Protein coding content of the ULb′ region of wild-type rhesus cytomegalovirus

Oxford, Kristie; Eberhardt, Meghan K.; Yang, Kai; Strelow, Lisa; Kelly, S.; Zhou, Shan; Barry, Peter A.

doi:10.1016/j.virol.2007.10.040

Cited by 55 publications

(98 citation statements)

References 26 publications

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“…UL141 mediates NK cell immune evasion by downregulating CD155 and CD112, ligands of the NK cell receptors DNAM-1 (CD226) and TACTILE (CD96) (57,73). The UL141 homologue Rh164 is absent in RhCMV 180.92 due to genomic rearrangements during fibroblast adaptation (51,64). Whether any of the point mutations found in the BAC-derived RhCMV genome affects protein function remains to be determined.…”

Section: Resultsmentioning

confidence: 99%

“…Two different strains of RhCMV, 68-1 and 180.92, have been fully sequenced previously (31,64), and partial sequences of various regions of the genome from low-passage-number isolates have been published (51). However, many of the in vivo experiments performed to date, particularly those involving recombinant CMVs, employed RhCMV strain 68-1 cloned as a self-excis-able bacterial artificial chromosome (BAC) (12).…”

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confidence: 99%

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Reevaluation of the Coding Potential and Proteomic Analysis of the BAC-Derived Rhesus Cytomegalovirus Strain 68-1

Malouli

Nakayasu

Viswanathan

et al. 2012

J Virol

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Cytomegaloviruses are highly host restricted, resulting in cospeciation with their hosts. As a natural pathogen of rhesus macaques (RM), rhesus cytomegalovirus (RhCMV) has therefore emerged as a highly relevant experimental model for pathogenesis and vaccine development due to its close evolutionary relationship to human CMV (HCMV). Most in vivo experiments performed with RhCMV employed strain 68-1 cloned as a bacterial artificial chromosome (BAC). However, the complete genome sequence of the 68-1 BAC has not been determined. Furthermore, the gene content of the RhCMV genome is unknown, and previous open reading frame (ORF) predictions relied solely on uninterrupted ORFs with an arbitrary cutoff of 300 bp. To obtain a more precise picture of the actual proteins encoded by the most commonly used molecular clone of RhCMV, we reevaluated the RhCMV 68-1 BAC genome by whole-genome shotgun sequencing and determined the protein content of the resulting RhCMV virions by proteomics. By comparing the RhCMV genome to those of several related Old World monkey (OWM) CMVs, we were able to filter out many unlikely ORFs and obtain a simplified map of the RhCMV genome. This comparative genomics analysis suggests a high degree of ORF conservation among OWM CMVs, thus decreasing the likelihood that ORFs found only in RhCMV comprise true genes. Moreover, virion proteomics independently validated the revised ORF predictions, since only proteins that were conserved across OWM CMVs could be detected. Taken together, these data suggest a much higher conservation of genome and virion structure between CMVs of humans, apes, and OWMs than previously assumed.

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Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

Reevaluation of the Coding Potential and Proteomic Analysis of the BAC-Derived Rhesus Cytomegalovirus Strain 68-1

Malouli

Nakayasu

Viswanathan

et al. 2012

J Virol

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“…The parental strain used to generate Δpp65ab, RhCMV 68-1, shows reduced secretion from infected animals, most likely due to the lack of genes in the ULb′-homology region required for tissue tropism (56). Since RhCMV 68-1 does not generate robust plasma viremia in infected animals, the appearance of RhCMV-Δpp65ab in plasma samples became particularly striking.…”

Section: Discussionmentioning

confidence: 99%

Cytomegalovirus pp65 limits dissemination but is dispensable for persistence

et al. 2014

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The most abundantly produced virion protein in human cytomegalovirus (HCMV) is the immunodominant phosphoprotein 65 (pp65), which is frequently included in CMV vaccines. Although it is nonessential for in vitro CMV growth, pp65 displays immunomodulatory functions that support a potential role in primary and/or persistent infection. To determine the contribution of pp65 to CMV infection and immunity, we generated a rhesus CMV lacking both pp65 orthologs (RhCMVΔpp65ab). While deletion of pp65ab slightly reduced growth in vitro and increased defective particle formation, the protein composition of secreted virions was largely unchanged. Interestingly, pp65 was not required for primary and persistent infection in animals. Immune responses induced by RhCMVΔpp65ab did not prevent reinfection with rhesus CMV; however, reinfection with RhCMVΔUS2-11, which lacks viral-encoded MHC-I antigen presentation inhibitors, was prevented. Unexpectedly, induction of pp65b-specific T cells alone did not protect against RhCMVΔUS2-11 challenge, suggesting that T cells targeting multiple CMV antigens are required for protection. However, pp65-specific immunity was crucial for controlling viral dissemination during primary infection, as indicated by the marked increase of RhCMVΔpp65ab genome copies in CMV-naive, but not CMV-immune, animals. Our data provide rationale for inclusion of pp65 into CMV vaccines but also demonstrate that pp65-induced T cell responses alone do not recapitulate the protective effect of natural infection.

