Review
- Adenoviral Vector System: A Comprehensive Overview of Constructions, Therapeutic Applications and Host Responses.
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Anyeseu Park, Jeong Yoon Lee
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J. Microbiol. 2024;62(7):491-509. Published online July 22, 2024
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DOI: https://doi.org/10.1007/s12275-024-00159-4
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Abstract
- Adenoviral vectors are crucial for gene therapy and vaccine development, offering a platform for gene delivery into host cells. Since the discovery of adenoviruses, first-generation vectors with limited capacity have evolved to third-generation vectors flacking viral coding sequences, balancing safety and gene-carrying capacity. The applications of adenoviral vectors for gene therapy and anti-viral treatments have expanded through the use of in vitro ligation and homologous recombination, along with gene editing advancements such as CRISPR-Cas9. Current research aims to maintain the efficacy and safety of adenoviral vectors by addressing challenges such as pre-existing immunity against adenoviral vectors and developing new adenoviral vectors from rare adenovirus types and non-human species. In summary, adenoviral vectors have great potential in gene therapy and vaccine development. Through continuous research and technological advancements, these vectors are expected to lead to the development of safer and more effective treatments.
Journal Article
- In Silico Intensive Analysis for the E4 Gene Evolution of Human Adenovirus Species D.
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Chanhee Lee, Anyeseu Park, Jeong Yoon Lee
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J. Microbiol. 2024;62(5):409-418. Published online April 30, 2024
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DOI: https://doi.org/10.1007/s12275-024-00132-1
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Abstract
- Adenovirus (Ad) is a ubiquitous pathogen capable of infecting a wide range of animals and humans. Human Adenovirus (HAdV) can cause severe infection, particularly in individuals with compromised immune systems. To date, over 110 types of HAdV have been classified into seven species from A to G, with the majority belonging to the human adenovirus species D (HAdV-D). In the HAdV-D, the most significant factor for the creation of new adenovirus types is homologous recombination between viral genes involved in determining the virus tropism or evading immune system of host cells. The E4 gene, consisting of seven Open Reading Frames (ORFs), plays a role in both the regulation of host cell metabolism and the replication of viral genes. Despite long-term studies, the function of each ORF remains unclear. Based on our updated information, ORF2, ORF3, and ORF4 have been identified as regions with relatively high mutations compared to other ORFs in the E4 gene, through the use of in silico comparative analysis. Additionally, we managed to visualize high mutation sections, previously undetectable at the DNA level, through a powerful amino acid sequence analysis tool known as proteotyping. Our research has revealed the involvement of the E4 gene in the evolution of human adenovirus, and has established accurate sequence information of the E4 gene, laying the groundwork for further research.
Review
- Genomic Evolution and Recombination Dynamics of Human Adenovirus D Species: Insights from Comprehensive Bioinformatic Analysis.
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Anyeseu Park, Chanhee Lee, Jeong Yoon Lee
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J. Microbiol. 2024;62(5):393-407. Published online March 7, 2024
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DOI: https://doi.org/10.1007/s12275-024-00112-5
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24
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Abstract
- Human adenoviruses (HAdVs) can infect various epithelial mucosal cells, ultimately causing different symptoms in infected organ systems. With more than 110 types classified into seven species (A-G), HAdV-D species possess the highest number of viruses and are the fastest proliferating. The emergence of new adenovirus types and increased diversity are driven by homologous recombination (HR) between viral genes, primarily in structural elements such as the penton base, hexon and fiber proteins, and the E1 and E3 regions. A comprehensive analysis of the HAdV genome provides valuable insights into the evolution of human adenoviruses and identifies genes that display high variation across the entire genome to determine recombination patterns. Hypervariable regions within genetic sequences correlate with functional characteristics, thus allowing for adaptation to new environments and hosts. Proteotyping of newly emerging and already established adenoviruses allows for prediction of the characteristics of novel viruses. HAdV-D species evolved in a direction that increased diversity through gene recombination. Bioinformatics analysis across the genome, particularly in highly variable regions, allows for the verification or re-evaluation of recombination patterns in both newly introduced and pre-existing viruses, ultimately aiding in tracing various biological traits such as virus tropism and pathogenesis. Our research does not only assist in predicting the emergence of new adenoviruses but also offers critical guidance in regard to identifying potential regulatory factors of homologous recombination hotspots.
