A Recombinant Rhesus Cytomegalovirus Expressing Enhanced Green Fluorescent Protein Retains the Wild-Type Phenotype and Pathogenicity in Fetal Macaques

Chang, W. L. William; Tarantal, Alice F.; Zhou, Shan; Borowsky, Alexander D.; Barry, Peter A.

doi:10.1128/jvi.76.18.9493-9504.2002

Cited by 63 publications

(56 citation statements)

References 52 publications

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“…The BAC vector was engineered into the 210-bp intergenic region of the RhCMV genome between the US2 polyadenylation signal and the US1 transcriptional start site. Our previous study demonstrated that insertion of the EGFP expression cassette into this region does not alter the pathogenicity of the recombinant virus (12). Telo-RF cultures were transfected with the Cre expression vector pOG231 (32) and infected with RhCMV-EGFP (MOI of 1) 2 days after transfection.…”

Section: Resultsmentioning

confidence: 99%

“…primer pairs PAB509-PAB510 and PAB431-PAB489 (12), which are specific for the Cre and EGFP cassettes, respectively. Plasmid transfection and virus reconstitution.…”

mentioning

confidence: 99%

“…Cytoplasmic RNA was isolated by using an RNeasy Mini Kit (Qiagen) according to the manufacturer's instructions. The synthesis of cDNA and the 3Ј rapid amplification of cDNA ends (RACE) for viral IE2, US1, US2, US3, and rhesus glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were performed as previously described (11,12).…”

mentioning

confidence: 99%

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Cloning of the Full-Length Rhesus Cytomegalovirus Genome as an Infectious and Self-Excisable Bacterial Artificial Chromosome for Analysis of Viral Pathogenesis

Chang

Barry

2003

J Virol

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Section: Resultsmentioning

confidence: 99%

“…primer pairs PAB509-PAB510 and PAB431-PAB489 (12), which are specific for the Cre and EGFP cassettes, respectively. Plasmid transfection and virus reconstitution.…”

mentioning

confidence: 99%

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Cloning of the Full-Length Rhesus Cytomegalovirus Genome as an Infectious and Self-Excisable Bacterial Artificial Chromosome for Analysis of Viral Pathogenesis

Chang

Barry

2003

J Virol

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“…All infections were asymptomatic, similar to primary CMV infection of adult humans. To facilitate detection of CMV both in vivo and in vitro, Peter Barry and colleagues recently developed a recombinant CMV expressing GFP [25] as well as RhCMV strain 68.1 in a bacterial artiWcial chromosome, which greatly facilitates generating recombinant viruses [26]. Importantly, both recombinant viruses retained their ability to infect RM suggesting that molecular manipulations do not reduce RhCMV pathogenicity.…”

Section: Pathogenesis Of Rhcmvmentioning

confidence: 99%

Rhesus CMV: an emerging animal model for human CMV

Powers

Früh

2008

Med Microbiol Immunol

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Human CMV is the predominant infectious cause of congenital birth defects and an opportunistic pathogen in immunosuppressed individuals, including AIDS patients. Most individuals are infected early during their life followed by life-long latent infection. During this latent phase, frequent reactivation and antigen production continue to stimulate the immune system. While the immune response is able to control the virus, it is unable to eradicate it. Moreover, super-infection by diVerent CMV strains has been observed despite a strong immune response. Longterm immune stimulation by CMV has also been implicated in immune senescence and chronic conditions such as atherosclerosis. CMVs are highly species-speciWc and the relatedness of CMV genomes exactly mirrors the relatedness of their hosts. Thus, each CMV species is highly adapted to its respective host species, but is unable to infect other, even closely related hosts. While fascinating from an evolutionary perspective, this host restriction prevents studying HCMV in experimental animals. Exceptions are severely immunocompromised mice, e.g. SCID mice, or SCID/NOD mice, which might allow partial reconstitution of CMV infection in rodents. More practical however, is to study CMVs in their natural host, e.g. murine, rat or guinea pig CMVs. However, while these small animal models have many advantages, such as the availability of inbred animals as well as lower cost, the limited homology of the viral genomes with HCMV limits the functional analysis of homologous gene products. The closest relative to HCMV is chimpanzee CMV (CCMV), but this is not a practical animal model since chimps are a protected species, extremely expensive and of very limited availability. In contrast, rhesus macaques are a more widely used experimental animal species and, while more distant than CCMV, rhesus CMV (RhCMV) contains most of the HCMV gene families thus allowing the study of their role in acute and latent CMV infection. In this review we will discuss the current state of developing RhCMV as a model for HCMV.

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“…2). The elicited immune responses were measured by antigenspeciWc ELISA, an EGFP-Xuorescence intensity reductionneutralizing assay using an EGFP-RhCMV [6], and intracellular cytokine staining (ICS). The ICS was performed as previously described protocols [5] to detect the frequency of CD4+ and CD8+ T cells that express IFN-or TNF-upon re-stimulation with a pool of overlapping peptides (15-mers overlapping by 11 amino acids) comprising full-length IE1 or pp65-2 antigen [26].…”

Section: B)mentioning

confidence: 99%

Evaluation of recombinant modified vaccinia Ankara virus-based rhesus cytomegalovirus vaccines in rhesus macaques

Yue

Wang

Abel

et al. 2008

Med Microbiol Immunol

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A vaccine consisting of rhesus cytomegalovirus (RhCMV) pp65-2, gB and IE1 expressed via modiWed vaccinia Ankara (MVA) was evaluated in rhesus macaques with or without prior priming with expression plasmids for the same antigens. Following two MVA treatments, comparable levels of anti-gB, pp65-2 and neutralizing antibody responses, and pp65-2-and IE1-speciWc cellular immune responses were detected in both vaccinated groups. Similar reductions in plasma peak viral loads were observed in these groups compared to untreated controls. This study demonstrates the immunogenicity and protective eYcacy of rMVA-based RhCMV subunit vaccines in a primate host and warrants further investigation to improve the eYcacy of subunit vaccines against CMV.

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A Recombinant Rhesus Cytomegalovirus Expressing Enhanced Green Fluorescent Protein Retains the Wild-Type Phenotype and Pathogenicity in Fetal Macaques

Cited by 63 publications

References 52 publications

Cloning of the Full-Length Rhesus Cytomegalovirus Genome as an Infectious and Self-Excisable Bacterial Artificial Chromosome for Analysis of Viral Pathogenesis

Cloning of the Full-Length Rhesus Cytomegalovirus Genome as an Infectious and Self-Excisable Bacterial Artificial Chromosome for Analysis of Viral Pathogenesis

Rhesus CMV: an emerging animal model for human CMV

Evaluation of recombinant modified vaccinia Ankara virus-based rhesus cytomegalovirus vaccines in rhesus macaques

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