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Tae Sung Jung - One of the best experts on this subject based on the ideXlab platform.

  • Immunostimulatory effect of DDX41 of olive flounder (Paralichthys olivaceus)
    Food and Agricultural Immunology, 2017
    Co-Authors: Jassy Mary S. Lazarte, Young-rim Kim, Jung Seok Lee, Si Won Kim, Jae Wook Jung, Jaesung Kim, Jeong-ho Lee, Tae Sung Jung
    Abstract:

    ABSTRACTThe cellular DEAD-box helicase DDX41, which functions as an initial sensor for cytoplasmic DNA, is involved in the activation of type I interferon (IFN-1) immune response in olive flounder. A plasmid encoding DDX41 (pEF-D) was introduced into flounder cells in vitro and in vivo. Immune responses induced by DDX41 were evaluated by relative quantification value (ΔΔCt) method of RT-qPCR using specific IFN-related gene primers. Results in in vitro, transcript levels of IFN-1, IRF-3, ISG-15 and IL-1β were significantly higher in pEF-D- than pEF-A-transfected cells, with 15-, 4-, 10- and 32-fold changes, respectively. In vivo, elevated expression of these genes was observed in the kidney of pEF-D group on days 1 and 3 post-treatment. The viral challenge test revealed higher survival rate in fish treated with pEF-D (67.5%) than controls, PBS- and pEF-A-treated fish (45%). Conclusively DDX41 elicits a robust IFN-mediated immune response, validating its adjuvant property.

  • enhancement of glycoprotein based dna vaccine for viral hemorrhagic septicemia virus vhsv via addition of the molecular adjuvant DDX41
    Fish & Shellfish Immunology, 2017
    Co-Authors: Jassy Mary S. Lazarte, Young-rim Kim, Jung Seok Lee, Si Won Kim, Jae Wook Jung, Jaesung Kim, Woo Jai Lee, Tae Sung Jung
    Abstract:

    The use of molecular adjuvants to improve the immunogenicity of DNA vaccines has been thoroughly studied in recent years. Glycoprotein (G)-based DNA vaccines had been proven to be effective in combating infection against Rhabdovirus (especially infectious hematopoietic necrosis virus, IHNV) in salmonids. DDX41 is a helicase known to induce antiviral and inflammatory responses by inducing a type I IFN innate immune response. To gain more information regarding G-based DNA vaccines in olive flounder (Paralicthys olivaceus), we tried to develop a more efficient G-based DNA vaccine by adding a molecular adjuvant, DDX41. We designed a DNA vaccine in which the VHSV glycoprotein (G-protein) and DDX41 were driven by the EF-1α and CMV promoters, respectively. Olive flounders were intramuscularly immunized with 1 μg of plasmids encoding the G-based DNA vaccine alone (pEF-G), the molecular adjuvant alone (pEF-D), or the vaccine-adjuvant construct (pEF-GD). At two different time points, 15 and 30 days later, the fish were intraperitoneally infected with VHSV (100 μL; 1 × 106 TCID50/mL). Our assays revealed that the plasmid constructs showed up-regulated expression of IFN-1 and its associated genes at day 3 post-vaccination in both kidney and spleen samples. Specifically, pEF-GD showed statistically higher expression of immune response genes than pEF-G and pEF-D treated group (p < 0.05/p < 0.001). After VHSV challenge, the fish group treated with pEF-GD showed higher survival rate than the pEF-G treated group, though difference was not statistically significant in the 15 dpv challenged group however in the 30 dpv challenged group, the difference was statistically significant (p < 0.05). Together, these results clearly demonstrate that DDX41 is an effective adjuvant for the G-based DNA vaccine in olive flounder. Our novel findings could facilitate the development of more effective DNA vaccines for the aquaculture industry.

  • Enhancement of glycoprotein-based DNA vaccine for viral hemorrhagic septicemia virus (VHSV) via addition of the molecular adjuvant, DDX41.
    Fish & shellfish immunology, 2017
    Co-Authors: Jassy Mary S. Lazarte, Young-rim Kim, Jung Seok Lee, Si Won Kim, Jae Wook Jung, Jaesung Kim, Woo Jai Lee, Tae Sung Jung
    Abstract:

    The use of molecular adjuvants to improve the immunogenicity of DNA vaccines has been thoroughly studied in recent years. Glycoprotein (G)-based DNA vaccines had been proven to be effective in combating infection against Rhabdovirus (especially infectious hematopoietic necrosis virus, IHNV) in salmonids. DDX41 is a helicase known to induce antiviral and inflammatory responses by inducing a type I IFN innate immune response. To gain more information regarding G-based DNA vaccines in olive flounder (Paralicthys olivaceus), we tried to develop a more efficient G-based DNA vaccine by adding a molecular adjuvant, DDX41. We designed a DNA vaccine in which the VHSV glycoprotein (G-protein) and DDX41 were driven by the EF-1α and CMV promoters, respectively. Olive flounders were intramuscularly immunized with 1 μg of plasmids encoding the G-based DNA vaccine alone (pEF-G), the molecular adjuvant alone (pEF-D), or the vaccine-adjuvant construct (pEF-GD). At two different time points, 15 and 30 days later, the fish were intraperitoneally infected with VHSV (100 μL; 1 × 106 TCID50/mL). Our assays revealed that the plasmid constructs showed up-regulated expression of IFN-1 and its associated genes at day 3 post-vaccination in both kidney and spleen samples. Specifically, pEF-GD showed statistically higher expression of immune response genes than pEF-G and pEF-D treated group (p 

