A High-Affinity Native Human Antibody Neutralizes Human Cytomegalovirus Infection of Diverse Cell Types

Kauvar, Lawrence M.; Liu, Keyi; Park, Minha; DeChene, Neal; Stephenson, Robert A.; Tenorio, Edgar; Ellsworth, Stote L.; Tabata, Takako; Petitt, Matthew; Tsuge, Mitsuru; Fang-Hoover, June; Adler, Stuart P.; Cui, Xiaohong; McVoy, Michael A.; Pereira, Lenore

doi:10.1128/aac.04295-14

Cited by 46 publications

(56 citation statements)

References 89 publications

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“…Our findings that cell-cell spread into epithelial cells is resistant to neutralizing antibodies is in direct contrast with a large number of studies in which focus formation in epithelial cells was found to be highly sensitive to neutralizing antibodies (28,30,(32)(33)(34)39). To resolve this discrepancy, we performed focus expansion assays using the TB40-BAC4 strain of HCMV.…”

Section: -E)contrasting

confidence: 68%

“…This observation has profound ramifications for the development of therapeutic interventions, especially as current vaccine studies are focused on the generation of neutralizing antibodies (21)(22)(23)(24)(25)(26)(27). Several of these antibodies are considerably more potent than Cytotect at neutralizing cell-free passaged HCMV (21,22,33). However, biological activity against cell-free spread may not necessarily correlate with activity against cell-cell spread; it will therefore be important to test the ability of these antibodies to limit cell-cell spread, at clinically relevant concentrations, using WT-HCMV.…”

Section: Discussionmentioning

confidence: 99%

“…Many current vaccine strategies are designed to induce a neutralizing antibody response (21)(22)(23)(24)(25)(26)(27). However, it is unclear whether the transmission of HCMV is sensitive or resistant to neutralization, potentially because many studies have used viruses that spread efficiently via the cellfree route (19,(28)(29)(30)(31)(32)(33)(34)(35)(36). To determine the neutralization sensitivity of Merlin, we tested a panel of antibody preparations.…”

Section: Significancementioning

confidence: 99%

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The pentameric complex drives immunologically covert cell–cell transmission of wild-type human cytomegalovirus

Murrell

Bedford

Ladell

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

110

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Human cytomegalovirus (HCMV) strains that have been passaged in vitro rapidly acquire mutations that impact viral growth. These laboratory-adapted strains of HCMV generally exhibit restricted tropism, produce high levels of cell-free virus, and develop susceptibility to natural killer cells. To permit experimentation with a virus that retained a clinically relevant phenotype, we reconstructed a wild-type (WT) HCMV genome using bacterial artificial chromosome technology. Like clinical virus, this genome proved to be unstable in cell culture; however, propagation of intact virus was achieved by placing the RL13 and UL128 genes under conditional expression. In this study, we show that WT-HCMV produces extremely low titers of cell-free virus but can efficiently infect fibroblasts, epithelial, monocyte-derived dendritic, and Langerhans cells via direct cell-cell transmission. This process of cell-cell transfer required the UL128 locus, but not the RL13 gene, and was significantly less vulnerable to the disruptive effects of IFN, cellular restriction factors, and neutralizing antibodies compared with cell-free entry. Resistance to neutralizing antibodies was dependent on high-level expression of the pentameric gH/gL/gpUL128-131A complex, a feature of WT but not passaged strains of HCMV.virology | immune evasion | herpesvirus | HCMV | cell-cell spread H uman cytomegalovirus (HCMV) is a major cause of morbidity and mortality in the immunocompromised and the leading infectious cause of congenital malformation. As a result, a vaccine has been designated as a priority. However, basic studies of clinically relevant isolates that inform our understanding of the disease process are limited due to the rapid accumulation of genetic mutations during in vitro passage of HCMV. The same three genetic sites are reproducibly affected: the RL13 gene; the UL128 locus (UL128L), which comprises UL128, UL30, and UL131A; and the ∼15-kb U L /b′ gene region (1). Deletions in the U L /b′ region can affect tropism [UL148 (2)], latency [UL136 and UL138 (3-7)], and resistance to NK cells [UL141 (8-10), UL142 (11, 12), and UL135 (13)]. Disabling mutations in RL13 and UL128L independently contribute to the release of high-titer cell-free virus, whereas loss of UL128L restricts virus entry to fibroblasts alone by preventing assembly of a pentameric glycoprotein complex (gH/gL/pUL128/ pUL130/pUL131A) in the virion envelope (1,14,15).The rapid selection of mutations is a major obstacle to the propagation of genetically intact "clinical" strains of HCMV, which in turn has led to significant gaps in our understanding of the affiliated disease processes (16). In particular, HCMV is largely cell-associated in vivo (17), and clinical isolates exhibit a similar phenotype in vitro (17, 18), yet most studies in the field are based on laboratory strains that produce high titers of cell-free virus (19). As a consequence, little is known about the fundamental processes involved in the infectious spread of cell-associated HCMV.In this study, we used a system tha...

