Multiplexed screening of natural humoral immunity identifies antibodies at fine specificity for complex and dynamic viral targets

McCutcheon, Krista; Gray, J. A.; Chen, Natalie Y; Liu, Keyi; Park, Minha; Ellsworth, Stote L.; Tripp, Ralph A.; Tompkins, S. Mark; Johnson, Scott K.; Samet, Shelly; Pereira, Lenore; Kauvar, Lawrence M.

doi:10.4161/mabs.27760

Cited by 18 publications

(34 citation statements)

References 23 publications

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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%

“…We also used M23, a mouse monoclonal antibody to UL112 and -113 (49,50), and a cocktail of mouse monoclonal antibodies directed against HCMV ICPs (MAB8121 containing clones 8B1.2, 1G5.2, and 2D4.2; Millipore, Billerica, MA). TRL345, a human IgG1 monoclonal antibody (MAb) against the conserved AD-2 epitope of HCMV gB (51), and control IgG1 MAb Synagis, which is reactive with respiratory syncytial virus (52), were provided by Trellis Bioscience, LLC (Menlo Park, CA).…”

Section: Methodsmentioning

confidence: 99%

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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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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

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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“…From 2.5 million B cells surveyed from the donor with the highest frequency of anti-AD-2 B cells, 30 MAbs were cloned by single-cell cDNA PCR and expressed in HEK293 cells. As previously described, the V H and V L germ line usage in this set of MAbs was restricted, including both published and previously unidentified gene families against this target (44). Based on its superior potency for neutralizing HCMV in vitro (44) compared to that of the other MAbs cloned in the same manner, TRL345 was chosen for further characterization.…”

Section: Screening Technologymentioning

confidence: 99%

“…As previously described, a total of 15 CDR residues were somatically mutated in TRL345, with 4 in the light chain and 11 in the heavy chain (44). None of these introduced new stability risks (such as deamidation, glycosylation, acidic cleavage, isomerization, or oxidation motifs) (66).…”

Section: Screening Technologymentioning

confidence: 99%

“…These features pose an additional obstacle to vaccine development and to quality control in HIG manufacturing. We used our previously described single-B-cell screening platform (42,43) to clone 30 different native human MAbs that bind to the highly conserved AD-2 (site I) epitope on gB (44). Although TRL345 was not the highest-affinity MAb, it had the best potency in vitro (44), substantially higher than that of either HIG or ITC-88, a well-studied MAb against this epitope (30).…”

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

Kauvar

Liu

Park

et al. 2015

Antimicrob Agents Chemother

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The History of Therapeutic Monoclonal Antibodies

Sodoyer

2016

Biosimilars of Monoclonal Antibodies

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Multiplexed screening of natural humoral immunity identifies antibodies at fine specificity for complex and dynamic viral targets

Abstract: (2014) Multiplexed screening of natural humoral immunity identifies antibodies at fine specificity for complex and dynamic viral targets, mAbs, 6:2, 460-473,

Cited by 18 publications

References 23 publications

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

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

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

The History of Therapeutic Monoclonal Antibodies

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