Cytomegalovirus infection elicits a conserved chemokine response from human and guinea pig amnion cells

Putri, Dira S.; Berkebile, Zachary W.; Mustafa, Hiba J.; Fernández-Alarcón, Claudia; Abrahante, Juan E.; Schleiss, Mark R.; Bierle, Craig J.

doi:10.1016/j.virol.2020.06.005

Cited by 5 publications

(10 citation statements)

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“…Curiously, while the concentration of CXCL10 in amniotic fluid correlates with CMV genome abundance, the chemokine does not appear to accumulate in amniotic fluid when other viruses cause intrauterine infections (32). We and others have found that CXCL10 transcription is upregulated when placental cells or tissue are infected with HCMV or GPCMV in vitro (46,84,85). Indoleamine-2,3-dioxygenase (IDO) is an enzyme that is encoded by two genes, IDO1 and IDO2, that catalyze the ratelimiting step in tryptophan catabolism.…”

Section: Discussionmentioning

confidence: 87%

“…This study found that GPCMV infects both the amnion and the yolk sack, that viral loads in the two membranes are correlated, and that the highest rates of infection and viral loads in the membranes occur after maternal infection late in pregnancy (46). While GPCMV joins HCMV on a short list of viruses that have been observed to infect the fetal membranes, it remains unclear whether or how viral infections of the fetal membrane affect the fetus (24,73).…”

Section: Discussionmentioning

confidence: 90%

“…A minimally tissue culture adapted stock of GPCMV 22122 (ATCC VR-682), prepared as previously described by passaging guinea pig salivary gland homogenate twice on JH4 guinea pig lung fibroblasts (ATCC CCL-158), was used for this study ( 33 , 45 , 46 ). JH4 cells were purchased from ATCC and propagated according to their specifications excepting that the growth media was supplemented with sodium bicarbonate ( 47 ).…”

Section: Methodsmentioning

confidence: 99%

“…After DNA was extracted from whole blood, tissue, and amniotic fluid, GPCMV genomes were quantified by droplet digital PCR (ddPCR) using primers and probes specific to GP54 using the Bio-Rad QX200 system as previously described ( 46 ). ddPCR results were analyzed using the QuantaSoft™ Analysis Pro software (Bio-Rad); GPCMV viral loads are presented as the number of copies genome per ml of fluid or mg of tissue.…”

Section: Methodsmentioning

confidence: 99%

“…RNA was extracted from placenta as described above. A two-step reverse transcriptase ddPCR (RT-ddPCR) protocol was used to quantify transcript abundance ( 46 ). cDNA was synthesized from total RNA using the Maxima™ H Minus cDNA Synthesis Master Mix (ThermoFisher).…”

Section: Methodsmentioning

confidence: 99%

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The Placental Response to Guinea Pig Cytomegalovirus Depends Upon the Timing of Maternal Infection

et al. 2021

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Human cytomegalovirus (HCMV) infects the placenta, and these placental infections can cause fetal injury and/or demise. The timing of maternal HCMV infection during pregnancy is a determinant of fetal outcomes, but how development affects the placenta’s susceptibility to infection, the likelihood of placental injury post-infection, and the frequency of transplacental HCMV transmission remains unclear. In this study, guinea pig cytomegalovirus (GPCMV) was used to model primary maternal infection and compare the effects of infection at two different times on the placenta. When guinea pigs were infected with GPCMV at either 21- or 35-days gestation (dGA), maternal and placental viral loads, as determined by droplet digital PCR, were not significantly affected by the timing of maternal infection. However, when the transcriptomes of gestational age-matched GPCMV-infected and control placentas were compared, significant infection-associated changes in gene expression were only observed after maternal infection at 35 dGA. Notably, transcripts associated with immune activation (e.g. Cxcl10, Ido1, Tgtp1, and Tlr8) were upregulated in the infected placenta. A GPCMV-specific in situ hybridization assay detected rare infected cells in the main placenta after maternal infection at either time, and maternal infection at 35 dGA also caused large areas of GPCMV-infected cells in the junctional zone. As GPCMV infection after mid-gestation is known to cause high rates of stillbirth and/or fetal growth restriction, our results suggest that the placenta becomes sensitized to infection-associated injury late in gestation, conferring an increased risk of adverse pregnancy outcomes after cytomegalovirus infection.

