Innate Immune Response Induced by Baculovirus Attenuates Transgene Expression in Mammalian Cells

Ono, Chikako; Ninomiya, Ai; Yamamoto, Seiji; Abe, Takayuki; Wen, Xiauyu; Fukuhara, Takasuke; Sasai, Miwa; Yamamoto, Masahiro; Saitoh, Tatsuya; Satoh, Takashi; Kawai, Taro; Ishii, Ken J.; Akira, Shizuo; Okamoto, Tomoyoshi; Matsuura, Yoshiharu

doi:10.1128/jvi.03055-13

Cited by 29 publications

(46 citation statements)

References 82 publications

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“…To establish another BHK-T7 cell line (BHK-T7/P5), BSR cells (3), a derivative of BHK cells, were selected by transfection with a eukaryotic expression plasmid encoding T7pol under the control of a strong CMV early enhancer/chicken β-actin (CAG) promoter (38), followed by incubation in the presence of 4 μg/mL puromycin (Sigma-Aldrich). Immortalized MEFs derived from TBK1 +/− IKKi −/− (TBK1 +/− ) and TBK1 −/− IKKi −/− (TBK1 −/− ) mice were prepared as previously described (39,40) and grown in DMEM supplemented to contain 10% FBS, 100 units/mL penicillin, and 100 μg/mL streptomycin. Human cancer cell lines, HT29, PC3, HCC-2998, and OVCAR-4, were cultured in RPMI1640 (Nacalai Tesque) supplemented to contain 10% FBS, 100 units/mL penicillin, and 100 μg/mL streptomycin.…”

Section: Methodsmentioning

confidence: 99%

Entirely plasmid-based reverse genetics system for rotaviruses

Kanai

Komoto

Kawagishi

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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194

271

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Rotaviruses (RVs) are highly important pathogens that cause severe diarrhea among infants and young children worldwide. The understanding of the molecular mechanisms underlying RV replication and pathogenesis has been hampered by the lack of an entirely plasmid-based reverse genetics system. In this study, we describe the recovery of recombinant RVs entirely from cloned cDNAs. The strategy requires coexpression of a small transmembrane protein that accelerates cell-to-cell fusion and vaccinia virus capping enzyme. We used this system to obtain insights into the process by which RV nonstructural protein NSP1 subverts host innate immune responses. By insertion into the NSP1 gene segment, we recovered recombinant viruses that encode split-green fluorescent protein-tagged NSP1 and NanoLuc luciferase. This technology will provide opportunities for studying RV biology and foster development of RV vaccines and therapeutics.

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Section: Methodsmentioning

confidence: 99%

Entirely plasmid-based reverse genetics system for rotaviruses

Kanai

Komoto

Kawagishi

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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194

271

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“…Knockdown of Rab2b or Garil5 mRNA inhibited transcription of Ifnb1 and Cxcl10 induced by modified vaccinia virus Ankara strain (MVA) (Figures 6A and 6B). Knockdown of Rab2b or Garil5 mRNA also inhibited production of IFN-β induced by baculovirus, which triggers the cGAS-STING signaling axis (Figure S4; Ono et al, 2014). However, knockdown of Rab2b or Garil5 mRNA did not affect transcription of Ifnb1 and Cxcl10 induced by encephalomyocarditis virus (EMCV), an RNA virus that is recognized by MDA5 (Figures 6A and 6B; Kato et al, 2006).…”

Section: Resultsmentioning

confidence: 96%

“…Modified vaccinia virus Ankara strain was purchased from the American Type Culture Collection. EMCV and baculovirus has been described previously (Kato et al, 2006; Ono et al, 2014). The viral titers were determined with 50% tissue culture infectious dose (TCID 50 ) assays as described previously (Saitoh et al, 2009).…”

Section: Methodsmentioning

confidence: 99%

The RAB2B-GARIL5 Complex Promotes Cytosolic DNA-Induced Innate Immune Responses

et al. 2017

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Summary Cyclic GMP-AMP synthase (cGAS) is a cytosolic DNA sensor that induces the IFN antiviral response. However, the regulatory mechanisms that mediate cGAS-triggered signaling have not been fully explored. Here, we show the involvement of a small GTPase RAB2B and its effector protein Golgi-associated RAB2B interactor-like 5 (GARIL5) in the cGAS-mediated IFN response. RAB2B-deficiency affects the IFN response induced by cytosolic DNA. Consistent with this, RAB2B-deficiency enhances replication of vaccinia virus, a DNA virus. After DNA stimulation, RAB2B colocalizes with stimulator of interferon genes (STING), the downstream signal mediator of cGAS, on the Golgi apparatus. GTP-binding activity of RAB2B is required for its localization on the Golgi apparatus and for recruitment of GARIL5. GARIL5 deficiency also affects the IFN response induced by cytosolic DNA and enhances replication of vaccinia virus. These findings indicate that the RAB2B-GARIL5 complex promotes IFN responses against DNA viruses by regulating the cGAS-STING signaling axis.

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“…106 In transduced cells and to a lesser extent in APCs, free baculoviral DNA released into the cytosol triggers the production of type-I IFNs through RIG-I/IPS-1/TBK1/IRF3 and cGAS/STING/TBK1/IRF3 pathways, resulting in the suppression of transgene expression. 145 Actually, the transient gene expression in transduced mammalian cells discouraged the use of baculoviral vectors for gene therapy, which was the initial focus for BacMam vectors. 146 Immunization using BacMam vectors has been widely reported, as these vectors can transduce mammalian cells in vivo, facilitating a transient expression of the gene of interest and the development of Ag-specific immune responses.…”

Section: Innate Sensing Of Baculoviruses Results In Adaptive Immune Rmentioning

confidence: 99%

Induction of mucosal immunity against pathogens by using recombinant baculoviral vectors: Mechanisms, advantages, and limitations

Fragoso-Saavedra

Vega-Lopez

2020

Journal of Leukocyte Biology

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Over 90% of pathogens of medical importance invade the organism through mucosal surfaces, which makes it urgent to develop safe and effective mucosal vaccines and mucosal immunization protocols. Besides, parenteral immunization does not provide adequate protective immunity in mucosal surfaces. Effective mucosal vaccination could protect local and systemic compartments and favor herd immunity. Although various mucosal adjuvants and Ag‐delivery systems have been developed, none has filled the gap to control diseases caused by complex mucosal pathogens. Among the strategies to counteract them, recombinant virions from the baculovirus Autographa californica multiple nucleopolyhedrovirus (rAcMNPV) are useful vectors, given their safety and efficacy to produce mucosal and systemic immunity in animal infection models. Here, we review the immunogenic properties of rAcMNPV virions from the perspectives of mucosal immunology and vaccinology. Some features, which are analyzed and extrapolated from studies with different particulate antigens, include size, shape, surface molecule organization, and danger signals, all needed to break the tolerogenic responses of the mucosal immune tissues. Also, we present a condensed discussion on the immunity provided by rAcMNPV virions against influenza virus and human papillomavirus in animal models. Through the text, we highlight the advantages and limitations of this experimental immunization platform.

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Innate Immune Response Induced by Baculovirus Attenuates Transgene Expression in Mammalian Cells

Cited by 29 publications

References 82 publications

Entirely plasmid-based reverse genetics system for rotaviruses

Entirely plasmid-based reverse genetics system for rotaviruses

The RAB2B-GARIL5 Complex Promotes Cytosolic DNA-Induced Innate Immune Responses

Induction of mucosal immunity against pathogens by using recombinant baculoviral vectors: Mechanisms, advantages, and limitations

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