Journal Article
- Prevalence of Indigenous Antibiotic‑Resistant Salmonella Isolates and Their Application to Explore a Lytic Phage vB_SalS_KFSSM with an Intra‑Broad Specificity
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Jaein Choe , Su-Hyeon Kim , Ji Min Han , Jong-Hoon Kim , Mi-Sun Kwak , Do-Won Jeong , Mi-Kyung Park
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J. Microbiol. 2023;61(12):1063-1073. Published online January 2, 2024
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DOI: https://doi.org/10.1007/s12275-023-00098-6
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Abstract
- The consumption of fresh produce has led to increase in antibiotic-resistant (AR) Salmonella outbreaks. In this study, indigenous
Salmonella was isolated from a total of two hundred-two samples including fresh produce and agricultural environmental
samples in Korea. After biochemical confirmation using the Indole, Methyl Red, Voges-Proskauer, Citrate tests, presumable
Salmonella isolates were identified by 16S rRNA sequencing. Identified Salmonella isolates were evaluated for antibiotic
susceptibility against twenty-two antibiotics. The specificity and the efficiency of plating (EOP) of vB_SalS_KFSSM were
evaluated against fifty-three bacterial strains. Twenty-five suspected Salmonella were isolated and confirmed by the positive
result
for methyl red and citrate, of which ten were identified as Salmonella spp. through 16S rRNA gene sequencing. Eight
Salmonella isolates (4.0%, n = 8/202) were resistant to at least one antibiotic, among which five were multi-drug resistant. As
a lytic phage against Salmonella spp. CMGS-1, vB_SalS_KFSSM was isolated from cow manure. The phage was observed as
a tailed phage belonging to the class Caudoviricetes. It exhibited an intra-broad specificity against four indigenous AR Salmonella
isolates, two indigenous Salmonella isolates, and five other Salmonella serotypes with great efficiencies (EOP ≥ 0.75).
Thus, this study suggested the potential of vB_SalS_KFSSM to combat indigenous AR Salmonella.
Research Support, Non-U.S. Gov'ts
- Detection of Human Adenoviruses and Enteroviruses in Korean Oysters Using Cell Culture, Integrated Cell Culture-PCR, and Direct PCR
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Yoe-Jin Choo , Sang-Jong Kim
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J. Microbiol. 2006;44(2):162-170.
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DOI: https://doi.org/2369 [pii]
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Abstract
- Oysters are known to be carriers of food-born diseases, but research on viruses in Korean oysters is scarce despite its importance for public health. We therefore tested oysters cultivated in Goheung, Seosan, Chungmu, and Tongyeong, for viral contamination using cell culture and integrated cell culture PCR (ICC-PCR) with Buffalo green monkey kidney (BGMK) and human lung epithelial (A549) cells. Additional screens via PCR, amplifying viral nucleic acids extracted from oysters supplemented our analysis. Our methods found 23.6%, 50.9%, and 89.1% of all oysters to be positive for adenoviruses when cell culture, ICC-PCR, and direct PCR, respectively, was used to conduct the screen. The same methodology identified enteroviruses in 5.45%, 30.9%, and 10.9% of all cases. Most of the detected enteroviruses (81.3%) were similar to poliovirus type 1; the remainder resembled coxsackievirus type A1. A homology search with the adenoviral sequences revealed similarities to adenovirus subgenera C (type 2, 5, and 6), D (type 44), and F (enteric type 40 and 41). Adenovirus-positive samples were more abundant in A549 cells (47.3%) than in BGMK cells (18.2%), while the reverse was true for enteroviruses (21.8% vs. 14.5%). Our data demonstrate that Korean oysters are heavily contaminated with enteric viruses, which is readily detectable via ICC-PCR using a combination of A549 and BGMK cells.