  • enhancement of immunogenicity in glycoprotein based dna vaccine by the addition of DDX41 a molecular adjuvant
    Fish & Shellfish Immunology, 2016
    Co-Authors: Jassy Mary S. Lazarte, Young-rim Kim, Jung Seok Lee, Si Won Kim, Jae Wook Jung, Jaesung Kim, Se Pyeong Lim, Tae Sung Jung
    Abstract:

    The emergence of the first commercialized DNA vaccine for fish had prompted for studies that are focused on the development of a more efficacious vaccine that can protect cultured fish against virus infection. This DNA vaccine designed to express viral glycoprotein (G) gene was proven to effectively give protection to several species of fish against rhabdoviruses (ex. viral hemorrhagic septicemia virus, VHSV). Thus, the addition of molecular adjuvant to the DNA vaccine has been considered to be one of the best methods in improving the vaccine’s capability since its presence triggers a more enhanced immune response. DDX41, a cytosolic sensor, has the ability to activate an antiviral cascade of events in response to stimulation of a DNA virus making it a good molecular adjuvant candidate. In the present study, we designed a DNA vaccine consisting of VHSV glycoprotein (VHSVg) and DDX41 which is regulated by EF-1α and CMV promoters, respectively. The aim is to develop a DNA vaccine that can elicit a stronger immune response that could protect the fish against VHSV infection. Four plasmid constructs were prepared for the experiment: pEF-A (empty vector), pEF-G (with VHSVg), pEF-D (with DDX41) and pEF-GD (both VHSVg and DDX41). Expression of individual genes was checked using Western Blot assay, VHSVg in pEF-G and DDX41 in pEF-D were detected using V5 antibody while DDX41 in pEF-GD was detected using myc antibody. Several immune response genes were also evaluated through qPCR to further elucidate the antiviral effect of the plasmid constructs. ∆∆Ct method was utilized to quantify the fold changes of the immune gene transcripts after immunization. qPCR results showed that there are significant increases in the transcript number of IRF-1, ISG-15, and IRF-3 (p< 0.05) as well as IFN-1 (p<0.1) in cells that were immunized with the vaccine-adjuvant construct, pEF-GD, compared to the control plasmid, pEF-A. The results implicated the strength of DDX41 in inducing IFN-mediated immune responses which demonstrates its ability as a molecular adjuvant. This is essential since majority of the DNA vaccines against fish rhabdovirus that we have now are purely based on viral glycoprotein and though it has been proven to be effective, the development of a more efficient DNA vaccine that can improve fish immunity against rhabdovirus infection is significantly beneficial to the aquaculture industry.

  • Enhancement of immunogenicity in glycoprotein-based DNA vaccine by the addition of DDX41, a molecular adjuvant
    Fish & Shellfish Immunology, 2016
    Co-Authors: Jassy Mary S. Lazarte, Young-rim Kim, Jung Seok Lee, Si Won Kim, Jae Wook Jung, Jaesung Kim, Pyeong Lim, Tae Sung Jung
    Abstract:

    The emergence of the first commercialized DNA vaccine for fish had prompted for studies that are focused on the development of a more efficacious vaccine that can protect cultured fish against virus infection. This DNA vaccine designed to express viral glycoprotein (G) gene was proven to effectively give protection to several species of fish against rhabdoviruses (ex. viral hemorrhagic septicemia virus, VHSV). Thus, the addition of molecular adjuvant to the DNA vaccine has been considered to be one of the best methods in improving the vaccine’s capability since its presence triggers a more enhanced immune response. DDX41, a cytosolic sensor, has the ability to activate an antiviral cascade of events in response to stimulation of a DNA virus making it a good molecular adjuvant candidate. In the present study, we designed a DNA vaccine consisting of VHSV glycoprotein (VHSVg) and DDX41 which is regulated by EF-1α and CMV promoters, respectively. The aim is to develop a DNA vaccine that can elicit a stronger immune response that could protect the fish against VHSV infection. Four plasmid constructs were prepared for the experiment: pEF-A (empty vector), pEF-G (with VHSVg), pEF-D (with DDX41) and pEF-GD (both VHSVg and DDX41). Expression of individual genes was checked using Western Blot assay, VHSVg in pEF-G and DDX41 in pEF-D were detected using V5 antibody while DDX41 in pEF-GD was detected using myc antibody. Several immune response genes were also evaluated through qPCR to further elucidate the antiviral effect of the plasmid constructs. ∆∆Ct method was utilized to quantify the fold changes of the immune gene transcripts after immunization. qPCR results showed that there are significant increases in the transcript number of IRF-1, ISG-15, and IRF-3 (p< 0.05) as well as IFN-1 (p

Bin Yuan - One of the best experts on this subject based on the ideXlab platform.

