Modeling Adenovirus Latency in Human Lymphocyte Cell Lines

Zhang, Yange; Huang, Weijie; Ornelles, David A.; Gooding, Linda R.

doi:10.1128/jvi.00562-10

Cited by 47 publications

(78 citation statements)

References 39 publications

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“…6, HAdV-5 DNA was detectable with our PLP (Fig. 6A), thereby confirming the previous observation that HAdV-5 can infect and replicate in BJAB cells (23). Consequently, E1A (Fig.…”

Section: Detection Of Hadv-5 Dna and Viral Mrnas In Long-term Infectisupporting

confidence: 77%

“…Theoretically, low viral RNA and high DNA content in HeLa cells at 25 hpi may mark the commencement of virus release from the cell, whereas a similar scenario in BJAB cells at 6 dpi might specify the cells where HAdV-5 establishes persistent infections. Notably, Zhang and colleagues (23) have reported that HAdV-5-infected BJAB cells show a drastic reduction in viral hexon protein expression while maintaining relatively high viral DNA amounts during long-term infection.…”

Section: Discussionmentioning

confidence: 99%

“…In contrast to the well-characterized HAdV lytic infections, much less is known about the virus behavior in long-term latent/persistent infections in B and T lymphocytes (23). Therefore, we applied our method to study an HAdV-5 infection in the B lymphocyte cell line, BJAB.…”

Section: Discussionmentioning

confidence: 99%

“…Recent studies have shown that HAdV-5 also can cause long-term latent/persistent infections in B-and T-lymphocyte cell lines (23,33,34). Therefore, it became of interest to test if the RCA method could define viral DNA accumulation and mRNA expression patterns in HAdV-5-infected B lymphocytes.…”

Section: Detection Of Hadv-5 Dna and Viral Mrnas In Long-term Infectimentioning

confidence: 99%

“…For example, infection efficiency of HAdV-5 depends on the cell cycle status and expression of specific viral genes (22). Also, during long-term latent/persistent HAdV-5 infection, the amount of viral DNA does not necessarily correlate with viral gene transcription, as the viral genomes can be transcriptionally silenced in these infections (23). Hence, single-cell studies can be more informative for monitoring viral DNA accumulation and mRNA expression in individual HAdV-infected cells.…”

mentioning

confidence: 99%

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Simultaneous Single-Cell In Situ Analysis of Human Adenovirus Type 5 DNA and mRNA Expression Patterns in Lytic and Persistent Infection

Krzywkowski

Ciftci

Assadian

et al. 2017

J Virol

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An efficient adenovirus infection results in high-level accumulation of viral DNA and mRNAs in the infected cell population. However, the average viral DNA and mRNA content in a heterogeneous cell population does not necessarily reflect the same abundance in individual cells. Here, we describe a novel padlock probe-based rolling-circle amplification technique that enables simultaneous detection and analysis of human adenovirus type 5 (HAdV-5) genomic DNA and virusencoded mRNAs in individual infected cells. We demonstrate that the method is applicable for detection and quantification of HAdV-5 DNA and mRNAs in short-term infections in human epithelial cells and in long-term infections in human B lymphocytes. Single-cell evaluation of these infections revealed high heterogeneity and unique cell subpopulations defined by differential viral DNA content and mRNA expression. Further, our single-cell analysis shows that the specific expression pattern of viral E1A 13S and 12S mRNA splice variants is linked to HAdV-5 DNA content in the individual cells. Furthermore, we show that expression of a mature form of the HAdV-5 histone-like protein VII affects virus genome detection in HAdV-5-infected cells. Collectively, padlock probes combined with rolling-circle amplification should be a welcome addition to the method repertoire for the characterization of the molecular details of the HAdV life cycle in individual infected cells.IMPORTANCE Human adenoviruses (HAdVs) have been extensively used as model systems to study various aspects of eukaryotic gene expression and genome organization. The vast majority of the HAdV studies are based on standard experimental procedures carried out using heterogeneous cell populations, where data averaging often masks biological differences. As every cell is unique, characteristics and efficiency of an HAdV infection can vary from cell to cell. Therefore, the analysis of HAdV gene expression and genome organization would benefit from a method that permits analysis of individual infected cells in the heterogeneous cell population. Here, we show that the padlock probe-based rolling-circle amplification method can be used to study concurrent viral DNA accumulation and mRNA expression patterns in individual HAdV-5-infected cells. Hence, this versatile method can be applied to detect the extent of infection and virus gene expression changes in different HAdV-5 infections.KEYWORDS adenovirus, persistent infection, lytic infection, rolling-circle amplification, single-cell analysis H uman adenoviruses (HAdV) are double-stranded DNA (dsDNA) viruses classified into seven distinct species, A to G, with over 60 species described so far (1). Typically, HAdV infection of epithelial cells causes an efficient cell lysis and release of

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“…6, HAdV-5 DNA was detectable with our PLP (Fig. 6A), thereby confirming the previous observation that HAdV-5 can infect and replicate in BJAB cells (23). Consequently, E1A (Fig.…”

Section: Detection Of Hadv-5 Dna and Viral Mrnas In Long-term Infectisupporting

confidence: 77%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Detection Of Hadv-5 Dna and Viral Mrnas In Long-term Infectimentioning

confidence: 99%

mentioning

confidence: 99%

See 3 more Smart Citations

Simultaneous Single-Cell In Situ Analysis of Human Adenovirus Type 5 DNA and mRNA Expression Patterns in Lytic and Persistent Infection

Krzywkowski

Ciftci

Assadian

et al. 2017

J Virol

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Genomic foundations of evolution and ocular pathogenesis in human adenovirus species D

et al. 2019

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Human adenovirus commonly causes infections of respiratory, gastrointestinal, genitourinary, and ocular surface mucosae. Although most adenovirus eye infections are mild and self-limited, specific viruses within human adenovirus species D are associated with epidemic keratoconjunctivitis (EKC), a severe and highly contagious ocular surface infection, which can lead to chronic and/or recurrent, visually disabling keratitis. In this review, we discuss the links between adenovirus ontogeny, genomics, immune responses, and corneal pathogenesis, for those viruses that cause EKC.

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Epigenetics and the dynamics of chromatin during adenovirus infections

et al. 2019

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Edited by Urs GreberAbbreviations ADP, adenovirus death protein; Ad5 or Ad 12, human adenovirus type 5 or 12, or other type referenced; BHK, baby hamster kidney; BrdU, bromodeoxyuridine; CTCF, CCCTC binding factor; DBS, double stranded break; DDR, DNA damage response; E1A, early 1 A; E4orf3 or E4orf6, early 4 open reading frame 3 or 6; HAdV-A12, human adenovirus subgroup A type 12, or different subgroup or type referenced; HMGB, high-mobility group box; hPaf1, human RNA polymerase II-associated factor 1; ISGs, interferon-stimulated genes; PML, promyelocytic leukemia; snRNP, small nuclear ribonuclear protein; SET or TAF-1, SE translocation or template activating factor; VRCs, viral replication centers.

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Modeling Adenovirus Latency in Human Lymphocyte Cell Lines

Cited by 47 publications

References 39 publications

Simultaneous Single-Cell In Situ Analysis of Human Adenovirus Type 5 DNA and mRNA Expression Patterns in Lytic and Persistent Infection

Simultaneous Single-Cell In Situ Analysis of Human Adenovirus Type 5 DNA and mRNA Expression Patterns in Lytic and Persistent Infection

Genomic foundations of evolution and ocular pathogenesis in human adenovirus species D

Epigenetics and the dynamics of chromatin during adenovirus infections

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