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Section: -E)contrasting

confidence: 68%

Section: Discussionmentioning

confidence: 99%

Section: Significancementioning

confidence: 99%

See 1 more Smart Citation

The pentameric complex drives immunologically covert cell–cell transmission of wild-type human cytomegalovirus

Murrell

Bedford

Ladell

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

110

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“…In nonrandomized clinical studies, treatment of pregnant women with hyperimmune globulin was reported to improve pregnancy outcome in several studies (34,35,38,84) with limited effect in one study (37). As novel therapeutics, human MAbs specific for epitopes on HCMV proteins have been developed using technological advancements in human B-cell cloning (51,85,86). TBPC phenotype and differentiation capacity were rescued by anti-gB MAb treatment (Fig.…”

Section: Discussionmentioning

confidence: 99%

Human Cytomegalovirus Infection Interferes with the Maintenance and Differentiation of Trophoblast Progenitor Cells of the Human Placenta

Tabata

Petitt

Zydek

et al. 2015

J Virol

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Human cytomegalovirus (HCMV) is a major cause of birth defects that include severe neurological deficits, hearing and vision loss, and intrauterine growth restriction. Viral infection of the placenta leads to development of avascular villi, edema, and hypoxia associated with symptomatic congenital infection. Studies of primary cytotrophoblasts (CTBs) revealed that HCMV infection impedes terminal stages of differentiation and invasion by various molecular mechanisms. We recently discovered that HCMV arrests earlier stages involving development of human trophoblast progenitor cells (TBPCs), which give rise to the mature cell types of chorionic villi-syncytiotrophoblasts on the surfaces of floating villi and invasive CTBs that remodel the uterine vasculature. Here, we show that viral proteins are present in TBPCs of the chorion in cases of symptomatic congenital infection. In vitro studies revealed that HCMV replicates in continuously self-renewing TBPC lines derived from the chorion and alters expression and subcellular localization of proteins required for cell cycle progression, pluripotency, and early differentiation. In addition, treatment with a human monoclonal antibody to HCMV glycoprotein B rescues differentiation capacity, and thus, TBPCs have potential utility for evaluation of the efficacies of novel antiviral antibodies in protecting and restoring placental development. Our results suggest that HCMV replicates in TBPCs in the chorion in vivo, interfering with the earliest steps in the growth of new villi, contributing to virus transmission and impairing compensatory development. In cases of congenital infection, reduced responsiveness of the placenta to hypoxia limits the transport of substances from maternal blood and contributes to fetal growth restriction. IMPORTANCEHuman cytomegalovirus (HCMV) is a leading cause of birth defects in the United States. Congenital infection can result in permanent neurological defects, mental retardation, hearing loss, visual impairment, and pregnancy complications, including intrauterine growth restriction, preterm delivery, and stillbirth. Currently, there is neither a vaccine nor any approved treatment for congenital HCMV infection during gestation. The molecular mechanisms underlying structural deficiencies in the placenta that undermine fetal development are poorly understood. Here we report that HCMV replicates in trophoblast progenitor cells (TBPCs)-precursors of the mature placental cells, syncytiotrophoblasts and cytotrophoblasts, in chorionic villi-in clinical cases of congenital infection. Virus replication in TBPCs in vitro dysregulates key proteins required for self-renewal and differentiation and inhibits normal division and development into mature placental cells. Our findings provide insights into the underlying molecular mechanisms by which HCMV replication interferes with placental maturation and transport functions. H uman cytomegalovirus (HCMV) is the most common cause of congenital viral infection in the UnitedStates. Each year, at least 4...

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“…Studies of primary human placental cells and tissue models have identified molecular pathways that impair trophoblast differentiation [17][18][19][21][22][23][24] . Patterns of viral proteins in infected placentas suggest modulation of infection in early Since the attenuated and clinical strains exhibited markedly distinct levels of infection in placental explants, the differences were quantified by counting the number of cytotrophoblasts expressing IE1…”

Section: Under the Microscopementioning

confidence: 99%

Cytomegalovirus infection and pathogenesis in the human placenta

2015

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As shown in Figure 1, the human placenta is composed of chorionic villi bathed in maternal blood and villi that anchor the placenta in the uterine wall (decidua), attaching the fetus to the mother (panel a1). The individual chorionic villus contains a connective core with blood vessels that carry substances to the fetal circulation (panel a2). Placentation is a stepwise process whereby villus cytotrophoblasts (vCTB) attached to the basement membrane as a polarised epithelium leave the membrane to differentiate along one of two independent pathways depending on their location. In floating villi, they fuse to form a multinucleate synctiotrophoblast (STB) covering attached at one end to the tree-like fetal portion of the placenta. The rest of the villus floats in a stream of maternal blood, which optimises exchange of substances between the maternal and fetal circulation. In the pathway that gives rise to anchoring villi, cytotrophoblasts aggregate into cell columns of non-polarised mononuclear cells that attach to and then invade the uterine wall (iCTBs).

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A High-Affinity Native Human Antibody Neutralizes Human Cytomegalovirus Infection of Diverse Cell Types

Cited by 46 publications

References 89 publications

The pentameric complex drives immunologically covert cell–cell transmission of wild-type human cytomegalovirus

The pentameric complex drives immunologically covert cell–cell transmission of wild-type human cytomegalovirus

Human Cytomegalovirus Infection Interferes with the Maintenance and Differentiation of Trophoblast Progenitor Cells of the Human Placenta

Cytomegalovirus infection and pathogenesis in the human placenta

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