  • the e3 ubiquitin ligase trim21 negatively regulates the innate immune response to intracellular double stranded dna
    Nature Immunology, 2013
    Co-Authors: Zhiqiang Zhang, Ning Lu, Leiyun Weng, Bin Yuan
    Abstract:

    DDX41 is a sensor of intracellular double-stranded DNA (dsDNA) in myeloid dendritic cells (mDCs) that triggers a type I interferon response via the signaling adaptor STING. We identified the E3 ligase TRIM21 as a DDX41-interacting protein and found that knockdown of or deficiency in TRIM21 resulted in enhanced type I interferon responses to intracellular dsDNA and DNA viruses. Overexpression of TRIM21 resulted in more degradation of DDX41 and less production of interferon-β (IFN-β) in response to intracellular dsDNA. The SPRY-PRY domain of TRIM21 interacted with the DEADc domain of DDX41. Lys9 and Lys115 of DDX41 were the targets of TRIM21-mediated ubiquitination. TRIM21 is therefore an interferon-inducible E3 ligase that induces the Lys48 (K48)-linked ubiquitination and degradation of DDX41 and negatively regulates the innate immune response to intracellular dsDNA.

  • the e3 ubiquitin ligase trim21 negatively regulates the innate immune response to intracellular double stranded dna
    Nature Immunology, 2013
    Co-Authors: Zhiqiang Zhang, Ning Lu, Leiyun Weng, Bin Yuan
    Abstract:

    The intracellular sensor DDX41 is important for generating innate responses to DNA viruses. Liu et al. demonstrate that the ubiquitin ligase TRIM21 degrades and thereby regulates DDX41-dependent responses.

  • Corrigendum: The helicase DDX41 senses intracellular DNA mediated by the adaptor STING in dendritic cells
    Nature Immunology, 2012
    Co-Authors: Zhiqiang Zhang, Taeil Kim, Musheng Bao, Bin Yuan, Yong-jun Liu
    Abstract:

    Corrigendum: The helicase DDX41 senses intracellular DNA mediated by the adaptor STING in dendritic cells

  • The helicase DDX41 senses intracellular DNA mediated by the adaptor STING in dendritic cells
    Nature Immunology, 2011
    Co-Authors: Zhiqiang Zhang, Taeil Kim, Musheng Bao, Bin Yuan, Yong-jun Liu
    Abstract:

    The recognition of pathogenic DNA is important to the initiation of antiviral responses. Here we report the identification of DDX41, a member of the DEXDc family of helicases, as an intracellular DNA sensor in myeloid dendritic cells (mDCs). Knockdown of DDX41 expression by short hairpin RNA blocked the ability of mDCs to mount type I interferon and cytokine responses to DNA and DNA viruses. Overexpression of both DDX41 and the membrane-associated adaptor STING together had a synergistic effect in promoting Ifnb promoter activity. DDX41 bound both DNA and STING and localized together with STING in the cytosol. Knockdown of DDX41 expression blocked activation of the mitogen-activated protein kinase TBK1 and the transcription factors NF-κB and IRF3 by B-form DNA. Our results suggest that DDX41 is an additional DNA sensor that depends on STING to sense pathogenic DNA. Sensors of cytosolic nucleic acid can detect the presence of viruses. Liu and colleagues identify the helicase DDX41 as a sensor of double-stranded DNA that initiates upregulation of type I interferon dependent on the adaptor STING in myeloid DCs.

  • the helicase DDX41 senses intracellular dna mediated by the adaptor sting in dendritic cells
    Nature Immunology, 2011
    Co-Authors: Zhiqiang Zhang, Taeil Kim, Musheng Bao, Bin Yuan, Yong-jun Liu
    Abstract:

    The recognition of pathogenic DNA is important to the initiation of antiviral responses. Here we report the identification of DDX41, a member of the DEXDc family of helicases, as an intracellular DNA sensor in myeloid dendritic cells (mDCs). Knockdown of DDX41 expression by short hairpin RNA blocked the ability of mDCs to mount type I interferon and cytokine responses to DNA and DNA viruses. Overexpression of both DDX41 and the membrane-associated adaptor STING together had a synergistic effect in promoting Ifnb promoter activity. DDX41 bound both DNA and STING and localized together with STING in the cytosol. Knockdown of DDX41 expression blocked activation of the mitogen-activated protein kinase TBK1 and the transcription factors NF-κB and IRF3 by B-form DNA. Our results suggest that DDX41 is an additional DNA sensor that depends on STING to sense pathogenic DNA.

Zhiqiang Zhang - One of the best experts on this subject based on the ideXlab platform.