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“…A statistically significant association of CMI and oral shedding was observed only in group 3 (P ϭ 0.015, 1 degree of freedom, F ϭ 16.9). terms of both genetic coding content (32), particularly within UL/bЈ, and the capacity for high-level, persistent shedding in bodily fluids, such as saliva (5,23). Coupled with the genetic drift within the gB coding region between the vaccine antigen and the challenge virus, this study design more closely represents the obstacles facing HCMV vaccine trials than those of previous animal studies.…”

Section: Discussionmentioning

confidence: 99%

“…All of the monkeys were challenged by a subcutaneous route with 1 ϫ 10 5 PFU of RhCMV variant UCD52, which is an epithelial (26,32). RhCMV UCD52 is pathogenic in inoculated rhesus fetuses (43).…”

Section: Methodsmentioning

confidence: 99%

Vaccine-Induced Control of Viral Shedding following Rhesus Cytomegalovirus Challenge in Rhesus Macaques

Abel

Martinez

Yue

et al. 2011

J Virol

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The use of animal models of human cytomegalovirus (HCMV) infection is critical to refine HCMV vaccine candidates. Previous reports have demonstrated that immunization of rhesus monkeys against rhesus cytomegalovirus (RhCMV) can reduce both local and systemic replication of RhCMV following experimental RhCMV challenge. These studies used prime/boost combinations of DNA expression plasmids alone or DNA priming and boosting with either inactivated virion particles or modified vaccinia virus Ankara (MVA) expressing the same antigens. Viral outcomes included reduced RhCMV replication at the site of subcutaneous inoculation and RhCMV viremia following intravenous inoculation. Since shedding of cytomegalovirus from mucosal surfaces is critical for horizontal transmission of the virus, DNA priming/MVA boosting was evaluated for the ability to reduce oral shedding of RhCMV following subcutaneous challenge. Of six rhesus monkeys vaccinated exclusively against RhCMV glycoprotein B (gB), phosphoprotein 65 (pp65), and immediate-early 1 (IE1), half showed viral loads in saliva that were lower than those of control monkeys by 1 to 3 orders of magnitude. Further, there was a strong association of memory pp65 T cell responses postchallenge in animals exhibiting the greatest reduction in oral shedding. These results highlight the fact that a DNA/MVA vaccination regimen can achieve a notable reduction in a critical parameter of viral replication postchallenge. The recently completed clinical trial of a gB subunit vaccine in which the rate of HCMV infection was reduced by 50% in the individuals receiving the vaccine is consistent with the results of this study suggesting that additional immunogens are likely essential for maximum protection in an outbred human population.The nearly 40-year quest for a vaccine that confers protective efficacy against congenital infection with human cytomegalovirus (HCMV) remains unmet, although considerable progress has been made. Complexities in HCMV's natural history, incompletely defined correlates of immune protection, and financial and logistical factors in designing sufficiently powered clinical trials all contribute to the absence of a licensed HCMV vaccine(s). Animal model studies with rhesus monkey, mouse, and guinea pig systems have demonstrated that multiple vaccine strategies, including approaches based on those proposed for HCMV, are effective at limiting the extent of challenge virus replication. Immunization of HCMV-negative women with recombinant gB has been clinically evaluated (50,51). A recently completed phase II trial assessed the efficacy of the vaccine to decrease cases of maternal HCMV infection (34). The endpoint of this study was the time to HCMV infection (50, 51), and the trial ended earlier than planned because vaccine efficacy exceeded goals. The results offer strong encouragement that vaccination regimes directed at prominent neutralizing epitopes can significantly decrease the rate of primary infection in HCMV-negative women. The impressive, but less than 100%, level ...

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Protein coding content of the ULb′ region of wild-type rhesus cytomegalovirus

Cited by 55 publications

References 26 publications

Reevaluation of the Coding Potential and Proteomic Analysis of the BAC-Derived Rhesus Cytomegalovirus Strain 68-1

Reevaluation of the Coding Potential and Proteomic Analysis of the BAC-Derived Rhesus Cytomegalovirus Strain 68-1

Cytomegalovirus pp65 limits dissemination but is dispensable for persistence

Vaccine-Induced Control of Viral Shedding following Rhesus Cytomegalovirus Challenge in Rhesus Macaques

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