Characterization of transgenic mice containing adenovirus early region 3 genomic DNA

Fejér, György; Győry, Ildikó; Tufariello, Jo Ann M.; Horwitz, Marshall S.

doi:10.1128/jvi.68.9.5871-5881.1994

Cited by 17 publications

(4 citation statements)

References 44 publications

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“…We found expression of the early viral E1A gene in spleen and liver ( Fig. S1B), but not in lung and kidney (not shown), which agrees with earlier findings on in vivo Ad tropism [47,48]. Expression of IFN-a and IFN-b mRNA was below the level of detection in the organs of non-infected controls.…”

Section: Splenic Myeloid Dendritic Cells Are the Major Producers Of Isupporting

confidence: 92%

Key Role of Splenic Myeloid DCs in the IFN-αβ Response to Adenoviruses In Vivo

et al. 2008

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The early systemic production of interferon (IFN)-αβ is an essential component of the antiviral host defense mechanisms, but is also thought to contribute to the toxic side effects accompanying gene therapy with adenoviral vectors. Here we investigated the IFN-αβ response to human adenoviruses (Ads) in mice. By comparing the responses of normal, myeloid (m)DC- and plasmacytoid (p)DC-depleted mice and by measuring IFN-αβ mRNA expression in different organs and cells types, we show that in vivo, Ads elicit strong and rapid IFN-αβ production, almost exclusively in splenic mDCs. Using knockout mice, various strains of Ads (wild type, mutant and UV-inactivated) and MAP kinase inhibitors, we demonstrate that the Ad-induced IFN-αβ response does not require Toll-like receptors (TLR), known cytosolic sensors of RNA (RIG-I/MDA-5) and DNA (DAI) recognition and interferon regulatory factor (IRF)-3, but is dependent on viral endosomal escape, signaling via the MAP kinase SAPK/JNK and IRF-7. Furthermore, we show that Ads induce IFN-αβ and IL-6 in vivo by distinct pathways and confirm that IFN-αβ positively regulates the IL-6 response. Finally, by measuring TNF-α responses to LPS in Ad-infected wild type and IFN-αβR−/− mice, we show that IFN-αβ is the key mediator of Ad-induced hypersensitivity to LPS. These findings indicate that, like endosomal TLR signaling in pDCs, TLR-independent virus recognition in splenic mDCs can also produce a robust early IFN-αβ response, which is responsible for the bulk of IFN-αβ production induced by adenovirus in vivo. The signaling requirements are different from known TLR-dependent or cytosolic IFN-αβ induction mechanisms and suggest a novel cytosolic viral induction pathway. The hypersensitivity to components of the microbial flora and invading pathogens may in part explain the toxic side effects of adenoviral gene therapy and contribute to the pathogenesis of adenoviral disease.

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Section: Splenic Myeloid Dendritic Cells Are the Major Producers Of Isupporting

confidence: 92%

Key Role of Splenic Myeloid DCs in the IFN-αβ Response to Adenoviruses In Vivo

et al. 2008

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“…GFP transduction was assessed by fluorescence and phase-contrast microscopy or fluorescence-activated cell sorter (FACS) analysis. The viruses were propagated and purified by CsCl 2 density gradient ultracentrifugation as previously described ( 57 ). The concentration of virus particles was determined with a spectrophotometer.…”

Section: Methodsmentioning

confidence: 99%

Key Role of the Scavenger Receptor MARCO in Mediating Adenovirus Infection and Subsequent Innate Responses of Macrophages

Maler

Nielsen

Stichling

et al. 2017

mBio

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The scavenger receptor MARCO is expressed in several subsets of naive tissue-resident macrophages and has been shown to participate in the recognition of various bacterial pathogens. However, the role of MARCO in antiviral defense is largely unexplored. Here, we investigated whether MARCO might be involved in the innate sensing of infection with adenovirus and recombinant adenoviral vectors by macrophages, which elicit vigorous immune responses in vivo. Using cells derived from mice, we show that adenovirus infection is significantly more efficient in MARCO-positive alveolar macrophages (AMs) and in AM-like primary macrophage lines (Max Planck Institute cells) than in MARCO-negative bone marrow-derived macrophages. Using antibodies blocking ligand binding to MARCO, as well as gene-deficient and MARCO-transfected cells, we show that MARCO mediates the rapid adenovirus transduction of macrophages. By enhancing adenovirus infection, MARCO contributes to efficient innate virus recognition through the cytoplasmic DNA sensor cGAS. This leads to strong proinflammatory responses, including the production of interleukin-6 (IL-6), alpha/beta interferon, and mature IL-1α. These findings contribute to the understanding of viral pathogenesis in macrophages and may open new possibilities for the development of tools to influence the outcome of infection with adenovirus or adenovirus vectors.

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“…The relative abundance of all spliced mRNAs and thus the relative protein levels of MAV-1 E3 may be dependent on the tissue or cell type. Cell-specific splicing control has been seen in transgenic mice expressing the human adenovirus E3 (20). It is possible that, during MAV-1 infections of mice, each class of E3 protein is more abundantly expressed in a certain tissue type where it plays a specific role.…”

Section: Discussionmentioning

confidence: 99%

Analysis of early region 3 mutants of mouse adenovirus type 1

Beard

Spindler

1996

J Virol

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Early region 3 (E3) of mouse adenovirus type 1 has the potential to produce three proteins which have identical amino termini but unique carboxy-terminal sequences. Three recombinant deletion viruses were constructed so that each could produce only one of the three E3 proteins. A fourth mutant that should produce no E3 proteins was also constructed. These recombinants were able to grow in mouse 3T6 cells and produced wild-type levels of viral mRNAs and proteins except for those specifically deleted by the mutations. Early mRNA production from the mutant viruses was analyzed by reverse transcriptase PCR and confirmed that each deletion mutant would be able to produce only one of the three E3 proteins. Late mRNA production was analyzed by Northern (RNA) blotting and found to be similar in wild-type and mutant viruses. Capsid morphology was unaltered in the mutant viruses as seen by electron microscopy. Immunoprecipitation of E3 proteins from infections of mouse 3T6 cells using an antiserum specific for all three E3 proteins was used to examine the effect of the introduced mutations on protein expression. Two mutants produced only one class of E3 protein as predicted from their specific mutations and mRNA expression profiles. One mutant virus failed to produce any detectable E3 proteins. The predicted E3-null mutant was found to be leaky and could produce low levels of E3 proteins. Outbred Swiss mice were infected with the E3 mutant viruses to determine if the E3 proteins have an effect on the pathogenicity of the virus in mice. All of the mutants showed decreased pathogenicity as determined by increased 50% lethal doses, indicating that the proteins of the E3 region are important determinants of the pathogenesis of mouse adenovirus in its natural host.

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Characterization of transgenic mice containing adenovirus early region 3 genomic DNA

Cited by 17 publications

References 44 publications

Key Role of Splenic Myeloid DCs in the IFN-αβ Response to Adenoviruses In Vivo

Key Role of Splenic Myeloid DCs in the IFN-αβ Response to Adenoviruses In Vivo

Key Role of the Scavenger Receptor MARCO in Mediating Adenovirus Infection and Subsequent Innate Responses of Macrophages

Analysis of early region 3 mutants of mouse adenovirus type 1

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