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Section: Discussionmentioning

confidence: 87%

Section: Discussionmentioning

confidence: 90%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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The Placental Response to Guinea Pig Cytomegalovirus Depends Upon the Timing of Maternal Infection

et al. 2021

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Human Trophoblast Stem Cells Restrict Human Cytomegalovirus Replication

Rollman,

Berkebile,

Okae

et al. 2023

Preprint

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Placental infection plays a central role in the pathogenesis of congenital human cytomegalovirus (HCMV) infections and is a cause of fetal growth restriction and pregnancy loss. HCMV can replicate in some trophoblast cell types, but it remains unclear how the virus evades antiviral immunity in the placenta and how infection compromises placental development and function. Human trophoblast stem cells (TSCs) can be differentiated into extravillous trophoblasts (EVTs), syncytiotrophoblasts (STBs), and organoids, and this study assessed the utility of TSCs as a model of HCMV infection in the first trimester placenta. HCMV was found to non-productively infect TSCs, EVTs, and STBs. Immunofluorescence assays and flow cytometry experiments further revealed that infected TSCs frequently only express immediate early viral gene products. Similarly, RNA-sequencing found that viral gene expression in TSCs does not follow the kinetic patterns observed during lytic infection in fibroblasts. Canonical antiviral responses were largely not observed in HCMV-infected TSCs and TSC-derived trophoblasts. Rather, infection dysregulated factors involved in cell identity, differentiation, and WNT signaling. Thus, while HCMV does not replicate in TSCs, infection may perturb trophoblast differentiation in ways that could interfere with placental function.ImportancePlacental infection plays a central role in HCMV pathogenesis during pregnancy, but the species-specificity of HCMV and the limited availability and lifespan of primary trophoblasts have been persistent barriers to understanding how infection impacts this vital organ. Human TSCs represent a new approach to modeling viral infection early in placental development. This study reveals that TSCs, like other stem cell types, restrict HCMV replication. However, infection perturbs the expression of genes involved in differentiation and cell fate determination, pointing to a mechanism by which HCMV could cause placental injury.

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CD4⁺but not CD8⁺T cells are required for protection against severe guinea pig cytomegalovirus infections

Rollman,

Berkebile,

Hicks

et al. 2024

Preprint

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Human cytomegalovirus (HCMV) is a ubiquitous herpesvirus and the leading cause of infectious disease related birth defects worldwide. How the immune response modulates the risk of intrauterine transmission of HCMV after maternal infection remains poorly understood. Maternal T cells likely play a critical role in preventing infection at the maternal-fetal interface and limiting spread across the placenta, but concerns exist that immune responses to infection may also cause placental dysfunction and adverse pregnancy outcomes. This study investigated the role of CD4+and CD8+T cells in a guinea pig model of primary cytomegalovirus infection. Monoclonal antibodies specific to guinea pig CD4 and CD8 were used to deplete T cells in non- pregnant and in pregnant guinea pigs after mid-gestation. CD4+T cell depletion increased the severity of illness, caused significantly elevated viral loads, and increased the rate of congenital guinea pig cytomegalovirus (GPCMV) infection relative to animals treated with control antibody. CD8+T cell depletion was comparably well tolerated and did not significantly affect the weight of infected guinea pigs or viral loads in their blood or tissue. However, significantly more viral genomes and transcripts were detected in the placenta and decidua of CD8+T cell depleted dams post-infection. This study corroborates earlier findings made in nonhuman primates that maternal CD4+T cells play a critical role in limiting the severity of primary CMV infection during pregnancy while also revealing that other innate and adaptive immune responses can compensate for an absent CD8+T cell response in α-CD8-treated guinea pigs.Author SummaryCongenital cytomegalovirus infection is a leading cause of adverse pregnancy outcomes and preventable disability in children. Using guinea pigs, a well-established small animal model of congenital infection and intrauterine development, this study tested how depleting T cells affects the course of primary cytomegalovirus infections. Severe illness and high rates of congenital infection were observed when helper CD4+T cells were depleted. The depletion of killer CD8+T cells did not affect the severity of disease or the rate of congenital infection but did increase the amount of virus that was detected in the placenta. A greater understanding how an immune response can prevent the infection of the placenta and developing offspring is needed to inform vaccine and therapeutic development. This study not only describes a new reagent that can be used to study the guinea pig immune system but also sheds new light in how adaptive immunity regulates congenital viral infection.

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Cytomegalovirus infection elicits a conserved chemokine response from human and guinea pig amnion cells

Cited by 5 publications

References 57 publications

The Placental Response to Guinea Pig Cytomegalovirus Depends Upon the Timing of Maternal Infection

The Placental Response to Guinea Pig Cytomegalovirus Depends Upon the Timing of Maternal Infection

Human Trophoblast Stem Cells Restrict Human Cytomegalovirus Replication

CD4⁺but not CD8⁺T cells are required for protection against severe guinea pig cytomegalovirus infections

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Cytomegalovirus infection elicits a conserved chemokine response from human and guinea pig amnion cells

Cited by 5 publications

References 57 publications

The Placental Response to Guinea Pig Cytomegalovirus Depends Upon the Timing of Maternal Infection

The Placental Response to Guinea Pig Cytomegalovirus Depends Upon the Timing of Maternal Infection

Human Trophoblast Stem Cells Restrict Human Cytomegalovirus Replication

CD4+but not CD8+T cells are required for protection against severe guinea pig cytomegalovirus infections

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Product

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CD4⁺but not CD8⁺T cells are required for protection against severe guinea pig cytomegalovirus infections