  • DDX41 recognizes bacterial secondary messengers cyclic di gmp and cyclic di amp to activate a type i interferon immune response p1375
    Journal of Immunology, 2013
    Co-Authors: Kislay Parvatiyar, Zhiqiang Zhang, Yan Jiang, Songying Ouyang, Rosane M B Teles, Shankar S Iyer, Shivam A Zaver, Mirjam Schenk, Shang Zeng, Wenwan Zhong
    Abstract:

    Cytosolic detection of bacterially derived secondary messengers cyclic-di-GMP (c-di-GMP) or cyclic -di-AMP (c-di-AMP) by the host immune system activates an innate immune response characterized by the induction of type I interferons (IFNs). Induction of IFN by c-di-GMP or c-di-AMP has been shown to be dependent on a stimulator of IFN genes-TANK binding kinase 1-IFN regulatory factor 3 (STING-TBK1-IRF3) signaling axis. Although STING has been shown to interact with c-di-GMP, an upstream sensor of these cyclic dinucleotides is unknown. Here we identify the helicase, DEAD (Asp-Glu-Ala-Asp) box polypeptide 41 (DDX41) as the pattern recognition receptor (PRR) that senses both c-di-GMP and c-di-AMP. DDX41 specifically and directly interacts with c-di-GMP. Knockdown of DDX41 via shRNA in murine or human immune cells inhibits the induction of innate immune genes and results in defective STING, TBK1 and IRF3 activation in response to c-di-GMP or c-di-AMP. Our findings suggest a mechanism whereby c-di-GMP and c-di-AMP molecules are detected by the DDX41 PRR, which complexes with the STING adaptor to signal to TBK1-IRF3 and activate the IFN response.

  • the e3 ubiquitin ligase trim21 negatively regulates the innate immune response to intracellular double stranded dna
    Nature Immunology, 2013
    Co-Authors: Zhiqiang Zhang, Ning Lu, Leiyun Weng, Bin Yuan
    Abstract:

    DDX41 is a sensor of intracellular double-stranded DNA (dsDNA) in myeloid dendritic cells (mDCs) that triggers a type I interferon response via the signaling adaptor STING. We identified the E3 ligase TRIM21 as a DDX41-interacting protein and found that knockdown of or deficiency in TRIM21 resulted in enhanced type I interferon responses to intracellular dsDNA and DNA viruses. Overexpression of TRIM21 resulted in more degradation of DDX41 and less production of interferon-β (IFN-β) in response to intracellular dsDNA. The SPRY-PRY domain of TRIM21 interacted with the DEADc domain of DDX41. Lys9 and Lys115 of DDX41 were the targets of TRIM21-mediated ubiquitination. TRIM21 is therefore an interferon-inducible E3 ligase that induces the Lys48 (K48)-linked ubiquitination and degradation of DDX41 and negatively regulates the innate immune response to intracellular dsDNA.

  • the e3 ubiquitin ligase trim21 negatively regulates the innate immune response to intracellular double stranded dna
    Nature Immunology, 2013
    Co-Authors: Zhiqiang Zhang, Ning Lu, Leiyun Weng, Bin Yuan
    Abstract:

    The intracellular sensor DDX41 is important for generating innate responses to DNA viruses. Liu et al. demonstrate that the ubiquitin ligase TRIM21 degrades and thereby regulates DDX41-dependent responses.

  • the helicase DDX41 recognizes the bacterial secondary messengers cyclic di gmp and cyclic di amp to activate a type i interferon immune response
    Nature Immunology, 2012
    Co-Authors: Kislay Parvatiyar, Zhiqiang Zhang, Yan Jiang, Songying Ouyang, Rosane M B Teles, Shankar S Iyer, Shivam A Zaver, Mirjam Schenk, Shang Zeng, Wenwan Zhong
    Abstract:

    The induction of type I interferons by the bacterial secondary messengers cyclic di-GMP (c-di-GMP) or cyclic di-AMP (c-di-AMP) is dependent on a signaling axis that involves the adaptor STING, the kinase TBK1 and the transcription factor IRF3. Here we identified the heliase DDX41 as a pattern-recognition receptor (PRR) that sensed both c-di-GMP and c-di-AMP. DDX41 specifically and directly interacted with c-di-GMP. Knockdown of DDX41 via short hairpin RNA in mouse or human cells inhibited the induction of genes encoding molecules involved in the innate immune response and resulted in defective activation of STING, TBK1 and IRF3 in response to c-di-GMP or c-di-AMP. Our results suggest a mechanism whereby c-di-GMP and c-di-AMP are detected by DDX41, which forms a complex with STING to signal to TBK1-IRF3 and activate the interferon response.

  • Corrigendum: The helicase DDX41 senses intracellular DNA mediated by the adaptor STING in dendritic cells
    Nature Immunology, 2012
    Co-Authors: Zhiqiang Zhang, Taeil Kim, Musheng Bao, Bin Yuan, Yong-jun Liu
    Abstract:

    Corrigendum: The helicase DDX41 senses intracellular DNA mediated by the adaptor STING in dendritic cells

Jassy Mary S. Lazarte - One of the best experts on this subject based on the ideXlab platform.

