Modeling of Human Cytomegalovirus Maternal-Fetal Transmission in a Novel Decidual Organ Culture

Weisblum, Yiska; Panet, Amos; Zakay‐Rones, Zichria; Haimov‐Kochman, Ronit; Goldman‐Wohl, Debra; Ariel, Ilana; Falk, Haya; Natanson-Yaron, Shira; Goldberg, Miri D.; Gilad, Ronit; Lurain, Nell S.; Greenfield, Caryn; Wolf, Dana

doi:10.1128/jvi.05749-11

Cited by 69 publications

(84 citation statements)

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“…(D) Neurospheres at day 11 post-induction of differentiation were infected with the low-passage-number clinical strain CI5006, recovered from the urine of a congenitally infected newborn. Shown are the viral IE-1 and late RNA R160461 mRNA levels, determined as described previously (27) at the indicated times postinfection and normalized by the housekeeping gene for glucose-6-phosphate dehydrogenase (G6PD) (similar results were obtained with three additional low-passage-number clinical isolates).…”

supporting

confidence: 59%

“…A similar lack of pp65 internalization in hESC was observed at sequential time points between 2 and 48 h postinfection (viral incubation times equaled inspection times for these experiments; data not shown). (C and D) Quantitative viral DNA measurements, performed as described previously (27) in the indicated cell lysates, normalized by RNase P (RnP) gene DNA copies. (C) Viral entry assay.…”

mentioning

confidence: 99%

“…1A) as detailed previously (24, 25). The emerging neural spheres were infected at sequential time points along the neural differentiation process with the broadly tropic HCMV TB40/E strain expressing green fluorescent protein (GFP) (26,27) (Fig. 1B).…”

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Transition toward Human Cytomegalovirus Susceptibility in Early Human Embryonic Stem Cell-Derived Neural Precursors

Berger

Gil

Panet

et al. 2015

J Virol

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Congenital human cytomegalovirus (HCMV) infection is associated with neurodevelopmental disabilities. To dissect the earliest events of infection in the developing human brain, we studied HCMV infection during controlled differentiation of human embryonic stem cells (hESC) into neural precursors. We traced a transition from viral restriction in hESC, mediated by a block in viral binding, toward HCMV susceptibility in early hESC-derived neural precursors. We further revealed the role of plateletderived growth factor receptor alpha (PDGFR␣) as a determinant of the developmentally acquired HCMV susceptibility. Human cytomegalovirus (HCMV) is a leading cause of congenital infection (1), associated with neurodevelopmental disabilities (2, 3). The ability of the virus to infect the developing fetal brain is a key factor in its neuropathogenesis (3)(4)(5)(6)(7)(8). While considerable experimental data were obtained from newborn and embryonic mouse models (6, 9-15), the strict species specificity precludes animal models of HCMV. Recent studies in human neuronal progenitor cells (NPC) derived from fetal and neonatal brains have revealed productive HCMV infection of NPC, with resultant functional alterations (16)(17)(18)(19)(20). Notably, NPC do not represent early neural embryonic development but rather a more advanced stage along the neuronal differentiation route. Hence, the earliest events defining the susceptibility of the developing human nervous system to HCMV have remained largely unknown.Human embryonic stem cells (hESC) provide an opportunity to study early human neural development (21-23). We have developed highly reproducible protocols for controlled induction of differentiation of hESC toward a neural lineage, giving rise to enriched populations of proliferating, developmentally multipotent early NPC (24,25).Here, we have employed experimental HCMV infection in a dynamic model of controlled differentiation of hESC into neural precursors, to gain insight into the molecular events mediating HCMV infection during early human neurodevelopment.We first sought to track the earliest stage of neural differentiation at which the cells become susceptible to HCMV. To this end, differentiation of hESC was induced in floating cell clusters toward the formation of neural spheres (Fig. 1A) as detailed previously (24, 25). The emerging neural spheres were infected at sequential time points along the neural differentiation process with the broadly tropic HCMV TB40/E strain expressing green fluorescent protein (GFP) (26,27) (Fig. 1B). In accordance with previous studies (25), immunostaining analysis for the pluripotent stem cell marker TRA 1-81 and the neural differentiation marker PSA-NCAM showed that the majority of the cells underwent early neural differentiation within 11 days (Fig. 1C). Importantly, we have identified a transition toward HCMV susceptibility occurring between 4 and 11 days post-differentiation induction (Fig. 1B). A similar susceptibility pattern was observed for low-passage-number, cell-associated, clinic...