- Enhancement of Gene Delivery to Cancer Cells by a Retargeted Adenovirus
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Kwang Seok Oh , Jeffrey A. Engler , Insil Joung
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J. Microbiol. 2005;43(2):179-182.
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DOI: https://doi.org/2164 [pii]
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Abstract
- The inefficiency of in vivo gene transfer using currently available vectors reflects a major hurdle in cancer gene therapy. Both viral and non-viral approaches that improve gene transfer efficiency have been described, but suffer from a number of limitations. Herein, a fiber-modified adenovirus, carrying the small peptide ligand on the capsid, was tested for the delivery of a transgene to cancer cells. The fiber-modified adenovirus was able to mediate the entry and expression of a [beta]-galactosidase into cancer cells with increased efficiency compared to the unmodified adenovirus. Particularly, the gene transfer efficiency was improved up to 5 times in OVCAR3 cells, an ovarian cancer cell line. Such transduction systems hold promise for delivering genes to transferrin receptor overexpressing cancer cells, and could be used for future cancer gene therapy.
- The invariant region I sequence of the adenovirus serotype 2 DNA polymerase influences template specificity during DNA synthesis
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Houng , In Sil
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J. Microbiol. 1998;36(3):222-230.
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Abstract
- Mutants in highly conserved region I (YGDTDS) of the adenovirus serotyope 2 DNA polymerase (Ad Pol) have been shown previously to be defective in assays for initiation and elongation of adenovirus DNA replication in vitro. A selected subset of these mutants was characterized in a number of assays to determine in more detail the nature of the defect that they cause in Ad Pol. The single amino acid substitution in this sequence motif had no detectable effect on binding either to factors required for viral DNA replication or to Ad DNA origin. However, in the deletion mutant mimicking a similar sequence found in the Klenow fragment and in RNA polymerases, binding to Ad DNA origins was reduced. When the nucleotide and template specificity of partially purified mutant Ad Pol proteins was checked there were no significant differences between mutant and wild-type Ad Pols in RNA polymerase assays both on DNA templates and on RNA templates. In reverse transcription assays, both wild type and mutant Ad Pol were inactive, with the exception of a Gly to Met replacement mutant which mimicked the sequence found in reverse transcriptase; this mutant showed low but reproducible levels of reverse transcriptase activity when compared to wt Ad Pol. Taken together, these results suggest that region I may influence template specificity during DNA synthesis, although the single replacement is not sufficient to convert Ad Pol to a reverse transcriptase or an RNA polymerase. Region I probably acts independently of other regions of Ad Pol that are responsible for DNA binding, since mutations in this sequence did not significantly alter adenovirus DNA origin interactions in gel shift assays.
- Functional Implications in Apoptosis by Interferon Inducible Gene Product 1-8D, the Binding Protein to Adenovirus Preterminal Protein
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Insil Joung , Jeffrey A. Engler
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J. Microbiol. 2003;41(4):295-299.
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Abstract
- Adenovirus (Ad) precursor to the terminal protein (pTP) plays an essential roles in the viral DNA replication. Ad pTP serves as a primer for the synthesis of a new DNA strand during the initiation step of replication. In addition, Ad pTP forms organized spherical replication foci on the nuclear matrix (NM) and anchors the viral genome to the NM. Here we identified the interferon inducible gene product 1-8D (Inid) as a pTP binding protein by using a two-hybrid screen of a HeLa cDNA library. Of the clones obtained in this assay, nine were identical to the Inid, a 13-kDa polypeptide that shares homology with genes 1-8U and Leu-13/9-27, most of which have little known functions. The entire open reading frame (ORF) of Inid was cloned into the tetracycline inducible expression vector in order to determine the biological functions related with adenoviral infection. When Inid was introduced to the cells along with adenoviruses, fifty to sixty percent of Ad-infected cells expressing Inid had rounded morphology, which was suggestive of apoptosis. Results from the terminal deoxynucleotidyl transferase (TdT) and DNA fragmentation assays confirmed that Inid induces apoptosis in Ad-infected or in uninfected cells. The Inid binding to pTP may target the cell for apoptotic destruction as a host defense mechanism against the viral infection.