  • Immunostimulatory effect of DDX41 of olive flounder (Paralichthys olivaceus)
    Food and Agricultural Immunology, 2017
    Co-Authors: Jassy Mary S. Lazarte, Young-rim Kim, Jung Seok Lee, Si Won Kim, Jae Wook Jung, Jaesung Kim, Jeong-ho Lee, Tae Sung Jung
    Abstract:

    ABSTRACTThe cellular DEAD-box helicase DDX41, which functions as an initial sensor for cytoplasmic DNA, is involved in the activation of type I interferon (IFN-1) immune response in olive flounder. A plasmid encoding DDX41 (pEF-D) was introduced into flounder cells in vitro and in vivo. Immune responses induced by DDX41 were evaluated by relative quantification value (ΔΔCt) method of RT-qPCR using specific IFN-related gene primers. Results in in vitro, transcript levels of IFN-1, IRF-3, ISG-15 and IL-1β were significantly higher in pEF-D- than pEF-A-transfected cells, with 15-, 4-, 10- and 32-fold changes, respectively. In vivo, elevated expression of these genes was observed in the kidney of pEF-D group on days 1 and 3 post-treatment. The viral challenge test revealed higher survival rate in fish treated with pEF-D (67.5%) than controls, PBS- and pEF-A-treated fish (45%). Conclusively DDX41 elicits a robust IFN-mediated immune response, validating its adjuvant property.

  • enhancement of glycoprotein based dna vaccine for viral hemorrhagic septicemia virus vhsv via addition of the molecular adjuvant DDX41
    Fish & Shellfish Immunology, 2017
    Co-Authors: Jassy Mary S. Lazarte, Young-rim Kim, Jung Seok Lee, Si Won Kim, Jae Wook Jung, Jaesung Kim, Woo Jai Lee, Tae Sung Jung
    Abstract:

    The use of molecular adjuvants to improve the immunogenicity of DNA vaccines has been thoroughly studied in recent years. Glycoprotein (G)-based DNA vaccines had been proven to be effective in combating infection against Rhabdovirus (especially infectious hematopoietic necrosis virus, IHNV) in salmonids. DDX41 is a helicase known to induce antiviral and inflammatory responses by inducing a type I IFN innate immune response. To gain more information regarding G-based DNA vaccines in olive flounder (Paralicthys olivaceus), we tried to develop a more efficient G-based DNA vaccine by adding a molecular adjuvant, DDX41. We designed a DNA vaccine in which the VHSV glycoprotein (G-protein) and DDX41 were driven by the EF-1α and CMV promoters, respectively. Olive flounders were intramuscularly immunized with 1 μg of plasmids encoding the G-based DNA vaccine alone (pEF-G), the molecular adjuvant alone (pEF-D), or the vaccine-adjuvant construct (pEF-GD). At two different time points, 15 and 30 days later, the fish were intraperitoneally infected with VHSV (100 μL; 1 × 106 TCID50/mL). Our assays revealed that the plasmid constructs showed up-regulated expression of IFN-1 and its associated genes at day 3 post-vaccination in both kidney and spleen samples. Specifically, pEF-GD showed statistically higher expression of immune response genes than pEF-G and pEF-D treated group (p < 0.05/p < 0.001). After VHSV challenge, the fish group treated with pEF-GD showed higher survival rate than the pEF-G treated group, though difference was not statistically significant in the 15 dpv challenged group however in the 30 dpv challenged group, the difference was statistically significant (p < 0.05). Together, these results clearly demonstrate that DDX41 is an effective adjuvant for the G-based DNA vaccine in olive flounder. Our novel findings could facilitate the development of more effective DNA vaccines for the aquaculture industry.

  • Enhancement of glycoprotein-based DNA vaccine for viral hemorrhagic septicemia virus (VHSV) via addition of the molecular adjuvant, DDX41.
    Fish & shellfish immunology, 2017
    Co-Authors: Jassy Mary S. Lazarte, Young-rim Kim, Jung Seok Lee, Si Won Kim, Jae Wook Jung, Jaesung Kim, Woo Jai Lee, Tae Sung Jung
    Abstract:

    The use of molecular adjuvants to improve the immunogenicity of DNA vaccines has been thoroughly studied in recent years. Glycoprotein (G)-based DNA vaccines had been proven to be effective in combating infection against Rhabdovirus (especially infectious hematopoietic necrosis virus, IHNV) in salmonids. DDX41 is a helicase known to induce antiviral and inflammatory responses by inducing a type I IFN innate immune response. To gain more information regarding G-based DNA vaccines in olive flounder (Paralicthys olivaceus), we tried to develop a more efficient G-based DNA vaccine by adding a molecular adjuvant, DDX41. We designed a DNA vaccine in which the VHSV glycoprotein (G-protein) and DDX41 were driven by the EF-1α and CMV promoters, respectively. Olive flounders were intramuscularly immunized with 1 μg of plasmids encoding the G-based DNA vaccine alone (pEF-G), the molecular adjuvant alone (pEF-D), or the vaccine-adjuvant construct (pEF-GD). At two different time points, 15 and 30 days later, the fish were intraperitoneally infected with VHSV (100 μL; 1 × 106 TCID50/mL). Our assays revealed that the plasmid constructs showed up-regulated expression of IFN-1 and its associated genes at day 3 post-vaccination in both kidney and spleen samples. Specifically, pEF-GD showed statistically higher expression of immune response genes than pEF-G and pEF-D treated group (p 