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confidence: 99%

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Transition toward Human Cytomegalovirus Susceptibility in Early Human Embryonic Stem Cell-Derived Neural Precursors

Berger

Gil

Panet

et al. 2015

J Virol

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“…Preclinical cell culture models continue to provide evidence that antibodies prevent infection, 85,86 which is an important backdrop to vaccine strategies aiming to mimic or improve on natural immunity. Specifically, the predominance of gH pentamer-specific antibodies in immune sera combined with a broader understanding of cell types that support initial infection, dissemination and transplacental transmission have spawned efforts to improve the response directed at the gH pentamer complex as well as efforts to improve gB-specific antibody titers to more efficiently neutralize viruses in epithelial and endothelial cells.…”

Section: Cmv-specific Protective Immune Response Parametersmentioning

confidence: 99%

The immunological underpinnings of vaccinations to prevent cytomegalovirus disease

McCormick

Mocarski

2014

Cell Mol Immunol

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A universal cytomegalovirus (CMV) vaccination promises to reduce the burden of the developmental damage that afflicts up to 0.5% of live births worldwide. An effective vaccination that prevents transplacental transmission would reduce CMV congenital disease and CMV-associated still births and leave populations less susceptible to opportunistic CMV disease. Thus, a vaccination against this virus has long been recognized for the potential of enormous health-care savings because congenital damage is life-long and existing anti-viral options are limited. Vaccine researchers, industry leaders, and regulatory representatives have discussed the challenges posed by clinical efficacy trials that would lead to a universal CMV vaccine, reviewing the links between infection and disease, and identifying settings where disrupting viral transmission might provide a surrogate endpoint for disease prevention. Reducing the complexity of such trials would facilitate vaccine development. Children and adolescents are the targets for universal vaccination, with the expectation of protecting the offspring of immunized women. Given that a majority of females worldwide experience CMV infection during childhood, a universal vaccine must boost natural immunity and reduce transmission due to reactivation and re-infection as well as primary infection during pregnancy. Although current vaccine strategies recognize the value of humoral and cellular immunity, the precise mechanisms that act at the placental interface remain elusive. Immunity resulting from natural infection appears to limit rather than prevent reactivation of latent viruses and susceptibility to re-infection, leaving a challenge for universal vaccination to improve upon natural immunity levels. Despite these hurdles, early phase clinical trials have achieved primary end points in CMV seronegative subjects. Efficacy studies must be expanded to mixed populations of CMV-naive and naturally infected subjects to understand the overall efficacy and potential. Together with CMV vaccine candidates currently in clinical development, additional promising preclinical strategies continue to come forward; however, these face limitations due to the insufficient understanding of host defense mechanisms that prevent transmission, as well as the age-old challenges of reaching the appropriate threshold of immunogenicity, efficacy, durability and potency. This review focuses on the current understanding of natural and CMV vaccine-induced protective immunity.

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“…Initial observations suggest that CMV hyperimmunoglobulin (HIG) could inhibit viral spread in vitro 17,18 , restore placental health in mothers during primary infection 19 and lead to the regression of fetal cerebral ultrasound abnormalities 20 .…”

Section: Passive Immunisationmentioning

confidence: 99%

Vaccination for cytomegalovirus: when, where and how

Dasari

Khanna

2015

Microbiol. Aust.

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Although following primary human cytomegalovirus (CMV)infection in many individuals no overt symptoms are observed, CMV came to medical attention due to its significant morbidity and mortality associated with congenital infec- In pregnant women following natural primary infection, viral replication is controlled by the emergence of antigen-specific CD4 + , CD8 + and CD45RA + effector memory T-cells 7,8 . Lower frequencies of CMV-specific CD4 + T-cells and CD45RA + cells in mothers following primary infection is known to be associated with virus transmission to the fetus 9,10 . Based on these observations it can be hypothesised that emergence of higher frequencies of CMV-specific CD4 + , CD8 + and CD45RA + effector memory T-cells may be associated with control of viremia and prevention of transplacental transmission. Therefore, an effective CMV vaccine specially designed to induce CMV-specific CD4 + , CD8 + and CD45RA + effector memory T-cells in women of reproductive age could potentially decrease CMV transmission to the fetus and vaccination of infants, toddlers and adolescents could reduce the duration of viral shedding, which may reduce child-to-mother transmission.Clinical evaluation of active and passive immunisation strategies against CMV in the context of congenital CMV Live attenuated virus vaccinesThe initial CMV vaccine trials based on an attenuated form of CMV isolates Towne and AD169 11,12 induced humoral and cellular Under the Microscope

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Modeling of Human Cytomegalovirus Maternal-Fetal Transmission in a Novel Decidual Organ Culture

Cited by 69 publications

References 80 publications

Transition toward Human Cytomegalovirus Susceptibility in Early Human Embryonic Stem Cell-Derived Neural Precursors

Transition toward Human Cytomegalovirus Susceptibility in Early Human Embryonic Stem Cell-Derived Neural Precursors

The immunological underpinnings of vaccinations to prevent cytomegalovirus disease

Vaccination for cytomegalovirus: when, where and how

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