of various neurodegenerative diseases. In this study, we investigated the anti-inflammatory effects of lonchocarpine, a natural compound isolated from <I>Abrus precatorius</I>, under <I>in vitro</I> and <I>in vivo</I> neuroinflammatory conditions induced by challenge with lipopolysaccharide (LPS)- or polyinosinic-polycytidylic acid (poly(I:C)). Lonchocarpine suppressed the expression of iNOS and proinflammatory cytokines in LPS or poly(I:C)-stimulated BV2 microglial cells. These anti-inflammatory effects were verified in brains of mice with systemic inflammation induced by administration of LPS or poly(I:C). Lonchocarpine reduced the number of Iba-1-positive activated microglia, and suppressed the mRNA expression of various proinflammatory markers in the cortex of LPS- or poly(I:C)-injected mice. Molecular mechanistic experiments showed that lonchocarpine inhibited NF-κB activity by reducing the phosphorylation and degradation of IκBα in LPS- or poly(I:C)-stimulated BV2 cells. Analysis of further upstream signaling pathways in LPS-stimulated microglia showed that lonchocarpine inhibited the phosphorylation of IκB kinase and TGFβ-activated kinase 1 (TAK1). Moreover, lonchocarpine suppressed the interaction of myeloid differentiation factor 88 (MyD88) and intereleukin-1 receptor-associated kinase 4 (IRAK4). These data suggest that toll-like receptor 4 downstream signals such as MyD88/IRAK4-TAK1-NF-κB are at least partly involved in the anti-inflammatory mechanism of lonchocarpine in LPS-stimulated microglia. Its strong anti-inflammatory effects may make lonchocarpine an effective preventative drug for neuroinflammatory disorders that are associated with systemic inflammation.</P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

Celastrol suppresses expression of adhesion molecules and chemokines by inhibiting JNK-STAT1/NF-кB activation in poly(I:C)-stimulated astrocytes

( Soo Yeon An ),( Gi Soo Youn ),( Hyejin Kim ),( Soo Young Choi ),( Jinseu Park ) 생화학분자생물학회 2017 BMB Reports Vol.50 No.1

In the central nervous system, viral infection can induce inflammation by up-regulating pro-inflammatory mediators that contribute to enhanced infiltration of immune cells into the central nervous areas. Celastrol is known to exert various regulatory functions, including anti-microbial activities. In this study, we investigated the regulatory effects and the mechanisms of action of celastrol against astrocytes activated with polyinosinic-polycytidylic acid (poly(I:C)), a synthetic dsRNA, as a model of pro-inflammatory mediated responses. Celastrol significantly inhibited poly(I:C)-induced expression of adhesion molecules, such as ICAM-1/VCAM-1, and chemokines, such as CCL2, CXCL8, and CXCL10, in CRT-MG human astroglioma cells. In addition, celastrol significantly suppressed poly(I:C)- induced activation of JNK MAPK and STAT1 signaling pathways. Furthermore, celastrol significantly suppressed poly(I:C)- induced activation of the NF-кB signaling pathway. These results suggest that celastrol may exert its regulatory activity by inhibiting poly(I:C)-induced expression of pro-inflammatory mediators by suppressing activation of JNK MAPK-STAT1/ NF-кB in astrocytes. [BMB Reports 2017; 50(1): 25-30]

넙치(Paralichthys olivaceus) 신장에서 Poly (I:C)의 단기 및 장기적인 면역 효과

성민재,박영진 한국어병학회 2024 한국어병학회지 Vol.37 No.1

Viral diseases cause enormous economic losses to the olive flounder (Paralichthys olivaceus) aqua-culture industry in Korea. This study aimed to identify immune-related genes expressed in the kidney of olive flounder injected with Polyinosinic-polycytidylic acid (Poly (I:C)). Thirty fish were divided into two groups by intraperitoneal injection of 100μl of diethylpyrocarbonate-treated water or poly I:C per fish. Kidney tissues at day 3 and 30 after the injection were used for RNA-seq analysis to identify differentially expressed genes (DEGs). Poly I:C group upregulated il8, cfh, tnfaip2b, c3b.2, ly6d and cd38 genes at 3 days post-injection. Additionally, cd22, ccl34a.3, c9, cxcl19, ccl27a, ccl7, and cfh genes were upregulated at 30 days post-injection. Differential expression gene analysis showed that poly I:C has both short and long-term immune effects in olive flounder. This study provides a theoretical basis for understanding the molecular mechanism of the short and long-term immune effects of poly I:C.