  • enhancement of immunogenicity in glycoprotein based dna vaccine by the addition of DDX41 a molecular adjuvant
    Fish & Shellfish Immunology, 2016
    Co-Authors: Jassy Mary S. Lazarte, Young-rim Kim, Jung Seok Lee, Si Won Kim, Jae Wook Jung, Jaesung Kim, Se Pyeong Lim, Tae Sung Jung
    Abstract:

    The emergence of the first commercialized DNA vaccine for fish had prompted for studies that are focused on the development of a more efficacious vaccine that can protect cultured fish against virus infection. This DNA vaccine designed to express viral glycoprotein (G) gene was proven to effectively give protection to several species of fish against rhabdoviruses (ex. viral hemorrhagic septicemia virus, VHSV). Thus, the addition of molecular adjuvant to the DNA vaccine has been considered to be one of the best methods in improving the vaccine’s capability since its presence triggers a more enhanced immune response. DDX41, a cytosolic sensor, has the ability to activate an antiviral cascade of events in response to stimulation of a DNA virus making it a good molecular adjuvant candidate. In the present study, we designed a DNA vaccine consisting of VHSV glycoprotein (VHSVg) and DDX41 which is regulated by EF-1α and CMV promoters, respectively. The aim is to develop a DNA vaccine that can elicit a stronger immune response that could protect the fish against VHSV infection. Four plasmid constructs were prepared for the experiment: pEF-A (empty vector), pEF-G (with VHSVg), pEF-D (with DDX41) and pEF-GD (both VHSVg and DDX41). Expression of individual genes was checked using Western Blot assay, VHSVg in pEF-G and DDX41 in pEF-D were detected using V5 antibody while DDX41 in pEF-GD was detected using myc antibody. Several immune response genes were also evaluated through qPCR to further elucidate the antiviral effect of the plasmid constructs. ∆∆Ct method was utilized to quantify the fold changes of the immune gene transcripts after immunization. qPCR results showed that there are significant increases in the transcript number of IRF-1, ISG-15, and IRF-3 (p< 0.05) as well as IFN-1 (p<0.1) in cells that were immunized with the vaccine-adjuvant construct, pEF-GD, compared to the control plasmid, pEF-A. The results implicated the strength of DDX41 in inducing IFN-mediated immune responses which demonstrates its ability as a molecular adjuvant. This is essential since majority of the DNA vaccines against fish rhabdovirus that we have now are purely based on viral glycoprotein and though it has been proven to be effective, the development of a more efficient DNA vaccine that can improve fish immunity against rhabdovirus infection is significantly beneficial to the aquaculture industry.

  • Enhancement of immunogenicity in glycoprotein-based DNA vaccine by the addition of DDX41, a molecular adjuvant
    Fish & Shellfish Immunology, 2016
    Co-Authors: Jassy Mary S. Lazarte, Young-rim Kim, Jung Seok Lee, Si Won Kim, Jae Wook Jung, Jaesung Kim, Pyeong Lim, Tae Sung Jung
    Abstract:

    The emergence of the first commercialized DNA vaccine for fish had prompted for studies that are focused on the development of a more efficacious vaccine that can protect cultured fish against virus infection. This DNA vaccine designed to express viral glycoprotein (G) gene was proven to effectively give protection to several species of fish against rhabdoviruses (ex. viral hemorrhagic septicemia virus, VHSV). Thus, the addition of molecular adjuvant to the DNA vaccine has been considered to be one of the best methods in improving the vaccine’s capability since its presence triggers a more enhanced immune response. DDX41, a cytosolic sensor, has the ability to activate an antiviral cascade of events in response to stimulation of a DNA virus making it a good molecular adjuvant candidate. In the present study, we designed a DNA vaccine consisting of VHSV glycoprotein (VHSVg) and DDX41 which is regulated by EF-1α and CMV promoters, respectively. The aim is to develop a DNA vaccine that can elicit a stronger immune response that could protect the fish against VHSV infection. Four plasmid constructs were prepared for the experiment: pEF-A (empty vector), pEF-G (with VHSVg), pEF-D (with DDX41) and pEF-GD (both VHSVg and DDX41). Expression of individual genes was checked using Western Blot assay, VHSVg in pEF-G and DDX41 in pEF-D were detected using V5 antibody while DDX41 in pEF-GD was detected using myc antibody. Several immune response genes were also evaluated through qPCR to further elucidate the antiviral effect of the plasmid constructs. ∆∆Ct method was utilized to quantify the fold changes of the immune gene transcripts after immunization. qPCR results showed that there are significant increases in the transcript number of IRF-1, ISG-15, and IRF-3 (p< 0.05) as well as IFN-1 (p

Young-rim Kim - One of the best experts on this subject based on the ideXlab platform.

  • Immunostimulatory effect of DDX41 of olive flounder (Paralichthys olivaceus)
    Food and Agricultural Immunology, 2017
    Co-Authors: Jassy Mary S. Lazarte, Young-rim Kim, Jung Seok Lee, Si Won Kim, Jae Wook Jung, Jaesung Kim, Jeong-ho Lee, Tae Sung Jung
    Abstract:

    ABSTRACTThe cellular DEAD-box helicase DDX41, which functions as an initial sensor for cytoplasmic DNA, is involved in the activation of type I interferon (IFN-1) immune response in olive flounder. A plasmid encoding DDX41 (pEF-D) was introduced into flounder cells in vitro and in vivo. Immune responses induced by DDX41 were evaluated by relative quantification value (ΔΔCt) method of RT-qPCR using specific IFN-related gene primers. Results in in vitro, transcript levels of IFN-1, IRF-3, ISG-15 and IL-1β were significantly higher in pEF-D- than pEF-A-transfected cells, with 15-, 4-, 10- and 32-fold changes, respectively. In vivo, elevated expression of these genes was observed in the kidney of pEF-D group on days 1 and 3 post-treatment. The viral challenge test revealed higher survival rate in fish treated with pEF-D (67.5%) than controls, PBS- and pEF-A-treated fish (45%). Conclusively DDX41 elicits a robust IFN-mediated immune response, validating its adjuvant property.

  • enhancement of glycoprotein based dna vaccine for viral hemorrhagic septicemia virus vhsv via addition of the molecular adjuvant DDX41
    Fish & Shellfish Immunology, 2017
    Co-Authors: Jassy Mary S. Lazarte, Young-rim Kim, Jung Seok Lee, Si Won Kim, Jae Wook Jung, Jaesung Kim, Woo Jai Lee, Tae Sung Jung
    Abstract:

    The use of molecular adjuvants to improve the immunogenicity of DNA vaccines has been thoroughly studied in recent years. Glycoprotein (G)-based DNA vaccines had been proven to be effective in combating infection against Rhabdovirus (especially infectious hematopoietic necrosis virus, IHNV) in salmonids. DDX41 is a helicase known to induce antiviral and inflammatory responses by inducing a type I IFN innate immune response. To gain more information regarding G-based DNA vaccines in olive flounder (Paralicthys olivaceus), we tried to develop a more efficient G-based DNA vaccine by adding a molecular adjuvant, DDX41. We designed a DNA vaccine in which the VHSV glycoprotein (G-protein) and DDX41 were driven by the EF-1α and CMV promoters, respectively. Olive flounders were intramuscularly immunized with 1 μg of plasmids encoding the G-based DNA vaccine alone (pEF-G), the molecular adjuvant alone (pEF-D), or the vaccine-adjuvant construct (pEF-GD). At two different time points, 15 and 30 days later, the fish were intraperitoneally infected with VHSV (100 μL; 1 × 106 TCID50/mL). Our assays revealed that the plasmid constructs showed up-regulated expression of IFN-1 and its associated genes at day 3 post-vaccination in both kidney and spleen samples. Specifically, pEF-GD showed statistically higher expression of immune response genes than pEF-G and pEF-D treated group (p < 0.05/p < 0.001). After VHSV challenge, the fish group treated with pEF-GD showed higher survival rate than the pEF-G treated group, though difference was not statistically significant in the 15 dpv challenged group however in the 30 dpv challenged group, the difference was statistically significant (p < 0.05). Together, these results clearly demonstrate that DDX41 is an effective adjuvant for the G-based DNA vaccine in olive flounder. Our novel findings could facilitate the development of more effective DNA vaccines for the aquaculture industry.

  • Enhancement of glycoprotein-based DNA vaccine for viral hemorrhagic septicemia virus (VHSV) via addition of the molecular adjuvant, DDX41.
    Fish & shellfish immunology, 2017
    Co-Authors: Jassy Mary S. Lazarte, Young-rim Kim, Jung Seok Lee, Si Won Kim, Jae Wook Jung, Jaesung Kim, Woo Jai Lee, Tae Sung Jung
    Abstract:

    The use of molecular adjuvants to improve the immunogenicity of DNA vaccines has been thoroughly studied in recent years. Glycoprotein (G)-based DNA vaccines had been proven to be effective in combating infection against Rhabdovirus (especially infectious hematopoietic necrosis virus, IHNV) in salmonids. DDX41 is a helicase known to induce antiviral and inflammatory responses by inducing a type I IFN innate immune response. To gain more information regarding G-based DNA vaccines in olive flounder (Paralicthys olivaceus), we tried to develop a more efficient G-based DNA vaccine by adding a molecular adjuvant, DDX41. We designed a DNA vaccine in which the VHSV glycoprotein (G-protein) and DDX41 were driven by the EF-1α and CMV promoters, respectively. Olive flounders were intramuscularly immunized with 1 μg of plasmids encoding the G-based DNA vaccine alone (pEF-G), the molecular adjuvant alone (pEF-D), or the vaccine-adjuvant construct (pEF-GD). At two different time points, 15 and 30 days later, the fish were intraperitoneally infected with VHSV (100 μL; 1 × 106 TCID50/mL). Our assays revealed that the plasmid constructs showed up-regulated expression of IFN-1 and its associated genes at day 3 post-vaccination in both kidney and spleen samples. Specifically, pEF-GD showed statistically higher expression of immune response genes than pEF-G and pEF-D treated group (p 

  • enhancement of immunogenicity in glycoprotein based dna vaccine by the addition of DDX41 a molecular adjuvant
    Fish & Shellfish Immunology, 2016
    Co-Authors: Jassy Mary S. Lazarte, Young-rim Kim, Jung Seok Lee, Si Won Kim, Jae Wook Jung, Jaesung Kim, Se Pyeong Lim, Tae Sung Jung
    Abstract:

    The emergence of the first commercialized DNA vaccine for fish had prompted for studies that are focused on the development of a more efficacious vaccine that can protect cultured fish against virus infection. This DNA vaccine designed to express viral glycoprotein (G) gene was proven to effectively give protection to several species of fish against rhabdoviruses (ex. viral hemorrhagic septicemia virus, VHSV). Thus, the addition of molecular adjuvant to the DNA vaccine has been considered to be one of the best methods in improving the vaccine’s capability since its presence triggers a more enhanced immune response. DDX41, a cytosolic sensor, has the ability to activate an antiviral cascade of events in response to stimulation of a DNA virus making it a good molecular adjuvant candidate. In the present study, we designed a DNA vaccine consisting of VHSV glycoprotein (VHSVg) and DDX41 which is regulated by EF-1α and CMV promoters, respectively. The aim is to develop a DNA vaccine that can elicit a stronger immune response that could protect the fish against VHSV infection. Four plasmid constructs were prepared for the experiment: pEF-A (empty vector), pEF-G (with VHSVg), pEF-D (with DDX41) and pEF-GD (both VHSVg and DDX41). Expression of individual genes was checked using Western Blot assay, VHSVg in pEF-G and DDX41 in pEF-D were detected using V5 antibody while DDX41 in pEF-GD was detected using myc antibody. Several immune response genes were also evaluated through qPCR to further elucidate the antiviral effect of the plasmid constructs. ∆∆Ct method was utilized to quantify the fold changes of the immune gene transcripts after immunization. qPCR results showed that there are significant increases in the transcript number of IRF-1, ISG-15, and IRF-3 (p< 0.05) as well as IFN-1 (p<0.1) in cells that were immunized with the vaccine-adjuvant construct, pEF-GD, compared to the control plasmid, pEF-A. The results implicated the strength of DDX41 in inducing IFN-mediated immune responses which demonstrates its ability as a molecular adjuvant. This is essential since majority of the DNA vaccines against fish rhabdovirus that we have now are purely based on viral glycoprotein and though it has been proven to be effective, the development of a more efficient DNA vaccine that can improve fish immunity against rhabdovirus infection is significantly beneficial to the aquaculture industry.

  • Enhancement of immunogenicity in glycoprotein-based DNA vaccine by the addition of DDX41, a molecular adjuvant
    Fish & Shellfish Immunology, 2016
    Co-Authors: Jassy Mary S. Lazarte, Young-rim Kim, Jung Seok Lee, Si Won Kim, Jae Wook Jung, Jaesung Kim, Pyeong Lim, Tae Sung Jung
    Abstract:

    The emergence of the first commercialized DNA vaccine for fish had prompted for studies that are focused on the development of a more efficacious vaccine that can protect cultured fish against virus infection. This DNA vaccine designed to express viral glycoprotein (G) gene was proven to effectively give protection to several species of fish against rhabdoviruses (ex. viral hemorrhagic septicemia virus, VHSV). Thus, the addition of molecular adjuvant to the DNA vaccine has been considered to be one of the best methods in improving the vaccine’s capability since its presence triggers a more enhanced immune response. DDX41, a cytosolic sensor, has the ability to activate an antiviral cascade of events in response to stimulation of a DNA virus making it a good molecular adjuvant candidate. In the present study, we designed a DNA vaccine consisting of VHSV glycoprotein (VHSVg) and DDX41 which is regulated by EF-1α and CMV promoters, respectively. The aim is to develop a DNA vaccine that can elicit a stronger immune response that could protect the fish against VHSV infection. Four plasmid constructs were prepared for the experiment: pEF-A (empty vector), pEF-G (with VHSVg), pEF-D (with DDX41) and pEF-GD (both VHSVg and DDX41). Expression of individual genes was checked using Western Blot assay, VHSVg in pEF-G and DDX41 in pEF-D were detected using V5 antibody while DDX41 in pEF-GD was detected using myc antibody. Several immune response genes were also evaluated through qPCR to further elucidate the antiviral effect of the plasmid constructs. ∆∆Ct method was utilized to quantify the fold changes of the immune gene transcripts after immunization. qPCR results showed that there are significant increases in the transcript number of IRF-1, ISG-15, and IRF-3 (p< 0.05) as well as IFN-1 (p